SWIFS Controlled Substances Procedures Manual v2.2 (08.09.2008) 149 Pages PDF

Dallas County Southwestern Institute of Forensic Sciences
CONTROLLED SUBSTANCES PROCEDURE MANUAL Version 2.2
Authorized by:
Elizabeth Todd, Ph.D., Chief Forensic Chemistry Monica Lopez, Supervisor Controlled Substances Laboratory Chris Heartsill, Acting Quality Manager August 9, 2008
Effective date:
This is a non-controlled copy of a controlled document
Dallas County Institute of Forensic Sciences Controlled Substances Procedure Manual Summary of Changes from Previous Manual Version Previous Version: Controlled Substances Procedure Manual, version 1.0 Current Version: Controlled Substances Procedure Manual, version 2.x 1. All procedure versions were changed consistent with the move to electronic format and typographical corrections were made where applicable. 2. No other substantive changes were made to the following sections: Purpose, Drug LIMS, and Testimony. 3. Evidence Management A section has been added regarding evidence security and process was clarified as needed. 4. Analysis and Reporting of Casework Previous changes by memo were incorporated into the procedure, a summary regarding analyst handling of evidence was added, and process was clarified as applicable. 5. Quality Management This section was expanded to address additional quality issues including establishment of a Quality Committee, critical reagents, housekeeping, and case review. 6. The Abbreviations and Drug and Procedure Cross Reference administrative sections are new to this manual. 7. The following procedures have been reorganized to improve utilization with minor updates as needed: Acetylated Derivatives, Acidic Drug Analysis, Alkaline Drug Analysis, Cocaine Direct Dilution, Direct Dilution, IR Procedure, LSD, Marihuana, and Weak Alkaline Drug Analysis. 8. Color Testing A quality control documentation section was added, minor updates and reorganization were performed. 9. Miscellaneous Materials More specific information regarding testing of GHB and mushrooms is provided, the section on clandestine laboratories has been moved to the Alkaline Drug Analysis procedure, and Morphine Syrup was moved to a standalone procedure and upgraded. 10. Offsite Sampling, Pharmaceutical Analysis, and Unknown Sample Screening are newly created procedures detailing current process covered in other procedures or practices. 11. Cocaine Free Base procedure was removed; it is covered in the Cocaine Direct Dilution procedure.
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory
Changes from Previous Version Version 2.0
Controlled Substances Procedure Manual Version 2.x Revisions and Corrections Effective Date 2/28/08 4/9/08 Description Authorized by Quality Management, addition of Section VI. Use of Positive and ELT Negative Controls Revision of Quality Management, Section VI. Use of Positive and ELT Negative Controls to improve instruction.
Revisions and Corrections Version 2.1

STATEMENT OF PURPOSE The Dallas County Forensic Laboratory is a division of the Dallas County Southwestern Institute of Forensic Sciences. It is an independent laboratory established through joint agreement of the Dallas County Commissioners Court and UT-Southwestern Medical School. The Laboratory is not a part of any police agency, the Dallas County District Attorneys Office, nor the Dallas County Sheriffs Office. The Laboratory operates on a fee-for-services basis, and its services are available to anyone paying the fees prescribed by the Dallas County Commissioners Court within the policies and procedures established by the Commissioners Court and the Director of the Institute. The purpose of the Drug Analysis Section of the Dallas County Institute of Forensic Sciences is as follows: 1. To analyze materials controlled by the Texas Drug Laws - including but not limited to the Texas Controlled Substances Act, Simulated Controlled Substances, Dangerous Drugs, Volatile Chemicals, Abused Glues, and Aerosol Paints (Chapters 481 - 485 of the Texas Health and Safety Code) - and the Federal Controlled Substances Act; 2. To testify in court proceedings regarding testing results, methods of analysis, and the general effects of material submitted to the laboratory; 3. To assist submitters in the identification of drug substances, raw materials, and precursors in the manufacture of these substances and adulterants and dilutants which may be present in the final product. The purpose of this manual is to outline the activities conducted by this Laboratory and to provide an overview of sample analysis strategy. Because of the unique nature of many of the samples submitted to this Laboratory, exact analytical procedures cannot be prescribed for every case. Therefore, analytical techniques described in this manual are given as guidance, and the actual choice of analytical methodologies will be left to the discretion of the Drug Analyst, Supervisor, and/or Section Chief. All variances from these procedures, however, will be described in the case record. It is the goal of this laboratory to perform analyses conforming to accepted standards of laboratory practice. OVERVIEW OF LABORATORY ACTIVITIES Samples are usually received into the laboratory by the Evidence Registrar. A unique laboratory number is assigned to each case. Evidence is stored in a sealed condition until it is opened by an analyst. During analysis evidence is stored in the evidence vault, in the personal possession of the analyst, or in the analysts personal locked storage area. The analytical process includes description, weighing, analysis, and reporting of evidence. Cases are reviewed by a supervisor, and a report sent to the submitting agency. Evidence is resealed and stored in a secure area until it is returned to the submitting agency. Testimony regarding work performed and general drug toxicology is provided as applicable.
Statement of Purpose Version 2.0
Drug Analysis Laboratory EVIDENCE MANAGEMENT: Evidence Security; Receipt and Release of Drug Evidence; Records Management I. Evidence Security A. Because of the inherent value of drug evidence and criminal penalties related to possession of controlled substances, all Drug Laboratory staff must at all times be vigilant about protecting drug substances in the custody of the Institute. B. Staff must remain actively engaged during the processing and handling of drug materials and maintain a critical perspective about their own actions, the actions of coworkers, and the actions of others in the Laboratory or at the Institute as it relates to protecting controlled substances, processing intermediates, and analytical residues. C. It is the responsibility of all staff to immediately advise a supervisor of suspected misuse or loss of suspected or known controlled substances or breaches in security or procedures that may lead to misuse or loss of suspected or known controlled substances. D. Staff actions that compromise the security of controlled substances in the possession of this Laboratory will be investigated internally and externally by law enforcement agencies as appropriate. E. Staff is subject to disciplinary action up to and including termination 1. for failure to maintain care, custody, and control of known or suspected controlled substances in their possession 2. for misuse or loss of a known or suspected controlled substances 3. for failing to advise a supervisor of known or suspected loss or misuse of known or suspected controlled substances or breaches in security or procedure that may lead to their loss or misuse 4. for failing to cooperate with investigations F. As provided for by law, staff is also subject to criminal investigation and prosecution in regard to mis-handling of controlled substances. G. Drug Laboratory Security 1. Drug Vault Routine access to the drug vault is limited by access card authorization to the Drug Evidence Registrars and the Drug Laboratory Supervisor. 2. Drug Laboratory Routine access to other Drug Lab areas is limited by access card to authorized staff only: Drug Evidence Registrars, Drug Chemists II and III, Drug Lab Supervisor, and Section Chief. 3. Only authorized staff has unescorted access to the Drug Lab areas. a) Other visitors including cleaning crews, instrument vendors, Institute staff from other labs, etc. - must be escorted at all times. (1) The person allowing entrance to a visitor is responsible for visually monitoring the activity of the visitor until the visitor is handed over to another authorized staff member or escorted out of the secure area. 4. Drug Lab areas are controlled by intrusion alarms, access cards, and key locks.
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory 1 Evidence Registration Version 2.0
a) The last person out of a Drug Lab area is responsible for key-locking the door at any time of the day. b) The last person out of a Drug Lab area at the close of business is responsible for setting the security alarm. c) All staff are responsible for using the security check list located in each Drug Lab area to monitor the status of security and evidence in the area. H. Evidence Security 1. Drug evidence should routinely be in the personal possession of authorized personnel, a locked storage locker, or vault. a) Large or bulky evidence may be left in the secure laboratory during analysis if a locked storage area is not available or practical. 2. Staff should work in a manner to ensure that evidence remains within their custody while in their possession and is not lost or contaminated. a) Trash cans should not be placed directly under a work area. 3. Personal storage areas should be kept neat and orderly; evidence should be returned to Evidence Registration in a timely manner. a) Staff must advise a supervisor if evidence remains in their personal storage area for more than one month. I. Evidence Accountability 1. Accountability: Staff is accountable for evidence, processing intermediates, analytical residues, and standards for the entire time that these items remain the responsibility of the Institute. a) Evidence may leave the Institute only by approved processes: (1) Evidence is generally released to the submitter (chain of custody required) (2) With the permission of the submitter, evidence may be released to destruction using a method approved by Institute Administration (chain of custody required) (3) Medical Examiner pharmaceuticals are released to destruction (chain of custody required) (4) Proficiency test materials, old training materials, unused standards, and other drug materials of a useable quantity must be released to destruction using a method approved by Institute Administration (chain of custody required) (5) De minimus quantities of processing intermediates and analytical residues may be disposed (a) in biohazard waste (b) as a residue in glass waste (6) Evidence may be released upon direction of the submitter or in response to court order, subpoena, or other legal direction. b) Use of drug materials, processing intermediates, or analytical residues outside standard operating procedure requires supervisory approval with transfer using chain of custody and tracking of use by a supervisor. 2. Use of evidence material for training or testing purposes:
a) With the permission of a submitting agency, drug evidence may be retained for training and testing purposes. (1) A description of retained items must be placed into the case file and signed by the analyst and a supervisor. (2) Items retained must be stored in the Drug Vault and inventoried by the Drug Lab Supervisor. II. Receipt and Release of Drug Evidence A. General Information 1. General management of evidence is described in the Evidence Handling section of the Quality Management Program. a) Additional information specific to the Drug Analysis Laboratory is described in this section. 2. Duties described in this section will usually be performed by a Drug Laboratory Evidence Registrar or the Drug Lab Supervisor but may be performed by other designated Drug Lab staff. a) The term Evidence Registrar will encompass all of the above named entities and the drug vault for the purposes of this document. 3. All Drug Laboratory staff may receive evidence in the event that an Evidence Registrar is not available. a) Staff will follow proper evidence receipt procedures and maintain evidence in a secure location until it can be released to an Evidence Registrar. b) Only designated individuals with routine evidence registration responsibilities will have access to the drug vault. 4. Responsibilities of the individual receiving the evidence include completing the chain of custody form, obtaining proper and legible submitter/billing information, assessing the status of evidence seals, and ensuring that the evidence package is properly sealed, as applicable, and stored in a secure location. a) Evidence logging may occur at a later date. B. Evidence Seals 1. To the extent feasible, evidence received by the Drug Analysis Laboratory should be properly sealed prior to receipt. a) A container is properly sealed when it is secured in a permanent manner to prevent undetected access to the contents and when its contents cannot readily escape. b) The seal must be marked with an identifying mark such as the initials of the person sealing the evidence. c) The Evidence Registrar will assess the status of the seal at the time of evidence receipt. (1) Where possible, evidence should be sealed by the submitting agency prior to receipt by the Evidence Registrar. (a) The Evidence Registrar should instruct the submitting agency to repackage and/or reseal prior to submission. (2) In any case, it is the responsibility of the Evidence Registrar to ensure that evidence is properly sealed upon receipt and to
repackage and/or seal evidence as necessary prior to storage in the vault. (a) When the Evidence Registrar must reseal submitted evidence, initials and date should be placed on the reinforced seal. 2. It is unusual that evidence received by the Drug Laboratory is of the size or nature that it cannot be properly sealed. However, under these conditions alternate methods may be used. a) Evidence should be transferred as soon as possible to the analyst for processing. b) Evidence may be stored in a hood using a unique lock/key, or if no other option is available, evidence may be placed in the drug evidence vault. c) Where possible, evidence should be properly sealed following analysis. (1) If this is not feasible, evidence should be returned to the submitting agency as soon as possible. 3. Evidence is not required to be sealed during analysis as long as it is maintained in a secure area with limited access. C. Chain of Custody 1. All evidence transfers external and internal - will be documented using the Evidence Summary form and Drug LIMS. a) Chain of custody records must be executed at the time of evidence transfer. b) For details of this procedure refer to the chapter, Drug Laboratory Information Management System, in this manual. 2. Evidence transfer must include legible documentation of the following: a) Name, signature, and affiliation of individual(s) receiving and releasing evidence (1) For Institute staff, initials or electronic identifier are acceptable in place of the signature, and affiliation is not necessary. (2) Signature only is acceptable for routine submitters whose printed name and signature are maintained in an annual signature log by the Evidence Registrar. b) Method of shipment if applicable (1) Shipping papers and/or tracking numbers, such as certified mail number, should be noted in the case file. c) Date and time of evidence transfer (time and date stamp may be used) d) General description of evidence transferred if the entire case is not transferred e) Status of specimen seal f) Case name and/or Institute case number; agency case number as applicable D. Evidence Receipt
Evidence Registration Version 2.0
1. All evidence received into the Laboratory should be accompanied by a legible transmittal sheet and/or an Institute Evidence Summary form which includes the following information as applicable: a) Submitting agency name and contact name (1) Complete address and phone are required if the submitter is not a regular client. b) Billing agency name, billing address, billing contact, and phone if billing information differs from submitter information c) Case name d) Submitting agency case number e) Officer badge number as applicable f) Analysis requested as applicable g) General description of evidence submitted 2. In-person transfer a) The submitting agent(s) will transfer custody of evidence to the Evidence Registrar by executing the chain of custody. b) The Evidence Registrar will accept evidence by completing the chain of custody. (1) At this time, the Evidence Registrar will compare the transmittal sheet or evidence submission form with the evidence for correct name, service number, and arrest date and inspect the condition of the evidence seal. (2) Questions regarding evidence and paperwork will be resolved at this time. 3. Lockbox transfer a) The Evidence Registrar may receive evidence directly from a lockbox. (1) At this time, the Evidence Registrar will compare the transmittal sheet or evidence submission form with the evidence for correct name, service number, and arrest date and inspect the condition of the evidence seal. (2) Questions regarding evidence and paperwork will be resolved as soon as possible and referred to a supervisor as needed. 4. Evidence remains in the custody of the Evidence Registrar until it is released for analysis. E. Evidence Tracking 1. Location and possession of evidence will be tracked throughout the storage and analysis process. a) For details of this procedure refer to the chapter Drug Laboratory Information Management System in this manual. F. Case Completion and Release 1. A case is complete when a Supervisor, Section Chief, or other reviewer signs the case report. a) Computerized case records should also be updated as appropriate. b) The analyst will return the signed report and the sealed evidence to the Evidence Registrar using proper chain of custody: Evidence Summary form and Drug LIMS.
c) Evidence will be stored in the Drug Evidence Vault until release. 2. Evidence is available for return to the submitter when the final report has been completed and signed. a) The Evidence Registrar is responsible for notifying a submitter that evidence is available for release. 3. To release evidence, the Evidence Registrar completes the external chain of custody on the Evidence Summary form. a) The name of the individual receiving the evidence, the date, and time of transfer are noted on the Evidence Summary form. b) If the identity of the individual receiving evidence is unknown to Drug Laboratory staff, identification must be requested and verified. c) Computerized case records should also be updated as appropriate. 4. Evidence may be released only to the submitting agency or to an entity approved by the submitting agency or in response to and in compliance with a subpoena, court order, or other legal direction. a) Evidence is usually released to the submitter; release to another entity requires supporting documentation in the case file. b) Evidence may be returned by mail or package delivery with proper notation made in the case file. 5. Controlled substances may not be released to submitters who do not have authorization to possess controlled substances. a) Disposition of these items will be arranged with the submitter on a case by case basis and may include storage by IFS, disposal, transfer to a properly designated agent, etc. 6. If the submitting agency has not arranged for disposition of evidence within 90 days of case completion, the Evidence Registrar may contact the submitter in writing requesting that the submitter collect evidence within 30 days or provide IFS approval for evidence destruction. a) If evidence remains after this 30 day period, it may be returned to the submitting agency by mail or package service, and the submitter billed the applicable shipping fee. III. Records Management A. Records management and release is described in the Control and Maintenance of Case Record Documentation and Disclosure of Information sections of the Quality Management Program. 1. Specific Drug Laboratory information is provided here. B. Primary Custodians of Records for the Drug Analysis Laboratory include the Evidence Registrar, Drug Chemist III, Laboratory Supervisor, and Section Chief. C. Drug Chemist II are Custodians of Records for case which they analyzed. D. The Evidence Registrar manages records on a daily basis and provides copies of reports as appropriate. E. The Evidence Registrar oversees transfer of case files to off-site storage. F. Copies of case reports are provided to the submitting agency and as applicable to other investigative or prosecuting entities. G. Distribution of case reports will be documented in the case file.
H. Corrections or amendments to case records will be sent to all entities receiving the original report. I. Although original case files are routinely used by the analyst in legal proceedings, it is the policy of the Institute that original records remain in the custody of Institute staff. 1. Copies of case reports and case records are provided in response to subpoenas or other legal directives and for introduction in Court proceedings. 2. Requests for release of records other than as described above will be directed to the Dallas County District Attorneys Office. J. When a case is removed from the building for example, for testimony, the analyst will make notation in the case file.

STORAGE, TESTING, AND DESTRUCTION OF MEDICAL EXAMINER PHARMACEUTICALS AND DRUGS Pharmaceutical and drug evidence collected or received by the Dallas County Office of the Medical Examiner (OME) is submitted to Drug Evidence Registration for storage and destruction. In some cases, the Controlled Substances Laboratory will perform testing on this evidence at the request of Medical Examiner staff. Additional information regarding the submission of drug evidence may be found in the Evidence Registration section of the Controlled Substances Procedure Manual. I. Submission of Medical Examiner Drug Evidence to Drug Evidence Registration A. Submission Using the Field Agent Lockbox Field Agents will deposit properly sealed evidence in the Field Agent Drug Lockbox. Evidence must be submitted in tamper evident plastic bags or other containers with the Field Agents initials across any non-factory seals. A case barcode will be placed on the evidence bag. If more than one bag is submitted, the Field Agent will write the case number and case name on each additional bag. A Medical Examiner Drug Submission form must be completed and submitted with each case submitted. Each case submission to the Field Agent Drug Lockbox must be documented on the Medication Drop Log.
B.
Submission Directly to the Drug Lab Evidence which requires testing is often submitted directly to Drug Evidence Registration by the Office of the Medical Examiner. In accepting Medical Examiner evidence for testing, the Drug Evidence Registrar accepts evidence on behalf of both the Medical Examiners Office (as it relates to this current policy of storage and destruction) and the Drug Laboratory (as it relates to testing). A Medical Examiner or Field Agent may contact a Drug Evidence Registrar to request analysis of evidence previously submitted.
II.
Management of Medical Examiner Drug Evidence A. B. C. The Drug Evidence Registrar ensures that all Medical Examiner drug evidence has a proper evidence submission form and is properly sealed. The Registrar enters evidence into the Medical Examiner drug evidence tracking system and store evidence in a secure location. The Registrar may release drug evidence to a Field Agent upon request with completion of appropriate chain of custody. Evidence may be returned to the Drug Evidence Registrar in person or dropped into the Field Agent Drug Lockbox by the Field Agent returning the drug evidence. The Registrar will complete the
1 ME Pharmaceutical & Drug Evidence Version 2.0

chain of custody as appropriate. It is expected that the Field Agent will return drug evidence in a timely fashion; variance should be brought to the attention of a Supervisor. Release of evidence to other than a Field Agent or Medical Examiner requires written permission of the Chief/Deputy Chief Field Agent or Chief/Deputy Chief Medical Examiner. Chain of custody will be documented in the appropriate case file.
D.
III.
Retention of Medical Examiner Drug Evidence after Case Completion A. B. C. Medical Examiner Drug Evidence will be held for at least 90 days post collection. Evidence will be considered for destruction upon case completion or 90 days post collection whichever is later. Requests to retain drug evidence beyond the period noted above must be submitted in writing to the Chief of Forensic Chemistry by the individual requesting evidence retention and conform to the requirements noted in the Medical Examiner Drug Evidence Disposal and Hold Policy (refer to Appendix). On their own behalf, Institute staff may request retention of Medical Examiner drug evidence beyond the routine retention period by sending a written request to the Chief of Forensic Chemistry identifying the following: Case Number Decedent Requestor Name Reason for Hold Length of Hold Date of Hold Request
D.
III.
Disposal of Medical Examiner Drug Evidence A. Medical Examiner Property 1. As needed, Medical Examiner drug evidence which exceeds 90 days from collection, is not held for Medical Examiner evidentiary purposes, or held per the Medical Examiner Drug Evidence Disposal and Hold Policy will be disposed. The Drug Evidence Registrar will prepare a list of cases for destruction. Cases designated for disposal will be placed into a disposal container, sealed, witnessed by two IFS staff, one of which should be a Drug Evidence Registrar. Cases included in each disposal box will be noted, the box will be sealed, and the seal initialed by both staff. The box is deemed ready for destruction provided the seal remains intact. This process must be repeated if the seal is broken for example to add or remove a case.
2. 3.
ME Pharmaceutical & Drug Evidence Version 2.0

B. Forensic Laboratory Property 1. Certain drug materials collected by the Institute Forensic Laboratory including but not limited to drug materials remaining from analytical procedures, drug standards, and drug materials not retrieved by submitting agencies will be identified for disposal. These materials will be placed into a disposal container and the contents witnessed by two Institute staff, one of which should be a Drug Evidence Registrar. Contents of the box will be noted, the box sealed, and the seal initialed by both staff. This process must be repeated if the seal is broken.
2.
C.
Order for Destruction An Order for Destruction (refer to Appendix) listing the cases and/or drug materials to be disposed will be submitted to the Chief Medical Examiner, who presides over the Court of Inquest, for signature and authorization to dispose.
D.
Disposal Regulations and Policies 1. Possession and disposal of drug substances are regulated by the Drug Enforcement Agency (DEA), the Texas Commission of Environmental Quality (TCEQ), and the Environmental Protection Agency (EPA). Disposal facilities may also have policies and procedures which must be followed. An appropriate disposal facility will be selected by Institute Administration and Purchasing. Specific arrangements must be made with the disposal facility prior to each destruction. Only finished drug substances (including plant material) may be sent to the disposal facility. Specific arrangements must be made for disposal of clandestine laboratory chemicals, aerosol cans, or other types of waste. Institute staff or other entity designated by the Chief Medical Examiner must retain custody of drug evidence until actual physical destruction at the disposal facility; this is called a witnessed burn. A certified peace officer or DEA licensee must also accompany Institute evidence to the disposal facility. Disposal is documented on a manifest and/or other certificate of destruction. The destruction records must be signed by two individuals on behalf of the Institute.
2. 3. 4.
5.
6. 7.
IV.
Retention of Records The following records will be retained on the same schedule as other Medical Examiner records: the original Order of Destruction, inventory of cases/materials disposed, original manifest or certificates of destruction, Medical Examiner Drug Submission forms, and other relevant materials related to the destruction.

SOUTHWESTERN
INSTITUTE OF FORENSIC SCIENCES

AT DALLAS
5230 Medical Center Drive Dallas, Texas 75235
TELEPHONE: 214-920-5990 FAX: 214-920-5812
MEDICAL EXAMINER DRUG EVIDENCE DISPOSAL AND HOLD POLICY Medical Examiner drug evidence is authorized for disposal upon case completion or 90 days post collection whichever is later unless an interested party acts as described below: Storage of Medical Examiner drug evidence beyond the standard storage period noted above requires specific action on the part of the agency/individual requesting that specimens be held. Medical Examiner drug evidence may be held for one year increments provided all of the following conditions are met prior to 90 days from sample receipt by the Instiute in year one or prior to the anniversary date of evidence receipt by the Institute for evidence held per this policy beyond one year): A. The Chief of Forensic Chemistry must receive a written request from the agency/individual requesting that drug evidence is retained for one or one additional year. This written notice must include the case name; the Medical Examiner case number; the name, phone number, and complete mailing address of the agency/individual requesting specimen hold; and a description of the materials to hold, AND The Chief must be in receipt of a check for the amount of the Annual Specimen Storage Fee authorized by the Dallas County Commissioners Court. The check must be made payable to Dallas County and reference the case name and/or case number, AND It is the responsibility of the agency/individual requesting evidence hold to renew the annual evidence hold prior to the anniversary date of evidence receipt at the Institute. Otherwise samples will be disposed per Institute policy. The Institute will not provide annual notice of this requirement to renew the sample hold request and provide payment, AND Dallas County reserves the right to change this policy at any time. Reasonable effort will be made to notify the agency/individual requesting the evidence hold; however, no guarantee is made in this regard. It is the responsibility of the agency/individual requesting evidence hold to keep the Institute advised of address changes, etc.
B.
C.
D.
All correspondence and payment to Dallas County must be sent to the Chief of Forensic Chemistry, Dallas County Institute of Forensic Sciences, 5230 Medical Center Drive, Dallas, TX 75235. All correspondence and payment must be received prior to the routine destruction date or anniversary date of evidence receipt at the Institute. This policy pertains to retention of drug evidence only and does not guarantee evidence release. Release of evidence requires approval of the Chief Medical Examiner, submitting agency, and/or proper legal authority. In addition, the entity or individual requesting release
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must provide proper authorization to possess drug materials as applicable, for example a DEA number. Additional fees related to evidence release are the responsibility of the requestor.
Drug Analysis Laboratory OVERVIEW OF ANALYSIS AND REPORTING OF CASEWORK The following provides guidance regarding analysis and reporting of suspected drug evidence. Analysts should consult with supervisors as needed in the implementation of this guidance. Variance from written procedure must be documented in the case and reviewed and approved by a supervisor. I. Internal Chain of Custody A. The internal chain of custody begins when the analyst takes possession of cases from the Evidence Registrar. B. All internal transfers of evidence are documented on the internal chain of custody section of the Evidence Summary form which is the official chain of custody record. 1. Internal transfers will also be recorded as applicable in Drug LIMS to facilitate administrative tracking of evidence. C. When evidence is transferred from one analyst to another, the evidence may be sealed or transferred in person unsealed. 1. If evidence is transferred in person unsealed, both analysts must verify the inventory at the time of transfer and document this by initialing and dating the inventory log. II. Sealing of Evidence A. Evidence remains sealed until it is processed, i.e. inventoried, weighed, sampled, etc. 1. Only one case is processed at a time. 2. Evidence is resealed at completion of processing. B. Immediately prior to sealing the evidence container, the analyst will verify the presence of all items listed on the inventory which are not otherwise accounted for in the case file, for example materials used in analysis. 1. All exhibits will be placed into the evidence bag with any additional residue tubes if applicable. 2. Any additional items included will be marked added by lab, lab bag, etc. 3. The analyst will document concurrence with the inventory by writing sealed, initials, and date in the upper left portion of the transmittal sheet. C. All evidence will be sealed in a secure, tamper evident manner. 1. The seal must be initialed and dated by the analyst sealing the evidence. 2. In the specific case of a heat sealed evidence bag, the analyst will initial and date the inside of the bag along the cut area and heat seal the bag through the initial and date. D. The internal chain of custody form will be completed as the sealed evidence is released from the custody of the analyst to the Evidence Registrar; this transfer will also be documented for administrative purposes using the evidence transfer function of Drug LIMS. E. If evidence is reopened at another time, the inventory will be verified against the original inventory taking into consideration materials accounted for otherwise in the case file. 1. Concurrence with the inventory will be documented by writing reopened, initials, and date in the upper left portion of the transmittal sheet.
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory Analysis and Reporting of Casework Version 2.0
2. Immediately prior to resealing the evidence container, the analyst will verify the presence of all items listed on the inventory which are not otherwise accounted for in the case file. The analyst will document concurrence with the inventory by writing sealed, initials, and date in the upper left portion of the transmittal sheet. F. Exit Weight of Primary Containers 1. The sealed evidence container will be weighed at the time the container is sealed and the weight entered in the case file as exit weight. 2. If evidence is reopened, the process will be repeated with the new entrance weight and exit weight noted in the case file. 3. It is the responsibility of the analyst and case reviewer to include evaluation of entrance and exit weights as a part of routine case review. III. Inspection and Weighing of Primary Evidence Containers A. The analyst will inspect the evidence container to determine if it is sealed. 1. If the evidence container is sealed, the analyst will indicate this by writing sealed and/or initialing on the evidence container. 2. The analyst will also note that the evidence was sealed by indicating sealed on the internal chain of custody. B. The analyst will weigh the sealed primary evidence container(s) to determine the submission weight. 1. The weight is entered on the Evidence Summary form. 2. The balance identification number is also recorded. 3. In the case of multiple primary evidence containers, each container will be weighed separately. 4. Primary container weights will be compared to the agency submission weight. a) Significant variation will be witnessed by a second chemist and immediately brought to the attention of the supervisor who will determine whether to suspend analysis pending notification of the submitter. C. The analyst will open the evidence. 1. To open evidence in a heat sealed bag, the analyst will cut the factory seal (if present), write the case number and initials on the cut strip, and place the strip into the evidence bag. 2. As possible, evidence will be opened in a manner which leaves the original seals present. D. The analyst will then inventory the evidence. IV. Inventory and Marking of Evidence A. The analyst is responsible for making an inventory of all submitted evidence. 1. Upon opening the primary evidence container, the analyst will document the contents of the evidence on the Evidence Summary form. 2. The analyst will compare the submitting agencys inventory to the contents of the opened evidence. a) Any variation in the agencys inventory, such as addition or deletion of suspected drug evidence and item count discrepancies, will be immediately witnessed and verified by another analyst who will also initial and date the inventory description.
(1) The primary analyst will also immediately notify a supervisor who will determine if the submitting agency should be notified. B. Identifying Evidence 1. Each case is assigned a unique case number by the Evidence Registrar. 2. The analyst will usually determine exhibit numbers. a) Like populations within a case will be grouped into exhibits. Like populations are pieces of evidence of similar size, packaging, and/or general appearance. b) Evidence will be identified on worksheets and on the report by the case number and an exhibit number. c) The exhibit number will be a combination of letters and numbers as follows: (1) The primary evidence containers will be identified as Exhibit 1, 2, 3, etc. (Ex. 1). (2) Like populations of evidence inside the outer evidence container will be grouped together and identified by a number and letter. (a) For example, Ex. 1A will include all similar evidence found inside the outer container. (b) Ex. 1B will include a second type of evidence found inside the outer container. (c) Loose substance found inside the evidence bag will generally be considered one exhibit and identified by a number and letter, for example, Ex. 1C. d) If there is more than one item in each population described above, the exhibits actually analyzed will be further identified by a number. (1) For example, Ex. 1A1 and Ex. 1A2 represent two pieces of evidence from evidence group 1A which were analyzed. e) Where possible, the submitting agencys item numbers will be used if they fit the above format. If the submitting agencys item numbers are not consistent with the above scheme, then the analyst will usually identify evidence using the above scheme and will note the submitting agencys corresponding evidence numbers in parenthesis on the transmittal sheet and on the report, for example, Ex. 1A1 (Item 4). 3. Each piece of evidence will be labeled with the unique case number. a) If the evidence cannot be labeled, the proximal container must be marked with the case number. 4. Pieces of evidence which are analyzed should be marked with the case number, exhibit number, and analysts initials. a) It must be clear from the case record or evidence labeling which evidence received what testing. b) If the evidence cannot be marked, the information must be placed on the proximal container. 5. If the proximal container was added by the laboratory, it should be so marked and noted. V. Analytical Protocols A. Analytical Protocol Forms
1. Drug Identification Protocol and Marijuana Protocol forms are used to summarize all analytical testing performed for a case. a) The analyst enters the starting date for the case, the case number, exhibit number, and a description of the evidence. b) A record of the testing is entered on the form as analyses are performed; transcription of analyst notes from scratch paper is not acceptable. c) Calculations are also performed directly on this form. d) In general, each exhibit analyzed should have a Drug Identification Protocol form. (1) Color testing or tentative identification of additional exhibits not otherwise analyzed may be summarized on a separate Drug Identification Protocol form or other summary sheet. (2) Calculations must be shown in the case file. B. Weighing of Evidence 1. A total weight of the suspected controlled substance, less packaging, must be obtained for each population of evidence analyzed with the exception of a residue. a) Weights should be entered onto the Drug Identification Protocol or Marihuana Analysis Protocol for each applicable exhibit. b) The analyst must verify that the balance to be used is properly calibrated. c) The balance identification number used for each weight is recorded on the protocol form. d) The total weight of the exhibit(s) is usually determined directly by measuring the weight of the material. (1) In some cases, it may be necessary to determine the exhibit weight by difference (total weight of packaging and contents minus the weight of packaging). In this situation, the weight of packaging is determined by actual weight or by averaging the weight of packaging from several representative items within one exhibit. 2. For non-residue samples, an aliquot of each exhibit chosen for quantitative analysis will be weighed and placed into a screw cap vial or other suitable container for further quantitative analysis. 3. Reporting weights. a) Weights from analytical balances are usually reported in milligrams. b) Weight determined from a top loading balance is usually reported in grams. 4. Suspected marihuana is usually determined in ounces and pounds. C. Analytical Testing 1. Selection of Evidence for Analysis a) If numerous pieces of evidence are submitted to the Laboratory, the analyst will select evidence for analysis based on the following criteria: (1) Item(s) specifically requested by submitting agency (2) Chemical(s) in the highest penalty group (3) Evidence with the greatest weight (4) Other as determined by the analyst and/or supervisor
b) If initial testing is negative, testing will continue until a controlled substance is identified or until all exhibits have tested negative. c) In general, a complete analysis will be performed on at least one piece of evidence in a case when a controlled substance is identified. (1) A complete analysis usually consists of color test(s) and/or preliminary identification of marked pharmaceutical preparations, unique qualitative analysis (such as GC/MS or IR), and quantitative analysis (such as GC). (2) Additional testing may be performed as noted above, as determined by the analyst and/or supervisor, or when weight will increase the penalty group. (3) Any combination of analytical tests may be performed on additional pieces of evidence. d) Evidence may also be composited for testing. (1) If evidence is to be composited, each individual piece of evidence to be combined will be color tested or otherwise tested to verify the presence of a controlled substance. (2) To preserve the physical condition of the evidence as submitted and to minimize health hazards related to handling evidence, a representative sample from each piece of evidence to be combined will usually be taken, mixed into a homogenous sample, and used for analysis. (a) In some cases, entire exhibits may be combined. (b) It must be clear from the Protocol form whether a single exhibit or composited exhibits were analyzed and whether a portion of each item or the entirety of each item was composited. e) If the number of items or exhibits needed to reach the applicable aggregate weight limit is large, analysis of a valid subset of the entire population of samples may be performed using color testing, compositing, and analysis usually by GC/MS or IR. Quantitation may also be performed. f) Documentation of materials analyzed and analyses performed must be clearly documented in the case file. g) If there are numerous exhibits in a case and a substantial portion of exhibits will be processed for analysis in a manner which changes the physical characteristics of the evidence, the analyst must photograph the evidence prior to analysis to establish a record of the original appearance of the evidence as submitted. 2. Color Testing and Preliminary Identification of Marked Pharmaceuticals a) Color test(s) will usually be performed on a sample of each exhibit to be analyzed using the methods described in the Color Testing procedure. (1) If a color change occurs, the color of the spot test(s) performed and the preliminary identification should be recorded on the Drug Identification Protocol and/or Marihuana Analysis Protocol forms.
(2) If there is no change in color, a negative result must be noted. b) Pharmaceuticals may be tentatively identified by markings using the PDR and other indexes. This process should be documented on the Protocol form or other summary sheet. 3. Qualitative Identification a) Unique identification of a controlled substance is typically made using GC/MS or IR. Analytical methods are selected and noted on the Drug Identification Protocol or Marihuana Analysis Protocol. (1) The mass spectrum of any substance named in the final report must be included in the case file with a spectrum of the appropriate standard. (a) A copy of the total ion chromatogram should also be included, and the identity of peaks reported on the final report must be labeled on the total ion chromatogram. (b) In the case of a residue, a blank using the same analytical program will be run immediately prior to the analysis of the unknown and included in the case file. (2) The IR spectrum of the specimen analyzed must be included in the case file along with a spectrum of the appropriate standard. 4. Quantitation a) Quantitation is usually performed by GC; although other techniques such as UV may be used. The quantitative method used is noted on the Protocol sheet. b) Written quantitative procedures may be found in the Procedure Manual for the Drug Analysis Laboratory. c) For GC analysis, the peaks in the chromatogram that correspond to the controlled substance shall be clearly labeled. (1) In the case of a residue, a blank analysis using the same analytical program will be run immediately prior to the analysis of the unknown substance and included in the case file. d) The quantitative procedure including a summary of the extraction procedure for each analytical method must be noted on the Drug Identification Protocol Sheet. (1) All calculations related to quantitative analysis are performed on the protocol sheet including dilutions made. (2) Calculations are made using all figures reported from the analytical balance. e) Instrument output is included as a part of the case file. 5. Identification of Non-controlled Substances and Ancillary Pharmaceuticals a) If no controlled substance is identified in the analytical process, this will be stated on the report. b) Marked dangerous drugs, over the counter preparations, and other preparations may be identified through visual inspection using manufacturers markings, size, color, etc. (1) The reference used to visually identify a substance should be noted in the case file.
(2) When these methods are used, it must be clearly stated on the report; for example, The material was visually identified as ___ stated by the manufacturer to contain ____. 6. Use of Non-standard Methods a) Because of the unique nature of many forensic specimens, exact analytical procedures cannot be prescribed for every case. Therefore, analytical techniques described in this manual are given as guidance, and the actual choice of analytical methodologies will be left to the discretion of the Drug Analyst, Supervisor, and/or Section Chief. (1) All deviations from written procedures must be documented in the case file. (2) Use of methods not included in this manual requires approval of a supervisor and must be documented fully in the case file. 7. Verification of Case Number and Use of Bar Codes a) Throughout the analytical process, containers must be properly labeled with a unique case/exhibit number. b) Analysts must verify appropriate sample number prior to each specimen transfer. c) Bar codes are used where possible to verify sample identity. (1) If the barcode is not properly read by the reader, the vial number should be verified and noted by a second chemist. 8. Calculating and Reporting Results a) Results to be included on the final report are entered in a clear manner on the Drug Identification Protocol; this includes the name of substances identified, the amount of any substances quantitated, and a percent of controlled substance in the exhibit tested if applicable. (1) To calculate concentration of a controlled substance, all figures recorded from the balance and the GC will be used. (2) Total exhibit weight measured on an analytical balance will be recorded to 1/10 mg or as consistent with the accuracy of the balance. If a top loader is used, weight will usually be recorded to 1/100 g or as consistent with the accuracy of the balance. (3) In each exhibit, the calculated amount of controlled substance will usually be truncated and reported as follows: Drug Analysis: Calculated Result Reporting Format 0 - 1 mg 0.x mg 1 - 99 mg xx mg 100 - 999 mg 0.xx grams 1 - 1.49 grams 1.xx grams 1.5 29.9 grams xx.x grams 30 - 999 grams xxx grams 1000 grams and up 1.xx kilograms LSD is usually truncated to a whole number (x micrograms).
Marihuana Analysis: Sample Weight 0 - 0.009 ounces 0.01 - 0.99 ounces 1.0 - 15.9 ounces 1.00 - 9.99 pounds 10 pounds and higher Reporting Format < 0.01 ounces 0.xx ounces xx.x ounces x.xx pounds xx.x pounds
(4) Percentage of the controlled substance in the exhibit is often useful to users of the report. When a percentage is to be included in the final report, it will usually be calculated by dividing the reported amount of drug by the reported amount of material times 100. Percentages will usually be rounded and reported as follows: Calculated Result <1% and incomplete GC/MS < 1% and complete GC/MS 1 - 9.9% 10% and greater Reporting Format quantity not reported insufficient for quantitation or < 1% x.x% xx%
(5) Total weight of the case will usually be calculated by adding the reported weights of each exhibit and truncating to two decimals. b) The Drug Identification Protocol and Marihuana Analysis Protocol are completed by entering the analysts initials and the completion date. VI. Reporting A. The final report must accurately reflect the work performed and be fully supported by data present in the case file. B. The analyst will review the results of analytical testing to determine whether or not a controlled substance is present and how much material is present. C. The analyst will then write the conclusions in a report using the format on the Controlled Substances Laboratory Report or Marihuana Results of Examination forms, sign, and date the form. 1. The naming of chemicals in the report will be consistent with those used in the Texas Controlled Substance Act as applicable. 2. The only abbreviations used on reports are mg for milligram(s) and mL for milliliter(s); all other units are spelled out. D. If two or more exhibits are found to contain the same controlled substance, it may be valuable to the user of the report to include a sentence stating the total weight of the multiple exhibits. 1. In this case, the reported weights and amounts from appropriate exhibits are added to determine the total weight of controlled substance including adulterants or dilutants. E. The typewritten report will include the following information where applicable:
VII.
1. administrative information: unique forensic laboratory case number, submitter case or reference number, submitter address, case name, arresting officer badge number, evidence receipt date, submitting individual information, receiving individual information, general description of the item received, and the total weight of the primary container(s) 2. analytical information and conclusions: a listing of each exhibit analyzed, description of exhibit(s) analyzed, identification of any controlled substances identified, adulterants or dilutants as applicable, weight of the controlled substance, purity of the controlled substance, and the total weight of the exhibit. 3. a list of exhibits suspected of being a controlled substance which were not analyzed 4. a list of analyses performed 5. signature of the analyst and reviewer F. The analyst will perform a review of the typewritten report and ensure that it accurately reflects the results of analytical testing, is free of typographical errors, and is consistent with laboratory policy. G. The analyst will sign the report and complete a billing form. H. The case file is then sent to a supervisor for technical/peer review and countersignature. 1. In an emergency situation in which a supervisor is not available, the case may be reviewed by a second analyst who countersigns the case reviewed by .... 2. The technical/peer reviewer will initial each page of the examination documentation. I. A copy of the report will be provided to the submitting agency and to the courts by request. J. A notation will be made in the case file of the mailing/faxing date and entities receiving a copy of the completed report. 1. By standard practice, the Dallas County District Attorneys Office has access to electronic unsigned reports for the Dallas Police Department; this is not noted in the case file. K. The case file consists of the typewritten report, handwritten reports or notes, the Drug Identification Protocol or Marihuana Analysis Protocol, all analytical work done by the analyst which is reflected in or relied upon for the written report, instrument printouts, Description of Evidence form, the transmittal sheet, record of conversations, photos, peer review sheets, and/or other relevant information. L. Corrected, supplemental, and amended reports 1. If it is determined that a revised or corrected report must be generated, the report will bear the notation Corrected Report. Copies of the corrected reports will be sent to agencies receiving the previous copy of the report. The corrected report will be filed along with the original case in the case folder. 2. If additional work is requested on a completed case, the report of additional testing will be marked Supplemental Report and sent to all agencies receiving the previous report. 3. If a new report is written by reporting work previously done and the additional requested work, the report will be marked Amended Report. Security of Evidence and Reports A. Evidence is stored sealed as received from the submitting agency in the evidence storage
Analysis and Reporting of Casework Version 2.0
VIII.
B. During analysis evidence is stored in the vault, in the personal possession of the analyst, or in the analysts personal locked storage area. C. After analysis, evidence is sealed and stored in the evidence storage vault until its release/return to the submitting agency. D. The completed case file, consisting of all records contributing to the reported results, is maintained by the Evidence Registrar. 1. Case files may be stored on site or off-site in the Dallas County Records Storage facility. 2. Retention of files is in accordance with Dallas County policy as implemented by the Dallas County Records Manager. Integrity of Evidence A. The quality of forensic analyses depends on preserving the integrity of the evidence. The analyst must routinely use critical judgement and good lab practices and techniques at all times including but not limited to the following: 1. One case is processed at a time; although samples may be extracted in batches. 2. Case numbers or bar code labels used on test tubes and other containers are cross checked with the case and exhibit number. 3. When bar codes are read by analytical equipment, the bar code must match with the expected barcode number or vial identity verified by a second chemist. 4. A fresh weighing paper or weighing boat is used during sample weighing; balance calibration is verified prior to use. 5. New, unused test tubes are used for laboratory analysis. 6. Non-disposable equipment such as spatulas are thoroughly cleaned between use. 7. Case numbers and exhibit numbers on instrument records are compared with sample numbers. 8. The case number and handwritten initials of the record generator must appear on each page of a case file. 9. When corrections are necessary to case records, the part in error will receive a single strike through which is initialed by the analyst or responsible individual, and the correction will be written nearby. Correction fluid will not be used. 10. During analysis, the evidence must remain under the personal control of the analyst unless it is formally released to another analyst for further testing. In an emergency, supervisors may access evidence. 11. Gloves should be changed as needed.
IX. Summary of Analyst Evidence Handling Process A. Initial Opening and Closing of Evidence 1. Obtain the submission weight of entire evidence package and enter on Evidence Summary sheet. 2. Open evidence where possible without destroying other seals. Initial and date any strip cut from the evidence bag and place into the evidence bag. 3. Perform a complete inventory of all evidence. Initial and date the Evidence Summary sheet under the inventory. 4. Compare Lab inventory to agency submittal inventory. If there is variance in drug evidence, advise another analyst to immediately verify the inventory and
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sign and date the Evidence Summary sheet. Advise a supervisor immediately.** 5. Process evidence. 6. Re-inventory evidence* as it is placed into the evidence bag and reseal evidence using a proper seal. 7. If the reinventory is reconciled, write sealed with initials and date in the upper left of the Evidence Summary sheet. If the inventory does not match, advise a supervisor immediately.** 8. Obtain the exit weight of the entire evidence package and enter on Evidence Summary sheet. B. Reopening evidence 1. Obtain a new submission weight of the entire evidence package and enter on Evidence Summary sheet. 2. Open evidence where possible without destroying other seals. Initial and date any strip cut from the evidence bag and place into the evidence bag. 3. Reinventory evidence*. 4. If the reinventory is reconciled, write reopened, date and initials in the upper left of the Evidence Summary sheet. If the inventory does not match, advise a supervisor immediately.** 5. Perform activities. 6. Reinventory evidence* as it is placed into the evidence bag and reseal evidence using a proper seal. 7. If the reinventory is reconciled, write sealed, date and initials in the upper left of the Evidence Summary sheet. If the inventory does not match, advise a supervisor immediately.** 8. Obtain a new exit weight of the entire evidence package and enter on the Evidence Summary sheet. C. Related Issues 1. Evidence remains sealed until it is opened for processing. 2. Only one case is opened and processed at a time. 3. Evidence is resealed at the conclusion of processing. 4. All seals will meet the description of proper sealed detailed in the Quality Manual. 5. It is the responsibility of the analyst and the case reviewer to include evaluation of submission and exit weights as part of routine case review. 6. If room is not available on the Evidence Summary sheet, use the back of the Evidence Summary sheet or add an additional sheet of paper. All entries should be clearly identified as to their purpose. D. Notes 1. *Reinventory evidence - To verify the presence of all items listed on the inventory which are not otherwise accounted for in the case file, for example those used in analysis. 2. **Supervisory responsibility Document and investigate the situation in a timely manner. Advise Section Chief. Advise submitter as applicable. Initiate Request for Review as applicable.
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Analysis and Reporting of Casework Version 2.0
DRUG LABORATORY INFORMATION MANAGEMENT SYSTEM (LIMS) I. General Information
The Drug LIMS is designed to provide evidence tracking administrative internal and external chain of custody, result entry and report writing, billing entry and reporting, and storage and transfer of non-official report copies to the Dallas County District Attorneys office. II. User Log-In
The User is defined as the person who is entering data into the Drug LIMS. Each chemist and evidence custodian has an individual log-in name and password for security purposes and ease of system use. When logging in to the system, double-click the Drug LIMS icon found on the computer desktop. The user should enter their initials in the User box or pick their initials from the drop down list. The user should then enter their unique password in the Password box and hit <Enter> or click on Log In. This will allow the user entry into the Drug LIMS and the Main Menu. III. Menu Structure
Main Menu Evidence Transfer Case Submission Assign Case to Chemist Transfer of Case Release Evidence Print Chain of Custody Exit to Main Menu Results Results Entry Case & Billing Reports Review Cases Exit to Main Menu Queries Find Case Exit to Main Menu Utilities Change Password Clear Case Transfer Table Exit to Main Menu Log Out
Laboratory Information System Version 2.0
IV.
Evidence Transfer A. Case Submission
This function is used for the purpose of entering new cases from submitting agencies into Drug LIMS and initiating chain of custody. Case and agency information may be input using two different strategies: manual entry and download of agency data. Manual Entry: 1. Input all agency information in text fields provided (Agency, Date, Received From) 2. Input all case information for each case submitted, as applicable (DefendantName, DOA, TAG#, SER#, Invc#, Status, Priority) 3. Assign FL# by either typing the number in the FL# field, by barcoding the number into the field, or by using the Assign FL#s button. Utilizing the Assign FL#s assigns the first case in the field with a User inputted number, and assigns the following cases numbers by increments of one. 4. Data is updated and cases are ready for assignment once the Accept Data button has been clicked. (NOTE: The Evidence Registrar may exit from the Case Submission form without assigning FL numbers. The information is saved in the data table until such time as the FL numbers are assigned and accepted.) Downloaded Entry from Disc: 1. Insert the floppy disk into the disk drive. 2. Click the Import Cases from Floppy Disk button. 3. Verify imported information. 4. Assign FL numbers as described above in steps 3 and 4 (Manual Entry). Note: A Supervisor must input billing and mailing data for new submitting agencies before evidence can be logged in. B. Assign Case to Chemist
This function is used for the purpose of transferring cases from the sealed evidence vault/evidence registrar to a chemist for analysis. 1. From the Evidence Transfer menu, select Assign Case to Chemist button. 2. The receiving chemist must then log into the Drug LIMS system by entering their User ID into the To field, entering their password into the To Password field and tabbing to the Cases to transfer or hitting <Enter>. (Default information for the currently logged on user, i.e. the user transferring the evidence, and the date are automatically filled in.) 3. The evidence registrar will then initiate the Internal Chain of Custody Form in hand written notation, and electronically by either barcode scanning the transferred case or by manually picking the case out of the generated pick list, clicking the Transfer check box next to the case number. (NOTE: Rush cases appear at the top of the pick list, followed by Expedite, and Normal cases.)
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory 2 Laboratory Information System Version 2.0
4. The transfer is completed when the chemist receives the cases, completes the Internal Chain of Custody Form, and the Accept button is clicked updating the tables. The chemist may then receive a copy of the cases that they have received by printing the generated spreadsheet. C. Transfer of Case
This function is used to transfer a case from one chemist to another chemist or from a chemist back into the evidence vault or evidence custodian. 1. The chemist who is making the transfer must log into the Drug LIMS system. From the Evidence Transfer menu, select Transfer of Case button. All cases in the chemists possession will appear on the screen. (Default information of the user transferring the evidence and the date are automatically filled in). 2. Select the purpose of the transfer: Return to Vault or Transfer to Chemist. 3. The receiving chemist or evidence custodian must then log into the system as described above in step 2 of Assign Case to Chemist. 4. The internal chain of custody is completed by written notation on the Internal Chain of Custody Form, and electronically by either barcode scanning the transferred case or by manually choosing the case from the generated pick list, and clicking the Transfer check box next to the case number. 5. The transfer is complete and the tables are updated when the Accept button is clicked. D. Release Evidence
This function is used to transfer evidence from the Drug Laboratory evidence vault back to the submitting agency once analysis has been completed. 1. From the Evidence Transfer menu, select Release Evidence button. 2. Select the agency from the pick list, or by typing in the abbreviated agency code. All completed cases for that agency are listed in the table. (Default information of the current logged-in user transferring the evidence and the date are automatically filled in). 3. Fill in the Released to field. (If releasing evidence to an agency other than the submitting agency, or if a special note is needed in the external chain of custody indicate this information in this field.) 4. The evidence registrar will then complete the external chain of custody in hand written notation and have the receiving agency representative sign the transmittal sheet. The electronic chain of custody is completed by either barcode scanning the transferred case or by manually choosing the case from the generated pick list, and clicking the Transfer check box next to the case number. 5. Select the Accept button and a hard copy of the released cases is printed to the network printer.
V.
Results
This menu item is used to enter evidence descriptions, type of evidence submitted, results and weights of examinations, examinations performed, billing information, and any other pertinent information found on the case report. To enter the results of an examination on a particular case, choose the Results button on the Main Menu, which will open a new Results Menu. A. 1. 2. 3. 4. Results Entry (Drug Cases)
The user completing the case must be logged into Drug LIMS. From the Results menu, select Results Entry. Scan the bar code or manually type in the desired FL#. Enter the number, description of the evidence submitted, and weight and units of the evidence bag(s), box(es), etc. The drop down lists may be used for this operation. (i.e. One heat sealed evidence bag weighing 28.3 grams). Refer to reporting results section in Analysis and Reporting of Casework. 5. Enter the exhibit number. 6. The Analysts initials will appear in the Analyst box. The default is the User currently logged into Drug LIMS. 7. The Requesting Agency is defaulted from the evidence registration process, but should be verified before proceeding. 8. Enter the number of items tested. NOTE: entering one will create the item container in singular form, any other entry will result in a plural format. 9. Enter an item description and container. The drop down lists may be used for this operation. 10. Enter the description of the evidence exhibit and the substance. The drop down lists may be used for this operation. NOTE: A comma will be placed between the two descriptives; if only one adjective is needed, use only the first description box. If additional descriptives are needed, free-form typing is used and correct punctuation should be added. 11. Select the Drug button. 12. Enter the Substance to be reported, the Amount, Units, Percentage (Pct), Total Weight and Total Weight Units. 13. Enter any adulterants or dilutants in subsequent Substance fields. 14. When all information has been entered, select the Update Result button. 15. The User can then proofread the rough draft of the report, and add any other information in the Result field. 16. If more than one exhibit has been analyzed, repeat the above steps after advancing the Record number. NOTE: If multiple exhibits are found to contain the same controlled substance, it may be of value to the user for the report to include the total or aggregate weight of the multiple exhibits. This can be done by entering all data for the last exhibit number, entering all relevant exhibit numbers in the Aggregate Exhibits box, entering the aggregate weight and units, and then clicking the Update Result button.
17. Enter all Examinations that were performed for each exhibit. The defaulted examinations for Drug Cases are Color Test, Gas Chromatography, and Gas Chromatography/Mass Spectrometry. To add or delete examinations, click on the Examinations button, click the Clear Examinations button, click on each test performed from the list on the left, and then click the Accept button. 18. Enter all Billing information. The defaulted billing charges are one Exploratory Qualitative (302) and one Gas Chromatography/Mass Spectrometry (305). Additional charges are added by using the drop down pick list and entering the corresponding number of charges for that examination. Billing is entered on a per exhibit basis. 19.When all data has been entered, click the Exit button to return to the Results menu, or enter a new FL# to enter data for additional cases. B. 1. 2. 3. 4. Results Entry (Marihuana Cases)
The user completing the case must be logged into Drug LIMS. From the Results menu, select Results Entry. Bar code scan or manually type in the desired FL#. Enter the number, description of the evidence submitted, and weight and units of the evidence bag(s), box(es), etc. The drop down lists may be used for this operation. Refer to the reporting results section in Analysis and Reporting of Casework. 5. Enter the exhibit number. 6. The Analysts initials will appear in the Analyst box. The default is the User currently logged into Drug LIMS. 7. The Requesting Agency is defaulted from the evidence registration process, but should be verified before proceeding. 8. Enter the number of items tested. NOTE: Entering one will create the item container in singular form, any other entry will result in a plural format. 9. Enter exhibit description and container. The drop down lists may be used for this operation. A comma will be placed between the two descriptives, if only one adjective is needed, use only the first description box. If additional descriptives are needed, free form typing is used and correct punctuation should be added. 10. Select the Marihuana button. 11. The Substance to be reported, marihuana is the default, the Total Weight and Total Weight Units are entered. 12. When seed germination is requested, Click the Seeds box if seeds were present, and click the Germinate box if the seeds were capable of germination. If seeds were capable of germination, then the Net Weight and Net Units are entered. 13. When all information has been entered, select the Update Result button. 14. The User can then proofread the rough draft of the report, and add any other information in the Result field. 15. If more than one exhibit has been analyzed, repeat the above steps after advancing the Record number. NOTE: If multiple exhibits are found to contain marihuana, it may be of value to the user for the report to include the total or aggregate weight of the multiple exhibits. This is done by entering all data for the last exhibit number, entering all relevant exhibit numbers in the Aggregate Exhibits box, entering the
aggregate weight and units, and then clicking the Update Result button. 16. Enter all Examinations that were performed for each exhibit. The default examinations for Marihuana Cases are Color Test, Microscopic Test, Gas Chromatography/Mass Spectrometry, and Seed Germination (when seeds are present). To add or delete examinations, click on the Examinations button, click the Clear Examinations button, click on each test performed from the list on the left, and then click the Accept button. 17. Enter all Billing information. The default billing charges are one Marihuana/Cannabinoids (308), one Gas Chromatography/Mass Spectrometry (305), and one Marihuana Seed Germination (309), (when conducted). Additional charges are added by using the drop down pick list and entering the corresponding number of charges for that examination. Billing is entered on a per exhibit basis. 18. When all data has been entered, click the Exit button to return to the Results menu, or enter a new FL# to enter data for additional cases. C. 1. 2. 3. 4. Results Entry (Product Identification) The user completing the case must be logged into Drug LIMS. From the Results menu, select Results Entry. Bar code scan or manually type in the desired FL#. Enter the number, description of the evidence submitted, and weight and units of the evidence bag(s), box(es), etc. The drop down lists may be used for this operation. Refer to the reporting results section in Analysis and Reporting of Casework. Enter the exhibit number. The Analysts initials will appear in the Analyst box. The default is the User currently logged into Drug LIMS. The Requesting Agency is defaulted from the evidence registration process, but should be verified before proceeding. The total number of items (tablets/capsules) are entered into the Number of items field. NOTE: Entering one will create the item container/product form in singular form, any other entry will result in a plural format. Enter an item description and container. This entry is actually a description of the product form (capsule/tablet/etc.). The drop down lists may be used for this operation. Select the Product ID button. Do not make an entry in the Product Name field. Press the Product Search button. A new form will appear. If the product is known, select it from the Find Product drop down list. When the desired product is found, press the Send Selection to Report button. If the product is NOT on the list, a new entry can be created by typing the unique identifier (logo) into the Unique Identifier field. Copy (Ctrl-c) and paste (Ctrl-v) this unique identifier into the Logo and Find Product fields. Enter all known information into blank fields and press the Send Selection to Report button. Enter data into remaining fields (Total Weight and Units), and click the Analyzed box if a mass spectrum was obtained, and what substance(s) was identified. When all information has been entered, select the Update Result button.
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5. 6. 7. 8.
9.
10. 11. 12. 13.
14. 15.
16. The User can then proofread the rough draft of the report, and add or delete any other information in the Result field. 17. If more than one exhibit has been analyzed, repeat the above steps after advancing the Record number. 18. Enter all Examinations that were performed for each exhibit. The default examinations for Product ID Cases are Logo, Gas Chromatography/Mass Spectrometry (when MS performed). To add or delete examinations, click on the Examinations button, click the Clear Examinations button, click on each test performed from the list on the left, and then click the Accept button. 19. Enter all Billing information. The default billing charges are one Tentative Identification (312) and one Gas Chromatography/Mass Spectrometry (305), when performed. Additional charges are added by using the drop down pick list and entering the corresponding number of charges for that examination. Billing is entered on a per exhibit basis. 20. When all data has been entered, click the Exit button to return to the Results menu, or enter a new FL# to enter data for additional cases. D. Case and Billing Reports
This function is used for completing casework and printing the final report and the billing report. The user completing the case must be logged into the Drug LIMS. From the Results menu, select Case and Billing Reports. Bar code scan or manually type in the desired FL#. Enter the Completion Date (the year is defaulted). Enter the Reviewer (drop down list). Enter any other important case information in the Other References box. This information includes the arresting officer badge number(s), requesting department, resubmittal date, etc. 7. The report may be previewed by pressing the Preview Report button. NOTE: Do NOT print the report from this screen. 8. To print the final report, press the Print Report button. A hard copy of the report is printed and a copy of the report is sent to a default report file in Word format. 9. Print the Billing Report by pressing the Print Billing Report button. This report may also be previewed by pressing the Preview Billing Report button (NOTE: Do NOT print the billing report from this screen). When printing the billing report, the User will be prompted that the Drug LIMS is updating tables, and should the User proceed. Answer yes and update tables only when the billing report is correct. 10. If the case is shared with another chemist, the chemist finalizing the report should print the billing report when the final report is printed. 11. Repeat steps 3-9 for reports to be printed. 12. When finished, press the Exit button. E. Review Cases 1. 2. 3. 4. 5. 6.
This function may only be used by Authorized Reviewers such as a supervisor. Once a report is reviewed and countersigned, the reviewer will approve the case. A reviewed date is entered and a non-official copy of the report is sent to the Dallas County DAs computer server for their use. NOTE: Only DPD cases are sent to the DAs server. VI. Queries
This function is used to find cases by a number of different entries. Two views are allowed for this screen: Form View and Datasheet View. Form View is ideal for looking at a single record. Datasheet View is best suited for multiple records. A disadvantage of the latter is that to view all information the User must scroll the screen horizontally to see all fields. Scrolling through the records is accomplished by using the record selectors at the top or bottom of the screen. To find specific cases: 1. From the Main Switchboard, select Queries button. 2. Select the Find Case button. 3. Select the Filter By Form button. 4. Enter the desired information to be searched into any of the fifteen (15) fields. NOTE: More than one field may be entered. 5. Select the Apply Filter button. 6. When finished select the Exit button. NOTE: If an item to be searched might have more than one entry, for example, a defendant with the last name of Smith, then enter the last name and an asterisk in quotation marks ( Smith,* ) and select the Filter by Form button. VII. Utilities
This function is used to change the Users password and to clear all transfer tables. To change password: 1. Select the Change Password button 2. Enter Old Password 3. Enter New Password 4. Confirm New Password 5. Select OK Drug LIMS will then restart and the User will be asked to log back into the system. NOTE: The Clear Transfer Table should only be used by the Evidence Registrar or Supervisor, and only when there is not a transfer of cases being preformed. VIII. Log Out This function is used to exit out of the Drug LIMS.
1. Select the Log Out button (this will take the User to the Login Screen). 2. Select the Quit button to exit the system and Access.
COURT TESTIMONY, DEPOSITIONS, and other LEGAL PROCEEDINGS 1. General 1.1. The Dallas County Institute of Forensic Sciences is an independent laboratory which provides services on a fee for services basis under direction of the Dallas County Commissioners Court and the Director of the Institute. 1.2. As such, the Institute is not part of any police agency, the Dallas County Sheriffs Office, or the District Attorneys Office. 1.3. Most cases submitted to the Laboratory are forensic in nature and employees may reasonably be expected to provide testimony in legal proceedings regarding facts and opinions related to submitted evidence. 1.4. As outlined in the mission statement of the Institute, it is the goal of Institute staff to perform forensic laboratory analyses with accuracy, integrity, and reliability and to present written and oral results of analyses in a complete and impartial manner. 2. Fact Testimony 2.1. All employees of the Forensic Chemistry Section are subject to call for fact testimony by virtue of their employment with the Institute. 2.2. This type of testimony is used for building a record at trial concerning events occurring prior to trial such as receipt and release of evidence, custody of evidence, typing of reports, etc. 2.3. Employees called as fact witnesses must limit their testimony to personal actions taken (receipt and release of evidence, record generation, lab number assignment, typing, etc.). 2.4. Use of laboratory records by fact witnesses shall be limited to identification of applicable records and actions personally conducted by the employee. 2.5. The fact witness should not respond to questions about legal or scientific aspects of the case; these should be referred to other witnesses. 3. Introducing a Business Record 3.1. Laboratory staff may be called to introduce a laboratory report as a business record. 3.2. This typically occurs when the testifying staff did not perform work on the case under discussion. 3.3. The witness will answer a standard set of questions designed to establish the case report as a business record. 3.4. Then the witness will read the report into the record. 4. Expert Witness Testimony 4.1. Most testimony performed by analysts and supervisors will blend fact testimony with opinion or expert testimony. 4.2. Testifying staff are expected to maintain proficiency in the following areas: 4.2.1. A thorough understanding of applicable Texas and federal laws. 4.2.2. A broad general knowledge of natural science as applicable.
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4.2.3. An appropriate subject matter expertise in the employees discipline. 4.2.4. Knowledge of the legal requirements for maintaining chain of custody of evidence and for ensuring care, custody, and control of evidence and official records of their analyses. 4.2.5. Mastery over the scientific and technical aspects of the analytical methods used and the legal and scientific certainty of the validity of their results. 4.2.6. The ability to defend their analytical findings under court examination with clear and concise testimony using acquired forensic skills. 4.3. Until such time as an employee is considered acceptable by the Section Chief to give courtroom testimony, he shall not provide expert testimony in those cases in which he has prepared the report but shall refer such cases to the designated analyst or supervisor who will have co-signed the case report. 4.4. Expert witness testimony will be limited to the analysts area of expertise and will be further limited to testimony in cases in which the analyst actually performed work, work was performed under their supervision, or testimony is provided with approval of a supervisor. 4.5. The analyst is responsible for testimony concerning laboratory policy or interpretation of the law only as it pertains to the analytical conclusion reached in the specific case at trial. 4.6. Because expert knowledge is not subject to subpoena, analysts cannot be required to provide testimony in any case in which they did not perform analyses. 4.7. Permission to conduct outside employment requires prior permission of the Section Chief forwarded up to the Institute Director; the policy relating to outside employment is covered in the Dallas County Code. 4.8. An employee providing testimony as part of his County duties, must produce a subpoena unless testimony is provided for the Dallas County District Attorneys Office or meets other standing arrangement. 4.8.1. Pre-arrangement has been made with the Dallas County District Attorneys Office to honor telephone or fax requests in order to avoid the expense and inconvenience of issuing subpoenas for Institute employees. 4.9. Individuals leaving the building to go to Court should check out with the Section Secretary or Evidence Registrar. 4.10. All requests including subpoenas for testimony which is unrelated to the analysis of a specific Institute case must be reviewed prior to trial with a supervisor or Institute Director in the absence of a supervisor. 4.11. The individual receiving a subpoena is responsible for advising the requestor of applicable preparation, testimony, and travel fees to be charged by Dallas County related to the subpoena; written agreement to pay fees should be received from the requestor prior to the date of the proceeding. 4.11.1. Disagreement about payment of fees should be brought to the attention of a supervisor and the Civil Section, Dallas County District Attorneys Office, prior to the proceeding. 4.11.2. The Dallas County Commissioners Court has ordered that payment is required in advance (two hours minimum) for all services provided to private attorneys and any costs in excess of the minimum are payable at the time service is rendered.
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4.11.3. At the discretion of the Director, payment may be required before any service is rendered. 5. Testimony in the Capacity of Supervisor 5.1. The Eighth District of Texas, Court of Appeals in the case of Antoine Delano Caw v. the State of Texas (No: 08-92-00155-CR) has determined that chemists employed by the Dallas County Forensic Laboratory do not meet the definition of law enforcement personnel. Therefore, test results sponsored by a supervisor are admissible as a business records exception to the hearsay rules. 5.2. A supervisor usually enters the report as a business record. He then provides testimony regarding education and training of the analyst and their oversight of the analyst. After review of case file documentation, the supervisor can also testify as an expert witness based upon his own evaluation of data in the case file. 6. Dress 6.1. Professional business attire should be worn to all legal proceedings. 6.2. This usually includes a tie and/or jacket for men and dress, skirt, or pantsuit for women. 6.3. Shorts, jeans, and tee shirts are not appropriate. 6.4. Physical appearance should be neat and professional. 6.5. Dress code for federal may be more stringent; direction is usually provided with the subpoena if applicable. 7. Review of Testimony 7.1. Testimony of staff will be reviewed annually as described in the Quality Management Program. 8. General Guidelines for Testimony 8.1. Tell the truth at all times. 8.2. If you do not know or are unsure of an answer, clearly state this; you may state that you are not trained in this area, refer the question to a supervisor, etc. 8.3. Remain composed and professional; treat all parties with respect. 8.4. Keep on sound scientific ground; stay within your area of expertise. 8.5. Be prepared; review case prior to trial. 8.6. Understand the difference between possibility and probability. 8.7. Do not answer questions you do not understand. Ask for questions to be repeated or rephrased. 8.8. Do not provide answers to unasked questions. 8.9. Explain scientific concepts in clear laymans terms; avoid use of jargon. 8.10. Speak to the jury (jury trial) or judge (trial before the court). 8.11. When problems or confusion arise, speak to the Judge. 8.12. Speak slowly and distinctly. Speak in a tone that can be heard by the jury and court reporter.
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory 3 Testimony Version 2.0
8.13. Discuss any concerns with your supervisor.
Testimony Version 2.0
This is a non-controlled copy of a controlled document.
QUALITY MANAGEMENT I. General: A. Quality control and quality assurance are integral parts of the forensic activities performed at the Institute. B. Quality processes are described in detail in the Quality Manual and procedure specific quality issues are noted in procedures. C. Results of quality control samples may be found in the case file and/or in the Controlled Substances Laboratory. II. Critical Reagents: A. Internal Standard Solution 1. Preparation: Refer to Procedures Manual. 2. Frequency of Calibration: As made. 3. Tolerance: Refer to Internal Standard QC Log 4. New internal standard solution is compared to previous solutions. The peak area is monitored by gas chromatography. The % difference between the new I.S. with the old I.S. must be within 5.0 %. 5. Documentation of Acceptability: Refer to the Internal Standard QC Log 6. Corrective Action: If tolerances are not within expected range, the solution should be made again or an explanation must be made on the Comments column of the Internal Standard QC Log and reviewed with a supervisor. B. Stock Standards and Quality Control Solutions 1. Preparation: Refer to the Procedure Manual. 2. Frequency of Calibration: As made. 3. Tolerance: Refer to the Procedure Manual. 4. Documentation of Acceptability: a) Standards: Refer to Calibration Curve/Response Factor Log. b) QC Solutions: Refer to Procedure Manual. 5. Corrective Action: a) Standards: Refer to the Procedure Manual. b) QC Solutions: Refer to Procedure Manual. C. Color Test Reagents 1. Preparation: Refer to the Procedures Manual. 2. Frequency of Calibration: Refer to the Monthly Test Reagent Check List. 3. Tolerance: Refer to the Monthly Test Reagent Check List. 4. Documentation of Acceptability: Refer to the Monthly Test Reagent Check List. 5. Corrective Action: Refer to the Monthly Test Reagent Check List. III. Case Review: A. By signing a report, staff acknowledges that they have conducted an administrative case review.
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory 1 Quality Management Version 2.2
B. Primary technical review is conducted by the primary analyst. C. Technical review is also conducted by a supervisor or any other signer of the case report. D. Peer/technical review is also conducted by a non-signing analyst for 20% or six (whichever is least ) cases per analyst per month. E. During the early stages of training, a report may be signed by the training analyst, the instructor, and a supervisor. F. Components of case review are described in the Institute Quality Manual and in the Guidelines for Reviewing Drug Cases resource document. IV. Controlled Substances Quality Committee: A. The Controlled Substances Quality Committee is made up of two or more Chemists selected by the Controlled Substances Supervisor. Criteria for serving on the Committee includes but is not limited to a broad understanding of overall laboratory function, organization and communication skills, attention to detail, etc. B. Quality Committee responsibilities include 1. Review logs and records for completeness and consistency with policy prior to filing. 2. Maintain awareness of processes and procedures used in the Laboratory and identify departures from written procedures or good laboratory practice. 3. Recommend changes to procedures, policies, and processes to ensure and improve Laboratory quality. C. The Quality Committee functions independently within the Laboratory with reference to quality process review. 1. When quality issues are identified that cannot be addressed through routine policies and procedures, the Quality Committee member should immediately bring this to the attention of the Controlled Substances Supervisor, Lab Chief, and/or the Institute Quality Manager. a) Disagreement regarding appropriate action will be referred to the Executive Committee. D. All Controlled Substances staff is expected 1. To actively participate in quality processes as outlined in the Institute Quality Manual and Laboratory Quality Manual. 2. To run and evaluate appropriate QC as noted in analytical procedures. 3. To participate actively with the Quality Committee to resolve noted issues and to further quality processes in the Laboratory. V. Reliability of Standards, Controls, and Critical Reagents: A. Refer to the Controlled Substances Procedure Manual for specific information regarding stability of standards, controls, and reagents. B. In general, standards, controls, and critical reagents are considered acceptable for use as long as results meet assay acceptance criteria.
Quality Management Version 2.2
VI. Use of Positive and Negative Controls A. Positive quality control samples are run each day an assay is performed and evaluated as outlined in each procedure. 1. Results are summarized on the applicable GC or GC/MS Quality Control Log; the log and instrument output are filed in the applicable GC or GC/MS Logbook. B. A negative control is run each day a GC or GC/MS is used. 1. On a rotating basis, a procedural blank from one procedure is extracted and analyzed by GC. a) Results are summarized on the applicable GC Quality Control Log; the log and instrument output are filed in the applicable GC QC Logbook. b) On a weekly basis, rotate between sodium bicarbonate (NaHCO3) extract, sodium hydroxide (NaOH) extract, and ethanol (EtOH) with IS; prepare negative controls weekly. (1) For example: (a) Week 1 NaHCO3 extract, name NEGCTRLNaHCO3, use method METH.M (b) Week 2 EtOH with IS, name NEGCTRL-EtOH, use method HEROIN.M (c) Week 3 NaOH extract, name NEGCTRL-NaOH, use method METH.M 2. Solvents used for GC/MS are analyzed as a negative control for GC/MS. a) Results are summarized on the applicable GC/MS Quality Control Log; the log and instrument output are filed in the applicable GC/MS QC Logbook. (1) A fresh sample of solvent is analyzed. (a) Methylene chloride with internal standard, name NEGCTRL-MeCl2, use method NEGCTRL.M (b) Methanol, name NEGCTRL-MeOH, use method DRUGANA.M 3. An acceptable negative control contains no identifiable drug substance; however, it may contain a solvent front and minor non-drug peaks. The GC chromatogram should not contain peaks with retention times near that of routinely tested drugs. a) The mass spectrum for all non-solvent peaks on the GC/MS negative control total ionization chromatogram must be evaluated by the analyst to determine if a peak is drug related. (1) If a peak is not drug related it will be lined through and initialed by the analyst. (2) If a peak is drug related, troubleshooting will be conducted to determine the source of the drug, and casework will not be performed on that instrument until this issue is resolved.
b) If the GC chromatogram for the negative control contains non-solvent peaks, the negative control will be run by GC/MS to determine whether the peak is drug related. (1) If a peak is not drug related it will be lined through on the total ionization chromatogram, initialed by the analyst, and retained with the GC negative control. (a) A recurring minor peak in the negative control (based upon retention time) needs to be run by GC/MS only once. (2) If a peak is drug related, troubleshooting will be conducted to determine the source of the drug, and casework will not be performed on the applicable GC until this issue is resolved. c) Troubleshooting peaks in the Negative Control (1) The analyst will advise a supervisor that peaks are present in a negative control, whether peaks are drug or non-drug related, and outline the steps that will be taken to evaluate the situation. (a) Evaluation may include but is not limited to reinjecting the negative control, analyzing a new negative control, performing instrument maintenance and/or repair, etc. (2) The supervisor and the analyst will - in all cases - evaluate the sample(s) analyzed immediately prior to a negative control containing drug substance to determine whether these samples should be reevaluated and/or reanalyzed. VII. Housekeeping: A. Good housekeeping is the responsibility of Analysts, Evidence Registrars, and others working in the Controlled Substances Laboratory. B. Areas to be cleaned are designated as Workstations 1- 5 for room 206 and Workstations 1-4 for room 401 (refer to maps located on the Cleaning Checklist form). 1. These areas are sample prep areas used by the chemists. C. Additional areas that also require cleaning on a routine basis include: hoods, office and designated clean areas, and around instrumentation and computers. D. Each week cleaning will be documented on the Drug Laboratory Cleaning Checklist. E. Routine Cleaning (not recorded on Cleaning Checklist) 1. After each exhibit or case is completed, the chemist is responsible for wiping down his or her bench area with clean paper towels or kimwipes to remove any residue material from the previous exhibit or case. 2. If the bench paper is soiled, replace it. 3. Wipe off balances, spatulas, scalpels after each exhibit or case. F. Daily Cleaning (not recorded on Cleaning Checklist) 1. Put away glassware 2. Put away reagents 3. Discard extraction tubes and autosampler vials as needed
G. Weekly Cleaning (recorded on Cleaning Checklist) 1. Clean workstation thoroughly 2. Re-stock supply drawers 3. Check reagent levels 4. Dust or vacuum around instruments and computers 5. Clean hoods 6. Clean office or designated clean area 7. Check the biohazard boxes, they should not exceed 20 lbs. (~3/4 full) 8. Check chemical waste bottles, contact Deputy EHS manager for disposal H. 3. Monthly Cleaning (recorded on Cleaning Checklist) 1. Vacuum behind instruments VIII. Testing Validation: A. Validation and verification of new testing methods is summarized in the Quality Manual. B. The Supervisor and/or Section Chief will typically design a specific testing protocol to validate the accuracy of a new method or to verify the accuracy of a method transferred from one instrument to another. 1. Transfer of a Method to New Instrumentation a) Proper calibration and operation of new instrumentation are verified and documented using the applicable instrument calibration process. b) Analytical results are compared between the new and old instrumentation. (1) Where possible, side by side operation of old and new instrumentation is compared using the same samples or extracts. c) Standards, internal standards, and quality control samples should meet their respective criteria described in the analytical procedure. d) Test specimens should also be run and should meet acceptability criteria established for quality control samples or similar proficiency test results. e) Replicate analytical runs should be made on a single day and also compared between multiple days; criteria should meet those established for standards, internal standards, and quality controls; test specimens should meet acceptability criteria established for quality control samples or similar proficiency test results. f) As applicable, analysts are trained in operation of new instrumentation. 2. Development of a New Analytical Method a) A new analytical method may be proposed based upon literature, other established procedures, or knowledge and experience of the materials to be analyzed. b) The target sensitivity and linear range is established based upon expected sample conditions and instrument performance. c) Standard curves are run to assess the applicability of the proposed analytical technique in comparison to the targeted analytical needs.
d) A formal procedure is written. e) The method is evaluated for interference from expected matrices or cooccurring substances. f) The method is evaluated for stability by repeating analysis of single prepared samples and duplicate preparation of samples both within day and between day. g) Variability of standards, internal standards, quality control samples, and specimens are evaluated with respect to literature values and/or similar established procedures. h) Criteria for acceptable standard curves, internal standards, and/or quality control samples are established. i) The method is revised and revalidated as necessary. j) Applicable analysts are trained, and results are reviewed for stability within analyst. k) Method results may be evaluated against external standards or specimen results from reference laboratories; criteria for acceptability will be similar to that established for quality control samples.
ABBREVIATIONS LIST
The following is a list of abbreviations used in the Drug Laboratory section. The abbreviations can be used in any combination of upper and lower case with or without periods. Common chemical abbreviations are equally acceptable and are not listed here. Word or phrase 6-monoacetylmorphine alprazolam amount blank clear deionized water dilution do not report evidence exhibit forensic laboratory number gross weight Institute of Forensic Sciences internal standard large liquid lysergic acid diethylamide marihuana material methamphetamine miscellaneous net weight no controlled substance no reaction petroleum ether Physicican's Desk Reference powder precipitate quality control residue retention time sealed cardboard box small standard syringe unknown volume weight with without Abbreviation mam alpz amt blk clr DI H2O dil DNR evid ex FL# gwt ifs I.S. lg liq lsd mj mat meth misc net wt NCS NR pet ether PDR pwdr ppt qc res rt scb sm std syr unk vol wt w/, c (with a line over it) w/o
Abbreviations Verion 2.0
Drug and Procedure Cross Reference

Drug
acetaminophen alprazolam amphetamine barbituates boldenone caffeine carisoprodol chlordiazepoxide clonazepam cocaine codeine diazepam diethylpropion ethchlorvinyl ethosuximide fenfluramine flurazepam ghb heroin hydrocodone hydromorphone ibuprofen ketamine lorazepam lsd MDA MDEA MDMA meperidine meprobamate methamphetamine monoacetylmorphine morphine mushrooms/cactus buttons Dallas County Institute of Forensic Sciences Controlled Substances Manual
Procedure
acidic direct dilution alkaline acidic miscellaneous materials acidic acidic weak alkaline direct dilution weak alkaline alkaline direct dilution alkaline acidic acidic alkaline direct dilution miscellaneous materials direct dilution alkaline weak alkaline acidic weak alkaline direct dilution lsd alkaline alkaline alkaline alkaline acidic alkaline direct dilution weak alkaline/direct dilution miscellaneous materials Drug and Procedure Cross Reference Version 2.0
nandrolone olanzapine oxandrolone oxazepam oxycodone oxymetholone phencylidine phendimetrazine phentermine prazepam propoxyphene salicylates stanolone stanozolol temazepam testosterone thc theophylline tiazolam valproic acid
miscellaneous materials direct dilution miscellaneous materials direct dilution alkaline miscellaneous materials weak alkaline alkaline alkaline direct dilution alkaline acidic miscellaneous materials miscellaneous materials direct dilution miscellaneous materials marihuana acidic direct dilution acidic
Dallas County Institute of Forensic Sciences Controlled Substances Manual
Drug and Procedure Cross Reference Version 2.0
FORMATION OF ACETYLATED DERIVATIVES Analysis of N-Hydroxy MDA Principle of Assay: In some situations, chemicals with reactive hydrogens do not chromatograph well due to adsorption and/or thermal break down in the inlet of a gas chromatograph (GC). Occasionally, analytes do not separate well enough by GC for their analysis. Under these circumstances, chemical derivatization may solve these chromatographic issues and allow successful analysis of the substances in question. Acetylation is one method of derivatization. Using acetylation, chemicals which contain active hydrogens (for example OH, -SH, and NH) are converted into esters, thioesters, and amides respectively. When acetic anhydride [(CH3CO)2O]is used as the acetylating agent, the reactive hydrogen is replaced with an acetyl group (CH3CO-). At the high temperatures encountered in GC and GC/MS, N-hydroxy-3,4methylenedioxyamphetamine (N-hydroxy MDA) undergoes pyrolytic disproportionation. One molecule of N-hydroxy MDA is oxidized to 3,4-methylenedioxyphenyl-2-propanone-2-oxime (oxime) with simultaneous reduction of another molecule of N-hydroxy MDA to 3,4 methylenedioxyamphetamine (MDA). As a result at least two chromatographic peaks are produced. Acetylation of N-hydroxy MDA results primarily in a diacetylated molecule which is stable during GC and GC/MS analysis. Whenever N-hydroxy MDA and the oxime are detected by standard analytical procedures, acetylation should be performed to accurately identify the substances present. Equipment: Gas chromatograph-mass spectrometer a methylsiloxane capillary column such as an HP-5MS, 30 m x 0.25 mm x 0.25 um film thickness and gas chromatograph-flame ionization detector with split/splitless injector containing a methylsiloxane capillary column such as an HP-1, 15m x 0.32 mm x 1.0 um film thickness (see GC/MS and GC Methods Notebooks in Drug Laboratory). Screw top culture tubes, 25 mL, 20 mm x 125 mm, screw cap with Teflon liner Screw top culture tubes, 15 mL, 16 mm x 125 mm, screw cap with Teflon liner Volumetric flasks, 10 mL, 25 mL, 500 mL Autosampler vials, 32 mm x 11 mm, Teflon lined seals and glass inserts Vortex mixer Centrifuge Test tubes Analytical balance Pipettes, various pH paper Reagents: Acetic anhydride, reagent grade or better
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory Acetylated Derivatives Version 2.0
n-Butyl chloride, reagent grade or better Naphthalene, reagent grade or better Sodium bicarbonate, reagent grade or better Methanol, reagent grade or better HCl, reagent grade or better Methanolic HCl (5%) Transfer 5 mL concentrated HCl and 95 mL of methanol to a 100 mL Erlenmeyer flask. Analytical standards Safety Precautions: The most common type of chemical or biological exposure in this type of laboratory is a splash to the skin or eye. Skin, mucous membranes, or eyes which have been splashed with commonly used chemicals or biologicals should be thoroughly washed for at least 15 minutes in cool tap water or eye wash station. Refer to appropriate MSDSs for additional chemical information. Report the incident immediately to a supervisor. Seek medical attention as necessary. Acetic anhydride is corrosive and flammable. Use in a hood. Do not breathe vapors. Avoid contact with skin, eyes, or mucous membranes. Acetic anhydride is sensitive to moisture and will decompose. n-Butyl chloride is flammable. Avoid contact with skin, eyes, and mucous membranes. Methanol is a flammable solvent. Avoid contact with skin, eyes, and mucous membranes. Hydrochloric Acid is corrosive. Avoid contact with skin, eyes, and mucous membranes. Internal Standard (I.S.) Solution Preparation: Internal Standard 0.5 mg/mL naphthalene Amount of Chemical 250 mg Final Volume 500 mL Solvent n-butyl chloride
Transfer 250 mg naphthalene to a 500 mL volumetric flask and bring to volume with solvent. Mix by inverting the flask. (Note: Naphthalene is used as internal standard instead of phenanthrene because incomplete acetylation of N-hydroxy MDA produces a chromatographic peak which interferes with phenanthrene). Calibration Standards: All solutions of stocks and standards are kept in the refrigerator. Allow the solutions to come to room temperature before using them. The concentration, amount, and volume of the calibration stock and calibration standards may vary proportionally depending on the availability of drug, and the expected concentration range of the drug. The following is provided as a guide:
Acetylated Derivatives Version 2.0
Acetylation Procedure Calibration Stock Amount of Drug (as base) 5.0 mg/mL Drug 50.0 mg
Step 1 Step 2 Acetylation Final Volume Add 1-2 mL acetic anhydride 10 mL DI water
Transfer 50.0 mg drug calculated as the free base to a 15 mL screw cap culture tube, add 1-2 mL acetic anhydride, cap, let stand 24 hours at room temperature. Evaporate to dryness. Transfer 10 mL of deionized water by volumetric pipette to the culture tube. Vortex and sonicate as needed. Calibration Procedure: 1. Choose a minimum of three appropriate calibration standards for the calibration curve based on the linear range required for the sample or drug to be analyzed. 2. Using a Hamilton syringe or volumetric pipette, place the appropriate amount of calibration stock into a 15 mL labeled, screw cap culture tube. Add deionized (DI) water to make a total volume of 5.0 mL. Add approximately 200 mg sodium bicarbonate and 5.0 mL internal standard solution. Calibration Standards 0.10 mg/mL 1.00 mg/mL 5.00 mg/mL 3. 4. 5. 6. Amount of Calibration Stock 0.1 mL 1.0 mL 5.0 mL Amount of DI H2O 4.90 mL 4.00 mL 0 mL Volume I.S. solution 5 mL 5 mL 5 mL
Cap the tubes and place on a rotator for 5 - 10 minutes. Centrifuge for 5 - 10 minutes on medium setting. Transfer an aliquot of the organic layer to an autosampler vial. Inject the extracted calibration standards into the GC using the appropriate instrument method. 7. Determine if the data are acceptable using the instructions found in the Calibration Curve/Response Factor Log. An acceptable calibration curve has a correlation coefficient, r2 > 0.995. If the curve is acceptable, file it in the Calibration Curve/Response Factor Log located in the Drug Laboratory. If not, seek supervisory assistance as needed.
Quality Control Standard: Ideally the quality control standard will be made from a different lot number or manufacturer than the calibration stock. At a minimum, it should be made from a different stock solution. A quality control standard must be successfully run on each day this procedure is used. The concentration, amount, and volume may vary proportionally depending on the availability of drug. The following is provided as a guide:
Acetylation Procedure QC Stock 5.0 mg/mL Drug Amount of Drug (as base) 50.0 mg Step 1 Step 2 Acetylation Final Volume Add 1-2 mL acetic anhydride 10 mL DI water
Transfer 50.0 mg drug calculated as the free base to a 15 mL screw cap culture tube, add 1-2 mL acetic anhydride, cap, let stand 24 hours at room temperature. Evaporate to dryness. Transfer 10 mL of deionized water by volumetric pipette to the culture tube. Vortex and sonicate as needed. Quality Control Procedure: 1. Using a volumetric pipette, place 1.0 mL of quality control stock and 4.0 mL deionized water into a 15 mL labeled, screw cap culture tube. Add approximately 200 mg sodium bicarbonate and 5.0 mL internal standard solution. QC Standard 1.0 mg/mL Drug 2. 3. 4. 5. Amount of QC Stock 1.0 mL Amount of DI water 4.0 mL Volume of I.S. solution 5.0 mL
Cap the tube and place on rotator for 5 - 10 minutes. Centrifuge for 5 - 10 minutes on medium setting. Transfer an aliquot of the organic layer to an autosampler vial. Inject the extracted quality control sample into the GC using the appropriate instrument method.
Evaluation Criteria: 1. If the QC sample is within the set limits of +/- 5% of the target concentration, proceed with analysis. 2. If the QC sample is not within the set limits of +/- 5% of the target concentration, additional action must be taken to obtain an acceptable QC result such as re-running QC, re-extracting and re-running QC, making new stock with re-extraction and reanalysis, recalibrating (run curve), and/or consulting a supervisor. Color Testing: See the section on Color Testing. Spot tests will usually be run on each item of nonpharmaceutical evidence within one case up to the applicable statutory weight limit. Color testing may be performed on pharmaceutical evidence as applicable. Analytical Procedure: 1) Powdered Samples, Hard Chunky Powders, and Other Solids: a) Obtain the total weight of the exhibit to be analyzed and record on the Drug Protocol Worksheet. b) Create a representative specimen by taking a sample from many or all of the solid pieces
in the exhibit. If powder is also present, remove a sample and add to the material scraped from the solid pieces. Pulverize the resulting composite sample and analyze as described. c) GC/MS Identification i) Transfer approximately 10 mg of material to 1 mL methanol in a test tube. (1) The analyst will vary the amount of material and methanol as needed to obtain an acceptable mass spectrum. ii) Vortex and centrifuge as necessary. iii) Transfer an aliquot to an autosampler vial and inject on the GC/MS using BLANK.M method. A chromatogram displaying MDA and the oxime indicates the presence of N-OH MDA. iv) N-hydroxy MDA decomposes on the injection block to MDA and the oxime, therefore acetylation is required to provide definitive identification by GC and GC/MS. v) Transfer 50.0 mg sample to a 15 mL screw cap culture tube, add 1-2 mL acetic anhydride, cap, let stand 24 hours at room temperature. Evaporate to dryness. vi) Reconstitute with methanol. vii) Inject an aliquot on the GC/MS using BLANK.M method. d) GC Quantitation i) Weigh approximately 10 - 50 mg of the material to be analyzed, and record the weight on the Drug Protocol Worksheet. ii) Transfer sample to a 15 ml labeled, screw cap culture tube. iii) N-hydroxy MDA decomposes on the injection block to MDA and the oxime, therefore acetylation is required. iv) Transfer 50.0 mg sample to a 15 mL screw cap culture tube, add 1-2 mL acetic anhydride, cap, let stand 24 hours at room temperature. Evaporate to dryness. v) Add 5.0 mL of deionized water and approximately 200 mg sodium bicarbonate to the culture tube. vi) Add 5.0 mL of internal standard solution. vii) Cap the tube and place on rotator for 5 - 10 minutes. viii) Centrifuge for 5 - 10 minutes on medium setting. ix) Transfer an aliquot of the organic layer to an autosampler vial. x) Inject extracted sample into GC using the appropriate instrument method and calculate the results. Instrument Parameters: See GC and GC/MS Methods Notebooks located in the Drug Laboratory. Reporting Criteria: 1. Report both the quantity and the identity of the drug when the concentration of the drug is within +/- 20% of the calibration curve on the GC and the GC/MS is complete. It may be necessary to dilute extracted samples to bring them within the curve range. 2. Report the amount of the drug as insufficient for quantitation when the drug concentration is below 20% of the calibration curve on the GC and the GC/MS is complete.
Calculations: 1. Dilutions a. In any situation in which a dilution is made, multiply the concentration of drug (mg/mL) by the dilution factor. A notation relating to how dilutions were made must be made in the case file. 2. Materials Analyzed by Weight: a. concentration of drug (mg/mL) x volume of internal standard solution used for extraction (mL) x total exhibit weight (mg)/extracted sample weight (mg) = mg of drug in the exhibit as the free base 3. Percent drug in the exhibit: a. mg of drug in the exhibit/total weight of the exhibit x 100 = % of drug in the exhibit
Training Notes:Acetylation Procedure N-hydroxy MDA often appears as round unmarked white tablets or round green speckled tablets. Weight of tablets is often about 500 mg. 2. Whenever MDA and the oxime are identified in standard GC or GC/MS runs, this procedure should be performed to determine whether N-hydroxy MDA is present and the source of the MDA and oxime. 3. Acetic anhydride is moisture sensitive. It is also volatile and reactive; it should be used in the hood. 4. Since N-hydroxy MDA has two reactive hydrogens (the amine and hydroxyl), it will form a diacetyl derivative. Incomplete derivatization will produce both the monoacetyl and diacetyl products which will appear as two chromatographic peaks. 5. Liquids should be evaluated for suitability. Water, alcohols, and other liquid vehicles containing reactive hydrogens will compete with the sample for acetylation usually resulting in poor acetylation of the sample. Liquids may be taken to residue and then acetylated; however, care should be taken to remove water or other solvent residue prior to acetylation. 1.
ACIDIC DRUG ANALYSIS Principle of Assay: This method describes the analysis of drugs which are acidic in nature; often these substances contain an hydroxyl (-OH) or carboxylic acid (-COOH) functional group. Some neutral drugs may also be analyzed using this procedure. Drugs are identified using an analytical technique which provides conclusive identification of the drug. These techniques include gas chromatography/mass spectrometry (GC/MS) and infrared spectroscopy (IR). The acidic drug is dissolved in water and the pH of the solution is made acidic (approximately pH 3) using 25% HCl. In the acidic aqueous solution, the acidic drug is converted from the ionic salt form to the nonionic free acid form. The free acid form of the drug is quantitatively extracted into an organic solvent such as chloroform containing phenanthrene as the internal standard. Quantitative analysis is performed using gas chromatography (GC). Specific details of quantitation for each acid drug and its linear range of quantitation may be found in the Calibration Curve/Response Factor Log for each GC. Examples of acidic drugs include the barbiturates, meprobamate, valproic acid, ethosuximide, salicylate, acetaminophen, ibuprofen, caffeine, carisoprodol, theophylline, ethchlorvynol, etc. Sample Requirements: Powders, solids, liquids, tablets, capsules, residues, and other types of drug evidence are suitable for analysis. Equipment: Gas chromatograph-mass spectrometer containing a methylsiloxane capillary column such as an HP-5MS, 30 m x 0.25 mm x 0.25 um film thickness and/or gas chromatograph-flame ionization detector with split/splitless injector containing a methylsiloxane capillary column such as an HP1, 15 m x 0.32 mm x 1.0 um film thickness (see GC/MS and/or GC Methods Notebooks in Drug Laboratory). Screw top culture tubes, 25 mL, 20 mm x 125 mm, screw cap with Teflon liner Screw top culture tubes, 15 mL, 16 mm x 125 mm, screw cap with Teflon liner Volumetric flasks, 10 mL, 25 mL, 2 liter Autosampler vials, 32 mm x 11 mm, Teflon lined seals and glass inserts Vortex mixer Centrifuge Test tubes
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory Acidic Drug Analysis Version 2.0
Erlenmeyer flasks, various Graduated cylinders, various Analytical balance Pipettes, various pH paper Reagents: Phenanthrene, reagent grade or better Chloroform, reagent grade or better HCl, reagent grade or better Methanol, reagent grade or better HCl, reagent grade or better Analytical standards Reagent Volume of Chemical Volume of deionized H2O 25% HCl 10 mL 30 mL In the hood, place 30 mL deionized water into a 50 mL graduated cylinder and carefully add 10 mL of concentrated HCl using a graduated cylinder. Transfer to a 50 mL Erlenmeyer flask for storage. Methanolic HCl (5%) Transfer 95 mL of methanol to a 100 mL Erlenmeyer flask, then add 5 mL concentrated HCl. Safety Precautions: The most common type of chemical or biological exposure in this type of laboratory is a splash to the skin or eye. Skin, mucous membranes, or eyes which have been splashed with commonly used chemicals or biologicals should be thoroughly washed for at least 15 minutes in cool tap water or eye wash station. Refer to appropriate MSDSs for additional chemical information. Report the incident immediately to a supervisor. Seek medical attention as necessary. Chloroform is a chlorinated solvent and is considered a human carcinogen. Use in a hood or in a well ventilated area. Avoid contact with skin, eyes, or mucous membranes. Hydrochloric acid is corrosive. Avoid contact with skin, eyes, and mucous membranes. Methanol is a flammable solvent. Avoid contact with skin, eyes, and mucous membranes.
Acidic Drug Analysis Version 2.0
Internal Standard Solution Preparation: Internal Standard Amount of Chemical Final Volume Solvent 0.5mg/mL phenanthrene 0.25 grams phenanthrene 500 mL chloroform Transfer 0.25 g phenanthrene to a 500 mL volumetric flask, and bring to volume with solvent. Mix by inverting flask. Calibration Standards: All solutions of stocks and standards are kept in the refrigerator. Allow the standards to come to room temperature before using. The concentration, amount, and volume of the calibration stock and calibration standards may vary proportionally depending on the availability of drug, the expected concentration range of the drug, and the needed volume of standard. The following is provided as a guide: Calibration Stock Amount of Drug (free acid) Final Volume Solvent
5.0 mg/mL Drug 50.0 mg 10 mL deionized water Transfer 50.0 mg drug calculated as the free base to a 10 mL volumetric flask, and bring to volume with deionized (DI) water. Mix by inverting flask. Sonicate as needed. Transfer to a 15 mL screw cap culture tube for storage. Calibration Procedure: 1. Choose a minimum of three appropriate calibration standards for the calibration curve based on the linear range required for the sample or drug to be analyzed. 2. Using a Hamilton syringe, and/or a volumetric pipette, place the appropriate amount of calibration stock into a 15 mL screw cap culture tube. Add deionized water to make a total volume of 5.0 mL. Calibration Standards 0.10 mg/mL 1.00 mg/mL 5.00 mg/mL 3. 4. 5. 6. Amount of Calibration Stock 0.10 mL 1.0 mL 5.0 mL Amount of DI H20 4.90 mL 4.0 mL 0 Volume I.S. solution 5 mL 5 mL 5 mL
Add 5 drops of 25% HCl and 5.0 mL internal standard solution. Cap the tubes and place on rotator for 5 - 10 minutes. Centrifuge for 5 - 10 minutes on medium setting. Transfer an aliquot of the organic layer to an autosampler vial. Note: n-butyl chloride will be on the top; chloroform on the bottom. 7. Inject the extracted calibration standards into the GC using the appropriate method.
8. Determine if the data are acceptable using the instructions found in the Calibration Curve/Response Factor Log. a. An acceptable calibration curve has a correlation coefficient, r2 > 0.995. b. If the curve is acceptable, file it in the Calibration Curve/Response Factor Log located in the Drug Laboratory. If not, seek supervisory assistance as needed. Quality Control Standard: Ideally, the quality control standard should be made from a different lot number or manufacturer than the calibration stock standard. At a minimum, it should be made from a different stock solution. A quality control standard must be successfully run on each day this procedure is used. The concentration, amount, and volume may vary proportionally depending on the availability of drug. The following is provided as a guide: QC Stock Amount of Drug (free acid) Final Volume Solvent 2.0 mg/mL Drug 20.0 mg 10.0 mL deionized water Transfer 20.0 mg drug calculated as the free base to a 10 mL volumetric flask, and bring to volume with deionized (DI) water. Mix by inverting flask. Sonicate as needed. Transfer to a 15 mL screw cap culture tube for storage. Quality Control Procedure: 1. Using a volumetric pipette, place 2.5 mL of quality control stock and 2.5mL deionized water into a 15 mL screw cap culture tube. Add 4 drops of 25% HCl and 5.0 mL internal standard solution. Amount of QC Stock 2.5 mL Amount of DI H20 2.5 mL Volume of I.S. solution 5.0 mL
QC Standard 1.0 mg/mL Drug
2. Cap the tube and place on rotator for 5 - 10 minutes. 3. Centrifuge for 5 - 10 minutes on medium setting. 4. Transfer an aliquot of the organic layer to an autosampler vial. 5. Inject the extracted quality control sample into the GC using appropriate instrument method. Evaluation Criteria: 1. If the QC sample is within the set limits of +/- 5% of the target concentration, proceed to with analysis.
2. If the QC sample is not within the set limits of +/- 5% of the target concentration, additional action must be taken to obtain an acceptable QC result such as re-running QC, re-extracting and re-running QC, making new stock with re-extraction and re-analysis, re-calibrating (run curve), and/or consulting a supervisor. Color Testing: See the section on Color Tests. Spot tests will usually be run on each item of nonpharmaceutical evidence within one exhibit up to the applicable statutory weight limit. Analytical Procedure: 1) Powdered Samples and Other Solids: a) Obtain the total weight of the material in the exhibit to be analyzed and record on the Drug Protocol Worksheet. b) Create a representative specimen by taking a sample from many or all of the solid pieces in the exhibit. If powder is also present, remove a sample and add to the material scraped from the solid pieces. Pulverize the resulting composite sample as necessary and analyze as described. c) GC/MS Identification i) Transfer approximately 10 mg of material to approximately 1 mL methanol in a test tube. (1) The analyst will vary the amount of material and methanol as needed to obtain an acceptable mass spectrum. ii) Vortex and centrifuge as necessary. iii) Transfer an aliquot to an autosampler vial and inject on the GC/MS using BLANK.M method. d) GC Quantitation i) Weigh approximately 10 - 50 mg of the material to be analyzed, and record the weight on the Drug Protocol Worksheet. ii) Transfer sample to a 15 mL labeled screw cap culture tube. iii) Add 1.0 - 5.0 mL of deionized water using a volumetric pipette and 1 - 5 drops of 25% HCl using a Pasteur pipette to the culture tube. iv) Add 1.0 - 5.0 mL internal standard solution. v) Cap the tube and place on rotator for 5 - 10 minutes. vi) Centrifuge for 5-10 minutes on medium setting. vii) Transfer an aliquot of the organic layer to an autosampler vial. viii) Inject extracted sample into GC using appropriate instrument method and calculate the results. 2) Tablets, Capsules, and Other Solid Pharmaceutical Preparations: a) Obtain the total weight of the material in the exhibit to be analyzed and record on the Drug Protocol Worksheet.
b) Create a representative sample for analysis and grind to a powder if necessary. i) Capsules typically a composite sample is used for analysis. ii) Tablets typically one tablet is selected for analysis. c) Logo Identification i) Refer to Pharmaceutical Analysis Procedure. d) GC/MS Identification i) Transfer an amount of sample that is equivalent to approximately 1 mg of drug to approximately 1 mL methanol in a test tube. (1) The analyst will vary the amount of material and methanol as needed to obtain an acceptable mass spectrum. ii) Vortex and centrifuge as necessary. iii) Transfer an aliquot to an autosampler vial and inject on the GC/MS using BLANK.M method. e) GC Quantitation i) Weigh approximately 10 - 50 mg of the material to be analyzed, and record the weight on the Drug Protocol Worksheet. ii) Transfer sample to a 15 mL labeled screw cap culture tube. iii) Add 1.0 - 5.0 mL of deionized water using a volumetric pipette and 1 - 5 drops of 25% HCl using a Pasteur pipette to the culture tube. iv) Add 1.0 - 5.0 mL internal standard solution. v) Cap the tube and place on rotator for 5 - 10 minutes. vi) Centrifuge for 5-10 minutes on medium setting. vii) Transfer an aliquot of the organic layer to an autosampler vial. viii) Inject extracted sample into GC using appropriate instrument method and calculate the results. 3) Liquid Samples: a) Liquid samples may be analyzed by weight or by volume. If the sample is to be analyzed by volume, a known volume of the liquid must be weighed to determine density. i) If the volume of liquid is small, it should be treated as a residue; see below. ii) Obtain the total weight of liquid in the exhibit to be analyzed and record on the Drug Protocol Worksheet. (1) If the liquid is to be analyzed by volume, record the total volume on the Drug Protocol Worksheet. iii) Mix the liquid well before sampling. iv) GC/MS Identification (1) Place approximately 1 part sample to 4 parts methanol in a 15 mL screw cap culture tube. (a) The analyst will vary the amount of material and methanol as needed to obtain an acceptable mass spectrum. (2) Transfer an aliquot to an autosampler vial. Analyze by GC/MS using BLANK.M method. v) GC Quantitation
(1) Transfer a known volume or weight of the sample to a 15 mL labeled screw cap culture tube. (2) If the liquid is aqueous, proceed to step 3. (a) If the composition of the liquid is unknown or organic, add three drops of methanolic HCl and evaporate the sample to dryness or to constant volume. (3) Based on the estimated drug concentration of the material, determine the extraction volume. The volumes of water and extraction solvent should be equal: (a) Add 1.0 5.0 mL of deionized water, 1-5 drops of 25% HCl, and 1.0 - 5.0 mL internal standard solution. (b) Cap the tube and place on rotator for 5 - 10 minutes. (c) Centrifuge for 5 -10 minutes on medium setting. (d) Transfer an aliquot of the organic layer to an autosampler vial. (e) Inject the extracted sample into GC using appropriate instrument method and calculate the results. 4) Residue Samples: a) Carefully rinse the residue into a container using methanol. b) GC/MS Identification i) Run a methanol blank before each residue sample using DRUGANA.M or BLANK.M methods and place in case file. ii) Place an aliquot of the methanol washing into an autosampler vial. Analyze by GC/MS using BLANK.M method. c) GC Quantitation i) Transfer the sample/methanol mixture to a 15 mL glass screw cap culture tube, add 13 drops of methanolic HCl, and evaporate to dryness. ii) Based on the estimated drug concentration of the material, determine the extraction volume. Add the same amount of water and extraction solvent: (1) Add 1.0 5.0 mL of deionized water to residue, 1-5 drops of 25% NaOH, and 1.0- 5.0 mL internal standard solution. iii) Cap the tube and place on a rotator for 5 - 10 minutes. iv) Centrifuge for 5 - 10 minutes on medium setting. v) Run a methanol blank before each residue sample using the same method used for the sample and place in case file. vi) Transfer an aliquot of the organic layer to an autosampler vial. vii) Inject the extracted sample into GC using the appropriate instrument method and calculate the results. (1) If it is necessary to dilute the residue, return to the original washing and dilute. viii) When analysis is complete return any extracted sample remaining in the autosampler vial to the original culture tube used for the extraction. (1) Remove and discard the aqueous layer from the culture tube. (2) Add 1-3 drops of methanolic HCl to the extracted sample to convert the drug to its more stable hydrochloride salt form. (3) Take the extracted sample to residue, in a laboratory fume hood. Close with
a screw cap. (4) Label the tube added by laboratory, include FL# and initials, and place this tube in the evidence bag along with the evidence. Instrument Parameters: See the GC and GC/MS Methods Notebooks located in the Drug Laboratory. Reporting Criteria: 1. Report both the quantity and the identity of the drug when the concentration of the drug is within +/- 20% of the calibration curve on the GC and the GC/MS is complete. It may be necessary to dilute extracted samples to bring them within the curve range. 2. Report the amount of the drug as insufficient for quantitation when the drug concentration is below 20% of the calibration curve on the GC and the GC/MS is complete. Calculations: 1. Dilutions a. In any situation in which a dilution is made, multiply the concentration of drug (mg/mL) by the dilution factor. A notation relating to how dilutions were made must be made in the case file. 2. Materials Analyzed by Volume: a. concentration of drug (mg/mL) x volume of internal standard solution used for extraction (mL) x total exhibit volume/extracted sample volume = mg of drug in the exhibit as the free base 3. Materials Analyzed by Weight: a. concentration of drug (mg/mL) x volume of internal standard solution used for extraction (mL) x total exhibit weight (mg)/extracted sample weight (mg) = mg of drug in the exhibit as the free base 4. Percent drug in the exhibit: a. mg of drug in the exhibit/total weight of the exhibit x 100 = % of drug in the exhibit
Training Notes: Acidic Procedure 1. If GC/MS results are negative when run on BLANK.M, run on alternative GC/MS methods such as LOWDOSE.M, LSD.M, etc. until satisfied that no controlled substance is present. 2. The free acid form of a barbiturate may not be soluble in water; therefore, these standards may be made in ethanol. For quantitation, the proper amount if ethanolic calibration standard is evaporated to dryness before proceeding to extraction. 3. Clarkes Analysis of Drugs and Poisons is a good source for determining the solubility of a particular drug standard. 4. Some drug combinations such as Fiorinal with codeine contain acid and alkaline drugs. One approach for this combination preparation is to first perform an alkaline extraction for identification of codeine and caffeine and quantitation of codeine. Then perform an acid extraction on the aqueous phase remaining after the alkaline extraction. To do this, add 25% HCl to the aqueous phase until the pH<3. Continue with the acid extraction by adding 5.0 mL internal standard solution. Qualitatively identify salicylate and butalbital in the organic phase by GC/MS. 5. Some pharmaceutical preparations of acid drugs, such as phenobarbital, are in time release form. To improve recovery it this case, allow the sample to sit overnight in 5.0 mL water and 5 drops of 25% HCL. In the morning, continue the assay by adding 5.0 mL internal standard solution and following the acid drug procedure. 6. A sonicator may be used to bring materials into solution. Care should be used regarding the length of time materials are left in the sonicator since heat may decompose some drugs. Typically, a sonication time of 5 minutes or less is used. 7. Valproic acid and ethchlorvynol are volatile. Solutions containing valproic acid or ethchlorvynol should be evaporated carefully and removed from the evaporation station as the tube reaches dryness. Due to their volatility, valproic acid and ethchlorvynol should be run using the volatiles GC/MS method.
ALKALINE DRUG ANALYSIS Principle of Assay: This method describes the analysis of drugs which are alkaline (basic) in nature; these substances often contain an amine (-NR2) functional group. Some neutral drugs may also be analyzed using this procedure. Drugs are identified using analytical techniques which provide conclusive identification of the drug. These techniques include gas chromatography/mass spectrometry (GC/MS) and infrared spectroscopy (IR). The basic drug is dissolved in water and the pH of the solution is made basic (approximately pH 10) using sodium hydroxide. In the basic aqueous solution, the alkaline drug is converted from the ionic salt form to the nonionic free base form. The free base form of the drug is quantitatively extracted into n-butyl chloride (for most drugs) or chloroform (for hydrocodone) containing internal standard, usually phenanthrene. Quantitative analysis is performed on the extracted drug using gas chromatography (GC). Specific details of quantitation for each alkaline drug and its linear range of quantitation may be found in the Calibration Curve/Response Factor log for each GC. Examples of drugs analyzed using the alkaline procedure include: Drug Internal Standard Extraction Solvent amphetamine phenanthrene n-butyl chloride codeine phenanthrene n-butyl chloride diethylpropion phenanthrene n-butyl chloride fenfluramine phenanthrene n-butyl chloride hydrocodone phenanthrene chloroform (dihydrocodeinone) MDA phenanthrene n-butyl chloride MDMA phenanthrene n-butyl chloride MDEA phenanthrene n-butyl chloride methamphetamine phenanthrene n-butyl chloride oxycodone phenanthrene chloroform pethidine (meperidine) naphthalene chloroform phendimetrazine phenanthrene n-butyl chloride phentermine phenanthrene n-butyl chloride Marked pharmaceuticals will usually be identified by logo match and GC/MS without quantitation. However when the penalty group or schedule of a marked pharmaceutical preparation is determined by dosage or preparation, sufficient testing should be performed for placement of the substance in a proper penalty group or schedule. Sample Requirements: Powders, solids, liquids, tablets, capsules, residues, and other types of drug evidence are suitable for
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory 1 Alkaline Drug Analysis Version 2.0
analysis using this procedure. Equipment: Gas chromatograph-mass spectrometer containing a methylsiloxane capillary column such as an HP5MS, 30 m x 0.25 mm x 0.25 um film thickness and/or gas chromatograph-flame ionization detector with split/splitless injector containing a methylsiloxane capillary column such as an HP-1, 15 m x 0.32 mm x 1.0 um film thickness (see GC/MS and/or GC Methods Notebooks in Drug Laboratory). Screw top culture tubes, 25 mL, 20 mm x 125 mm, screw cap with Teflon liner Screw top culture tubes, 15 mL, 16 mm x 125 mm, screw cap with Teflon liner Volumetric flasks, 10 mL, 25 mL, 2 liter Autosampler vials, 32 mm x 11 mm, Teflon lined seals and glass inserts Vortex mixer Rotator Centrifuge Sonicator Graduated cylinders, various Test tubes Analytical balance Hamilton syringes, 100 uL, 250 uL, 500 uL Pipettes, various pH paper Adjustable 10 mL Dispensette Reagent Bottle Erlenmeyer flask, 50 mL, 100 mL Mortar and Pestle Reagents: n-Butyl chloride, reagent grade or better Chloroform, reagent grade or better Naphthalene, reagent grade or better Phenanthrene, reagent grade or better Methanol, reagent grade or better HCl, reagent grade or better Sodium hydroxide, reagent grade or better Analytical standards Reagent Amount of Chemical Volume of deionized H2O 40% NaOH (w/v) 40 g NaOH 100 mL Transfer to a 250 mL Erlenmeyer flask. In a hood, immerse the flask in a cold tap water bath. USE CAUTION, this mixture will get extremely hot. Cautiously swirl the flask to dissolve. Cool and mix well. Reagent Volume of Chemical Volume of deionized H2O
Alkaline Drug Analysis Version 2.0
25% HCl 10 mL 30 mL In the hood, place 30 mL deionized water into a 50 mL graduated cylinder and carefully add 10 mL of concentrated HCl using a graduated cylinder. Transfer to a 50 mL Erlenmeyer flask for storage. Methanolic HCl (5%) Transfer 95 mL of methanol to a 100 mL Erlenmeyer flask, then add 5 mL concentrated HCl. Safety Precautions: The most common chemical exposure in this type of laboratory is a chemical splash to the skin or eye. Skin, mucous membranes, or eyes which have been splashed with commonly used chemicals or biologicals should be thoroughly washed for at least 15 minutes in cool tap water or eye wash station. Refer to MSDSs for additional chemical information. Report the incident immediately to a supervisor. Seek medical attention as necessary. n-Butyl chloride is flammable. Use in a vent hood or in a well ventilated area. Avoid contact with skin. Sodium hydroxide and hydrochloric acid are corrosive. Avoid contact with skin, eyes, and mucous membranes. Methanol is a flammable solvent. Use in a hood or in a well ventilated area. Chloroform is a chlorinated solvent and is considered a human carcinogen. Use in a hood or in a well ventilated area. Avoid contact with skin. Internal Standard (I.S.) Solution Preparation: Internal Standard Amount of Chemical Final Volume Solvent 0.5 mg/mL phenanthrene 1.00 g phenanthrene or 2.0 L n-butyl chloride or naphthalene naphthalene Transfer 1.00 g phenanthrene or naphthalene to a 2.0 L volumetric flask, and bring to volume with solvent. Mix by inverting flask. Calibration Standards: All solutions of stocks and standards are kept in the refrigerator. Allow the solutions to come to room temperature before using them. The concentration, amount, and volume of the calibration stock and calibration standards may vary proportionally depending on the availability of drug and the expected concentration of the drug. The following is provided as a guide: Calibration Stock 5.0 mg/mL Drug Amount of Drug (as base) 50.0 mg Final Volume 10 mL
3
Solvent deionized water

Transfer 50.0 mg drug calculated as the free base to a 10 mL volumetric flask, and bring to volume with deionized (DI) water. Mix by inverting flask. Sonicate as needed. Transfer to a 15 mL screw cap culture tube for storage. Calibration Procedure: 1. Choose a minimum of three appropriate calibration standards for the calibration curve based on the linear range required for the sample or drug to be analyzed. 2. Using a Hamilton syringe and/or volumetric pipette, place the appropriate amount of calibration stock into a 15 mL screw cap culture tube. Add deionized (DI) water to make a total volume of 5.0 mL. Calibration Standards 0.10 mg/mL 1.0 mg/mL 5.0 mg/mL 3. 4. 5. 6. Amount of Calibration Stock 0.10 mL 1.0 mL 5.0 mL Amount of DI H2O 4.90 mL 4.0 mL 0 mL Volume I.S. solution 5.0 mL 5.0 mL 5.0 mL
Add 5 drops of 40% sodium hydroxide and 5.0 mL internal standard solution. Cap the tubes and place on a rotator for 5 - 10 minutes. Centrifuge for 5 - 10 minutes on medium setting. Transfer an aliquot of the organic layer to an autosampler vial. Note: n-butyl chloride will be on the top; chloroform on the bottom. 7. Inject the extracted calibration standards into the GC using the appropriate instrument method. 8. Determine if the data are acceptable using the instructions found in the Calibration Curve/Response Factor Log. An acceptable calibration curve has a correlation coefficient, r2 > 0.995. If the curve is acceptable, file it in the Calibration Curve/Response Factor Log located in the Drug Laboratory. If not, seek supervisory assistance as needed. Quality Control Standard: Ideally the quality control standard is made from a different lot number or manufacturer than the calibration stock standard. At a minimum, it should be made from a different stock solution. A quality control standard must be successfully run on each day this procedure is used. The concentration, amount, and volume may vary proportionally depending on the availability of drug. The following is provided as a guide: QC Stock Amount of Drug (as base) Final Volume Solvent 2.0 mg/mL Drug 50 mg 25 mL deionized water Transfer 50.0 mg drug calculated as the free base to a 25 mL volumetric flask, and bring to volume with deionized water. Mix by inverting flask. Sonicate as needed. Transfer to a 25 mL screw cap culture tube for storage.
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory Alkaline Drug Analysis Version 2.0
Quality Control Procedure: 1. Using a volumetric pipette, place 2.5 mL of quality control stock and 2.5 mL DI water into a 15 mL labeled, screw cap culture tube. Add 5 drops of 40% sodium hydroxide and 5.0 mL internal standard solution. QC Standard 1.0 mg/mL Drug Amount of QC Stock Amount of DI H2O 2.5 mL 2.5 mL Volume of I.S. solution 5.0 mL
2. Cap the tube and place on rotator for 5 - 10 minutes. 3. Centrifuge for 5 - 10 minutes on medium setting. 4. Transfer an aliquot of the organic layer to an autosampler vial. Note: n-butyl chloride will be on the top and chloroform on the bottom of the respective water layers. 5. Inject the extracted quality control sample into the GC using the appropriate instrument method. Evaluation Criteria: 1. If the QC sample is within the set limits of +/- 5% of the target concentration, proceed with analysis. 2. If the QC sample is not within the set limits of +/- 5% of the target concentration, additional action must be taken to obtain an acceptable QC result such as re-running QC, re-extracting and re-running QC, making new stock with re-extraction and re-analysis, recalibrating (run curve), and/or consulting a supervisor. Color Testing: See the section on Color Testing. Spot tests will usually be run on each item of non-pharmaceutical evidence within one case up to the applicable statutory weight limit. Color testing may be performed on pharmaceutical evidence as applicable. Analytical Procedure: 1) Powdered Samples, Hard Chunky Powders, and Other Solids: a) Obtain the total weight of the material in the exhibit to be analyzed and record on the Drug Protocol Worksheet. b) Create a representative specimen by taking a sample from many or all of the solid pieces in the exhibit. If powder is also present, remove a sample and add to the material scraped from the solid pieces. Pulverize the resulting composite sample as necessary and analyze as described. c) GC/MS Identification i) Transfer approximately 10 mg of material to approximately 1 mL methanol in a test tube. (1) The analyst will vary the amount of material and methanol as needed to obtain an acceptable mass spectrum. ii) Vortex and centrifuge as necessary.
iii) Transfer an aliquot to an autosampler vial and inject on the GC/MS using BLANK.M method. d) GC Quantitation i) Weigh approximately 10 - 50 mg of the material to be analyzed, and record the weight on the Drug Protocol Worksheet. ii) Transfer sample to a 15 mL labeled screw cap culture tube. iii) Add 1.0 - 5.0 mL of deionized water using a volumetric pipette and 1 - 5 drops of 40% NaOH using a Pasteur pipette to the culture tube. iv) Add 1.0 - 5.0 mL internal standard solution. v) Cap the tube and place on rotator for 5 - 10 minutes. vi) Centrifuge for 5-10 minutes on medium setting. vii) Transfer an aliquot of the organic layer to an autosampler vial. Note: n-butyl chloride will be on the top; chloroform on the bottom. viii) Inject extracted sample into GC using appropriate instrument method and calculate the results. 2) Time Release Resin Beads a) Obtain the total weight of the material in the exhibit to be analyzed and record on the Drug Protocol Worksheet. b) GC Quantitation i) Weigh approximately 10 - 50 mg of the material to be analyzed and record the weight on the Drug Protocol Worksheet. ii) Transfer sample to a 15 mL labeled screw cap culture tube. iii) Add 1.0 - 5.0 mL of 25% HCl. Allow to stand overnight. iv) Add 1 - 5 drops of 40% NaOH, vortex, and check pH using pH paper. The solution should be alkaline. If not add 40% NaOH dropwise until the solution is alkaline. v) Add 1.0 - 5.0 mL of internal standard solution vi) Cap the tube and place on rotator for 5 - 10 minutes. vii) Centrifuge for 5-10 minutes on medium setting. viii) Transfer an aliquot of the organic layer to an autosampler vial. Note: n-butyl chloride will be on the top; chloroform on the bottom. ix) Inject extracted sample into GC using appropriate instrument method and calculate the results. c) GC/MS Identification i) Analyze a portion of the extract by GC/MS using BLANK.M method. 3) Tablets, Capsules, and Other Solid Pharmaceutical Preparations: a) Obtain the total weight of the material in the exhibit to be analyzed and record on the Drug Protocol Worksheet. b) Create a representative sample for analysis and grind to a powder if necessary. i) Capsules typically a composite sample is used for analysis. ii) Tablets typically one tablet is selected for analysis. c) Logo Identification i) Refer to Pharmaceutical Analysis Procedure d) GC/MS Identification i) Transfer an amount of sample that is equivalent to approximately 1 mg of drug to
approximately 1 mL methanol in a test tube. (1) The analyst will vary the amount of material and methanol as needed to obtain an acceptable mass spectrum. ii) Vortex and centrifuge as necessary. iii) Transfer an aliquot to an autosampler vial and inject on the GC/MS using BLANK.M method. e) GC Quantitation i) Weigh approximately 10 - 50 mg of the material to be analyzed, and record the weight on the Drug Protocol Worksheet. ii) Transfer sample to a 15 mL labeled screw cap culture tube. iii) Add 1.0 - 5.0 mL of deionized water using a volumetric pipette and 1 - 5 drops of 40% NaOH using a Pasteur pipette to the culture tube. iv) Add 1.0 - 5.0 mL internal standard solution. v) Cap the tube and place on rotator for 5 - 10 minutes. vi) Centrifuge for 5-10 minutes on medium setting. vii) Transfer an aliquot of the organic layer to an autosampler vial. Note: n- butyl chloride will be on the top; chloroform on the bottom. viii) Inject extracted sample into GC using appropriate instrument method and calculate the results. 4) Liquid Samples: a) Liquid samples may be analyzed by weight or by volume. If the sample is to be analyzed by volume, a known volume of the liquid must be weighed to determine density. i) If the volume of liquid is small, it should be treated as a residue; see below. ii) Obtain the total weight of liquid in the exhibit to be analyzed and record on the Drug Protocol Worksheet. (1) If the liquid is to be analyzed by volume, record the total volume on the Drug Protocol Worksheet. iii) Mix the liquid well before sampling. iv) GC/MS Identification (1) Place approximately 1 part sample to 4 parts methanol in a 15 mL screw cap culture tube. (a) The analyst will vary the amount of material and methanol as needed to obtain an acceptable mass spectrum. (2) Transfer an aliquot to an autosampler vial. Analyze by GC/MS using BLANK.M method. v) GC Quantitation (1) Transfer a known volume or weight of the sample to a 15 mL labeled screw cap culture tube. (2) If the liquid is aqueous, proceed to step 3. (a) If the composition of the liquid is unknown or organic, add three drops of methanolic HCl and evaporate the sample to dryness or to constant volume. (3) Based on the estimated drug concentration of the material, determine the extraction volume. The volumes of water and extraction solvent should be equal: (a) Add 1.0 5.0 mL of deionized water, 1-5 drops of 40% NaOH, and 1.0 - 5.0 mL internal standard solution.
(b) Cap the tube and place on rotator for 5 - 10 minutes. (c) Centrifuge for 5 -10 minutes on medium setting. (d) Transfer an aliquot of the organic layer to an autosampler vial. Note: nbutylchloride will be on the top and chloroform on the bottom. (e) Inject the extracted sample into GC using appropriate instrument method and calculate the results. 5) Residue Samples: a) Carefully rinse the residue into a container using methanol. b) GC/MS Identification i) Run a methanol blank before each residue sample using DRUGANA.M or BLANK.M methods and place in case file. ii) Place an aliquot of the methanol washing into an autosampler vial. Analyze by GC/MS using BLANK.M method. c) GC Quantitation i) Transfer the sample/methanol mixture to a 15 mL glass screw cap culture tube, add 1-3 drops of methanolic HCl, and evaporate to dryness. ii) Based on the estimated drug concentration of the material, determine the extraction volume. Add the same amount of water and extraction solvent: (1) Add 1.0 5.0 mL of deionized water to residue, 1-5 drops of 40% NaOH, and 1.05.0 mL internal standard solution. iii) Cap the tube and place on a rotator for 5 - 10 minutes. iv) Centrifuge for 5 - 10 minutes on medium setting. v) Run a methanol blank before each residue sample using the same method used for the sample and place in case file. vi) Transfer an aliquot of the organic layer to an autosampler vial. Note: n-butyl chloride will be on the top and chloroform on the bottom. vii) Inject the extracted sample into GC using the appropriate instrument method and calculate the results. (1) If it is necessary to dilute the residue, return to the original washing and dilute. viii) When analysis is complete return any extracted sample remaining in the autosampler vial to the original culture tube used for the extraction. (1) Remove and discard the aqueous layer from the culture tube. (2) Add 1-3 drops of methanolic HCl to the extracted sample to convert the drug to its more stable hydrochloride salt form. (3) Take the extracted sample to residue, in a laboratory fume hood. Close with a screw cap. (4) Label the tube added by laboratory, include FL# and initials, and place this tube in the evidence bag along with the evidence. 6) Clandestine Methamphetamine Laboratory Samples: a) Determine whether the liquid sample is aqueous or organic. i) If the sample is a solid, discuss analysis with supervisor. b) Determine and record pH on Drug Protocol Worksheet. i) A pH of 1 indicates the presence of concentrated acid. In this case, no further analysis is done.
c) Obtain the total volume and weight of liquid in the exhibit to be analyzed and record on the Drug Protocol Worksheet. i) Calculate the density and record on the Drug Protocol Worksheet. d) Perform color tests and record results on Drug Protocol Worksheet. i) A positive sodium nitroprusside test is a good indication that methamphetamine is present. ii) For positive FeCl3 test follow procedure for analysis of GHB. e) Organic Samples i) Transfer 3 mL by volumetric pipette to a 15 mL screw cap tube and record volume and weight on Drug Protocol Worksheet. ii) Add three drops of methanolic HCl and evaporate to dryness. iii) Add approximately 2 mL of deionized water. f) Aqueous Samples i) Transfer 3 mL by volumetric pipette to a 15 mL screw cap tube and record volume and weight on the Drug Protocol Worksheet. g) To either organic or aqueous samples, add four drops 40% NaOH and 2.0 mL internal standard solution to the 15 mL screw cap tube. h) Cap the tube and place on a rotator for 5 - 10 minutes. i) Centrifuge for 5 - 10 minutes on medium setting. j) Transfer an aliquot of the organic layer to an autosampler vial. k) GC/MS Identification i) Inject the extracted sample into the GC/MS using BLANK.M method. ii) LOWDOSE.M method may also be required for dilute samples. iii) If GC/MS results are negative, report as no controlled substance. (1) Record the presence of precursors/byproducts such as ephedrine/pseudoephedrine, triprolidine, etc. l) GC Quantitation i) If GC/MS results are positive for methamphetamine, inject the extracted sample into GC using METHAMPHETAMINE.M method and calculate the results. ii) Report the weight of the methamphetamine and the percent of methamphetamine in the sample. Report the volume of the liquid below the total weight. m) If the volume of liquid in the original container has been provided, calculate and report the total weight based on the analytical results of the sample vial. n) If the dimensions of the original container are provided, see below to calculate volume of liquid: cylinder: elliptical cylinder: V = pi r2 h , where r = radius in cm and h = height of liquid in cm. Volume units are in cm3 which is the same as mL. V = pi (ab)/4 h, where a = width of base in cm; b = length of base in cm; and h = height of the liquid in cm. Volume units are in cm3 which is the same as mL. V = abh, where a = width of base in cm; b = length of base in cm; and h = height of the liquid in cm. Volume units are in cm3 which is the same as mL.
rectangle or cube:
Instrument Parameters:
See GC and GC/MS Methods Notebooks in Drug Laboratory. Reporting Criteria: 1. Report both the quantity and the identity of the drug when the concentration of the drug is within +/- 20% of the calibration curve on the GC and the GC/MS is complete. It may be necessary to dilute extracted samples to bring them within the curve range. 2. Report the amount of the drug as insufficient for quantitation when the drug concentration is below 20% of the calibration curve on the GC and the GC/MS is complete. Calculations: 1. Dilutions a. In any situation in which a dilution is made, multiply the concentration of drug (mg/mL) by the dilution factor. A notation relating to how dilutions were made must be made in the case file. 2. Materials Analyzed by Volume: a. concentration of drug (mg/mL) x volume of internal standard solution used for extraction (mL) x total exhibit volume/extracted sample volume = mg of drug in the exhibit as the free base 3. Materials Analyzed by Weight: a. concentration of drug (mg/mL) x volume of internal standard solution used for extraction (mL) x total exhibit weight (mg)/extracted sample weight (mg) = mg of drug in the exhibit as the free base 4. Percent drug in the exhibit: a. mg of drug in the exhibit/total weight of the exhibit x 100 = % of drug in the exhibit 5. Amount drug in mg/100 mL (typically for codeine cough syrup): a. mg / 100 mL = mg (drug) X 100 Total Volume of exhibit
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Training Notes: Alkaline Procedure 1. If GC/MS results are negative when run on BLANK.M, run on alternative GC/MS methods such as LOWDOSE.M, LSD.M, etc. until satisfied that no controlled substance is present. 2. If a substance is dissolved in methanol and is determined to contain acetaminophen by GC/MS, the material should be extracted using this alkaline procedure and analyzed by GC and GC/MS to determine if codeine or hydrocodone is present. 3. GC quantitation of hydrocodone must be performed using chloroform - not n-butyl chloride as extracting solvent. 4. A sonicator may be used to bring materials into solution. Care should be used regarding the length of time materials are left in the sonicator since heat may decompose some drugs. Typically, a sonication time of 5 minutes or less is used. 5. Once a quantitation procedure is begun, the extraction procedure should be followed through to completion and not stopped mid-extraction. Prolonged exposure of drugs to a basic solution may cause decomposition. 6. The choice of internal standard - phenanthrene or naphthalene - is made to avoid coelution of the internal standard with the drug. 7. The toxicity of n-butyl chloride is less than that of chloroform; therefore, where possible, nbutyl chloride is the extraction solvent of choice. However, extraction efficiency of some drugs is low or variable in this solvent, and in this case, chloroform is used. 8. Dihydrocodeinone is an alternate name for hydrocodone. It is used to identify hydrocodone preparations listed in Penalty Group 3. Report as hydrocodone (dihydrocodeinone). 9. If the Marquis color test turns green, the extracted sample must be analyzed by GC/MS in order to determine if 4-bromo 2, 5-dimethoxyphenethylamine (Polo) is present. 10. Diphenhydramine and ketamine co-elute when run by GCMS using BLANK.M. Use SLOWRAMP.M method if there is evidence of the mixture. 11. TFMPP and MDMA may co-elute when run by GC/MS using BLANK.M. It may be necessary to use SLOWRAMP.M to resolve these peaks. 12. Depending on the solubility of the drug standard in water (salt/base form), the calibration stock may require a different solvent such as ethanol. Refer to the Merck Index or Clarkes Analysis of Drugs and Poisons for solubility information. For non-aqueous solutions take to residue before proceeding to extraction. 13. Calculation of drug weight as the free base: Option 1 To calculate the amount of drug as a salt required to produce a target amount of free base, use the following equation: MW drug (salt) / MW drug (free base) X target amount of free base Example: Amphetamine sulfate (C9H13N)2 H2SO4 MW = 368.5 Amphetamine (base) MW = 135.2 368.5 / 2 X 50 mg = 68.1 mg amphetamine sulfate 135.2 Note: Occasionally two molecules of drug are included in the salt form. Check the chemical formula prior to making standard. In this case, the molecular weight of the salt should be divided by 2 prior to using the formula above.
14. Calculation of drug weight as the free base: Option 2 To calculate the percentage of free base in the salt molecule to produce a target amount, use the following equation: Example: Amphetamine sulfate (C9H13N)2 H2SO4 MW = 368.5 Amphetamine (base) MW = 135.2 135.2 X 2 % base in salt = 368.5
=
0.734
Amount of salt needed = 50 mg = 68.1 mg amphetamine sulfate 0.734
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COCAINE DIRECT DILUTION COMBINED QUANTITATIVE AND QUALITATIVE ANALYSIS Principle of Assay: This method describes a direct dilution procedure for the identification and quantitation of cocaine in suspected drug samples. Cocaine is simultaneously identified and quantitated using gas chromatography/mass spectrometry (GC/MS). In this method, a known amount of suspected drug substance is quantitatively diluted in dichloromethane containing phenanthrene as the internal standard and analyzed by GC/MS. Sample Requirements: Powders and solids are suitable for analysis. Residues and other specimens suspected of containing cocaine should be analyzed using the Weak Alkaline Drug Analysis. Equipment: Gas chromatograph-mass spectrometer with split/splitless injector and mass selective detector containing a methylsiloxane capillary column such as an HP-5MS, 30 m x 0.25 mm x 0.25 um film thickness (GC/MS Methods Notebook in Drug Laboratory). Screw top culture tubes, 25 mL, 20 mm x 125 mm, screw cap with Teflon liner Screw top culture tubes, 15 mL, 16 mm x 125 mm, screw cap with Teflon liner Volumetric flask, 2 mL, 10 mL, 6.0 L Hamilton syringes, 100 uL, 250 uL, 500 uL Autosampler vials, 32 mm x 11 mm, Teflon lined seals and glass inserts Vortex mixer Centrifuge Sonicator Rotator Analytical balance Adjustable 10 mL Dispensette Reagent Bottle Pipettes, various Mortar and Pestle Reagents: Phenanthrene, reagent grade or better Dichloromethane (methylene chloride), reagent grade or better Analytical standards
Safety Precautions:
Cocaine Direct Dilution Analysis Version 2.0
The most common type of chemical or biological exposure in this type of laboratory is a splash to the skin or eye. Skin, mucous membranes, or eyes which have been splashed with commonly used chemicals or biologicals should be thoroughly washed for at least 15 minutes in cool tap water or eye wash station. Refer to appropriate MSDSs for additional chemical information. Report the incident immediately to a supervisor. Seek medical attention as necessary. Dichloromethane is a chlorinated solvent and is considered a suspected human carcinogen. Use in a hood or in a well ventilated area. Avoid contact with skin. Internal Standard (I.S.) Solution Preparation: Internal Standard Amount of Chemical Final Volume Solvent
0.5 mg/mL 3.0 g phenanthrene 6.0 L dichloromethane phenanthrene Weigh 3.0 g phenanthrene, transfer to a 6.0 L volumetric flask, and bring to volume with solvent. Mix by inverting flask. Calibration Standards: All solutions of standards and stocks are kept in the refrigerator. Allow the standards to come to room temperature before using. The concentration, amount, and volume of the calibration stock and calibration standards may vary proportionally depending on the availability of drug and the expected concentration range of the drug. The following is provided as a guide: Calibration Stock Amount of Drug (as base) Final Volume Solvent 4.0 mg/mL cocaine 40.0 mg 10 mL I. S. solution Transfer 40.0 mg drug calculated as the free base to a 10 mL volumetric flask and bring to volume with I.S. solution. Mix by inverting flask. Sonicate as needed. Transfer solution to a 15 mL culture tube for storage. Calibration Procedure: 1. Choose four appropriate calibration standards for the calibration curve based on the linear range required for the sample or drug to be analyzed. 2. Using a Hamilton syringe and/or volumetric pipette, place the appropriate amount of calibration stock into a 2 mL volumetric flask. Add internal standard solution to make a total volume of 2.0 mL. Mix by inverting the flask.
Calibration Standards 0.20 mg/mL 0.50 mg/mL
Amount of Calibration Stock 0.10 mL 0.25 mL
Volume of I.S. solution 1.90 mL 1.75 mL

1.0 mg/mL 2.0 mg/mL
0.50 mL 1.00 mL
1.50 mL 1.00 mL
3. Transfer an aliquot of the organic layer to an autosampler vial. Inject the extracted standards into the GC/MS using COCQUANT.M method. 4. Determine if data are acceptable using the instructions found in the GC/MS Quantitation Methods Notebook. An acceptable calibration curve has a correlation coefficient, r2 > 0.995. If the curve is acceptable, file it in the GC/MS Quantitation Methods Notebook located in the Drug Laboratory. If not, seek supervisory assistance as needed. Quality Control Standard: Ideally the quality control standard is made from a different lot number or manufacturer than the calibration stock. At a minimum, it should be made from a different stock solution. A quality control standard must be successfully run on each day this procedure is used. The concentration, amount, and volume may vary proportionally depending on the availability of drug. The following is provided as a guide: QC Stock Amount of Drug (as base) Volume of I.S. solution 4.0 mg/mL cocaine 40.0 mg 10 mL Transfer 40.0 mg drug calculated as the free base to a 10 mL volumetric flask and bring to volume with I.S. solution. Mix by inverting flask. Sonicate as needed. Transfer solution to 15 mL culture tube for storage. Quality Control Procedure: 1. Place the appropriate amount of QC stock into a 2 mL volumetric flask using a Hamilton syringe and/or a volumetric pipette. Add internal standard solution to make a total volume of 2.0 mL. Mix by inverting. QC Standard 1.0 mg/mL cocaine Amount of QC Stock 0.50 mL Volume of I.S. solution 1.50 mL
2. Transfer an aliquot to an autosampler vial, inject into the GC/MS using COCQUANT.M method. Evaluation Criteria: 1. If the QC sample is within the set limits of +/- 10% of the target concentration, proceed with analysis. 2. If the QC sample is not within the set limits of +/- 10% of the target concentration, additional action must be taken to obtain an acceptable QC result such as re-running QC, preparing and running a new QC from the stock solution, making a new stock solution and new QC, recalibrating (run curve), and/or consulting a supervisor.
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory Cocaine Direct Dilution Analysis Version 2.0
Color Testing: See the section on Color Tests. Spot tests will typically be run on each item of nonpharmaceutical evidence within one exhibit up to the applicable statutory limit. The color test must be positive to proceed with this procedure. Analytical Procedure: 1. Obtain the total weight of material in the exhibit and record on the Drug Protocol Worksheet. 2. Create a representative specimen by taking a sample from many or all of the solid pieces in the exhibit. Pulverize the resulting composite sample and analyze as described. 3. Weigh approximately 10-25 mg of material to be analyzed; record the weight on the Drug Protocol Worksheet. 4. Transfer the sample to a clean 15 mL labeled screw top culture tube. 5. Using a Dispensette Reagent Bottle, add 10 mL of internal standard solution and cap. 6. Rotate 5 - 10 minutes, vortex or sonicate as needed, and centrifuge for 5 - 10 minutes on medium setting. 7. Transfer an aliquot to a labeled autosampler vial and cap. 8. Inject the sample into the GC/MS using the COCQUANT.M method for quantitative/qualitative analysis, and calculate the results. Instrument Parameters: See GC/MS Methods Notebook located in Drug Laboratory. Reporting Criteria: 1. The reporting range of the assay is +/-20% of the calibration curve. 2. When the concentration of the drug is within the reporting range of the assay, the retention time of the sample is within 0.03 minutes of the quality control sample, and the GC/MS is complete, report both the quantity and the identity of the drug. 3. When the concentration of the drug is below the reporting range of the assay, the retention time of the drug is not within range, or the GC/MS is incomplete, reanalyze using the Weak Alkaline Drug Analysis. 4. If GC/MS is complete but below quantitation limit, reanalyze using the Weak Alkaline Drug Analysis. 5. If no indication of cocaine is present, the material is considered negative for cocaine; however, a sample of the material should be diluted with methanol and injected onto the GC/MS as an unknown using the BLANK.M method. Calculations: 1. Dilutions a. In any situation in which a dilution is made, multiply the concentration of drug
(mg/mL) by the dilution factor. A notation relating to how dilutions were made must be made in the case file. 2. Materials Analyzed by Weight a. concentration of drug (mg/mL) x volume of internal standard solution used for extraction (mL) x total exhibit weight (mg)/extracted sample weight (mg) = mg of drug in the exhibit as the free base 3. Percent drug in the exhibit: a. mg of drug in the exhibit/total weight of the exhibit x 100 = % of the drug in the exhibit
Training Notes: Cocaine Direct Dilution 1. If GC/MS results are negative when run on BLANK.M, run on alternative GC/MS methods such as LOWDOSE.M, LSD.M, etc. until satisfied that no controlled substance is present. 2. A sonicator may be used to bring materials into solution. Care should be used regarding the length of time materials are left in the sonicator since heat may decompose some drugs. Typically, a sonication time of 5 minutes or less is used. 3. For all drugs quantitated by GC/MS, acceptable calibration curves must be documented in the GC/MS Quantitation Methods Notebook. 4. The daily QC value, internal standard retention time, and drug retention time will be noted on the Drug Protocol Worksheet. 5. GC methodology may be used at any time or in conjunction with this quantitation procedure. Only the MS quantitation will be charged to the customer. 6. Care should be taken in the dispensing of 10 mL of dichloromethane. This is a volatile liquid and use of the dispensette should be slow and steady. 7. Federal cases containing cocaine require salt/base determination. Refer to the Color Testing procedure for presumptive salt/base determination and to the Infrared Spectroscopy procedure for conclusive identification of salt/base form. 8. Calculation of drug as free base: Option 1 To calculate the amount of drug as a salt required to produce a target amount of free base, use the following equation: MW drug (salt) / MW drug (free base) X target amount of free base Note: Occasionally two molecules of drug are included in the salt form, for example morphine sulfate. Check the chemical formula prior to making the standard. In the case of morphine sulfate, the molecular weight of the salt should be divided by 2 prior to using the formula above. Example: Cocaine HCl (C17H21NO4) HCl Cocaine (base) MW= 339.8 MW= 303.4
339.8 X 40 mg = 44.7 mg Cocaine HCl 303.4
9. Calculation of drug as free base: Option 2 Alternatively, to calculate the percentage of free base in the salt molecule to produce a target amount, use the following equation: Example: Cocaine HCl (C17H21NO4) HCl MW= 339.8 Cocaine (base) MW= 303.4 % base in salt = 303.4 = 0.893 339.8
Amount of salt needed = 40.0 mg = 44.7 mg cocaine HCl 0.893
COLOR TESTING PROCEDURE Principle of Assay: Color tests are used as a screening test at the beginning of an analysis. These color tests provide suggestive or tentative identification of chemical identity; however, they do not stand alone as conclusive identification of drug identity. Most are performed on clean porcelain or disposable plastic spot plates; however some may be performed in disposable culture tubes. The test reagent should be added to the plate or tube first, and then the questioned sample. This practice determines if the plate or tube was clean before the analysis. The color observed for each reagent used is recorded on the Drug Protocol Worksheet. Negative reactions may be recorded as negative or no reaction. Sample Requirements: Powders, solids, liquids, tablets, capsules, residues, and other types of drug evidence are suitable for analysis using this procedure. Equipment: Spot plates Spatula Test tubes Color test reagents as noted below Safety Precautions: Most color test reagents are comprised of strong acids and chemicals requiring careful handling. Appropriate safety precautions should be observed. Refer to MSDSs. The shelf life of color test reagents is two years. Reagents: 1) Cobalt Thiocyanate (CoSCN) Reagent (Ruybal) a) Reagent Preparation: i) 2% CoSCN solution (1) Place 12.6 grams CoSCN (crushed) in a graduated cylinder; bring to 630 mL with distilled H2O. ii) Concentrated H3PO4, 210 ml iii) Platinum chloride in 25% H3PO4 (1) 25% H3PO4 (a) Add 17.5 mL H3PO4 to 52.5 mL deionized water (2) Mix 3.5 grams platinum chloride (H2PtCl6 6 H2O) in 70 mL 25% H3PO4. b) Procedure: i) Mix by volume:
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory Color Testing Procedure Version 2.0
9 parts 2% CoSCN solution (630 mL) 3 parts concentrated H3PO4 (210 mL) 1 part platinum chloride solution (70 mL) Add CoSCN solution and H3PO4 to the platinum chloride solution. (Always add acid to H2O.) ii) Let stand one to two days. iii) Color will change from orange-yellow to pink. c) Results: i) Substances reported to produce a blue precipitate with this color test include cocaine (base and HCl), lidocaine, diphenhydramine, propylhexadrine, Triton X (surfactant), amitriptyline, dibucaine, mepivicaine, methadone, and PCP. 2) Determination of the Chemical Form of Cocaine a) Purpose: i) This procedure determines the chemical form of cocaine (base vs. salt) in the sample utilizing the differential solubility properties of the two compounds. b) Procedure: i) Place 10-15 mg of sample into a test tube and add approximately 1 mL of hexane. ii) Vortex the sample and allow particulates to settle; centrifuge if necessary. iii) Transfer 4-5 drops of the hexane solution to a clean spot test plate well containing 1-2 drops of cobalt thiocyanate solution, shake briefly. iv) The appearance of a characteristic blue indicates the presence of free base cocaine. v) To the original test tube, add approximately 1 mL of deionized water, vortex, and allow particulates to settle; centrifuge as necessary vi) Transfer 4-5 drops of the aqueous layer (bottom layer) to a clean spot test plate well containing 1-2 drops of cobalt thiocyanate solution, add 1-2 drops of chloroform, and shake briefly. vii) The appearance of a blue color in the chloroform layer indicates the presence of cocaine salt. viii) If both tests are negative, transfer 4-5 drops of the remaining hexane solution to a clean spot test plate well containing 1-2 drops of cobalt thiocyanate solution, add 1-2 drops of chloroform, and shake briefly. A positive result indicates that the original sample is a mixture of cocaine HCl and sodium bicarbonate. A negative result confirms the absence of any cocaine in the sample. Hexane blue blue no color change no color change H2O + CHCl3 blue no color change blue no color change Cocaine_Present Yes Yes Yes No Chemical Form Salt + Base Base Salt
(1) (2) (3) (4)
3) Marquis Reagent a) Reagent Preparation: i) Add 13 15 drops of 37% formaldehyde solution to 15 mL concentrated H2SO4. b) Results:
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory 2 Color Testing Procedure Version 2.0
i) Color changes are as follows: Drug Color change Amphetamines orange to brown Opiates violet Diphenhydramine yellow Amitriptyline red to brown MDA, MDMA, MDEA, N-OH MDA purple to black Aspirin pink to cherry red upon standing 4) Mecke Reagent a) Reagent Preparation: i) Dissolve 250 mg selenious acid in 25 mL concentrated sulfuric acid. ii) Results: (1) Color changes are as follows: Drug Opiates Diphenhydramine MDA, MDMA, MDEA, N-hydroxy MDA Color Change green yellow deep aqua blue to black
5) Sodium Nitroprusside/Acetaldehyde a) Reagent Preparation: i) Solution A: (1) Make a 1% solution of sodium nitroprusside (sodium nitroferricyanide) in water. (2) Add 10% by volume of acetaldehyde. (3) Example: Place 0.9 gram of sodium nitroprusside in 90 mL H2O. Mix thoroughly. In a hood, add 10 mL acetaldehyde; watch for bumping. Mix. (4) The stock is refrigerated when not in use. ii) Solution B: (1) Make a 2% sodium carbonate solution in water. (2) Example: Place 2 grams sodium carbonate in 100 mL H2O. Mix. b) Procedure: i) Place a small amount of powder in a spot plate. ii) Add one drop of Solution A followed by two drops of Solution B. iii) An immediate blue color indicates presence of a secondary amine such as methamphetamine. Amphetamine and other primary amines yield a slow pink to cherry red color. c) Results: i) Color changes are as follows: Drug Methamphetamine Amphetamine MDEA, MDMA MDA
Color Change blue slow pink to cherry red blue negative

Color Testing Procedure Version 2.0
6) Ferrous Sulfate for Nitroglycerine a) Reagent Preparation: i) To one volume of a 10% solution of ferrous sulfate (FeSO4.7H2O), add 5 volumes of sulfuric acid with cooling. ii) For example, mix 0.1 g FeSO4.7H2O and 1 mL distilled water; add 5 mL H2SO4. b) Results: i) A red or pink color indicates the presence of nitroglycerine. 7) Silver Nitrate Test for the Presence of Chloride Ion a) Reagent Preparation: i) 5% silver nitrate (1) Dissove 5g silver nitrate in 100 mL deionized water. (2) Mix well. b) Procedure: i) Place powder in test tube and dissolve in distilled water. ii) Add a few drops of 5% silver nitrate and shake. iii) A precipitate will form if chloride ions are present. iv) Add a few drops of concentrated nitric acid and shake. c) Results: i) If precipitate persists, the test is positive for the presence of chloride ions. 8) Van Urk Reagent for Hallucinogens a) Reagent Preparation: i) Dissolve 2 grams of para-dimethylaminobenzaldehyde in 50 mL of ethanol ii) Add 50 mL of concentrated hydrochloric acid. b) Results: i) Slow development of lavender color indicates presence of LSD, LAMPA, psilocybin, psilocin, etc. 9) Ferric Chloride Test for GHB a) Reagent Preparation: i) Dissolve 5 grams ferric chloride in 100 ml deionized water. b) Results: i) An orange color is consistent with the presence of gamma-hydroxybutyrate (GHB). ii) Gamma-butyrolactone (GBL) and 1,4 butanediol do not produce a color change. `` 10) Duquenois-Levine for Marihuana Reagent Preparation: a) Reagent Preparation: i) Mix 1 gram vanillin, 50 mL ethanol, and mL acetaldehyde. b) Procedure and Results: i) Refer to the Marihuana Procedure. Color Test Procedure: To a clean spot test well, add 1 2 drops reagent; then add a small amount of material to be tested.
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory Color Testing Procedure Version 2.0
Reporting Criteria: 1) Describe color changes or lack of change on the Protocol Sheet. Quality Control Procedure Stock solutions of color test reagents will be made up as needed. They may be prepared in bulk volume and distributed among analysts. After they are made, they will be checked by the person preparing the solution as well as checked by each analyst before put into use. The color reagents will be checked with the Color Reagent Test Kit. The test kit contains a variety of drugs necessary to test the color test reagents including cocaine, MDMA, methamphetamine, PCP, GHB, ephedrine, and marihuana. Materials in the test kit have been removed from cases or have been purchased and have been verified by GC/MS and or IR. Common Reagents and Appropriate Check Compounds Reagent Duquenois-Levine Marquis Mecke CoSCN Van Urk FeCl3 Nitroprusside Negative Control Check Compound Marihuana Heroin, MDMA, Methamphetamine Heroin, MDMA Cocaine, PCP LSD GHB Methamphetamine, MDMA Ephedrine
Procedure: 1. At the beginning of each month all routine color testing reagents (Duquenois, Marquis, Mecke, CoSCN, Nitroprusside) will be checked with a positive and a negative control using the Color Reagent Test Kit. 2. The results of the color change will be documented on the Monthly Color Reagent Test Log by the analyst(s) conducting the test(s). 3. If there is no change in color document NR or no reaction. 4. If a reagent does not produce the appropriate and expected color change, a second known standard will be evaluated with the same solution. If this second test also produces a negative response, all reagent bottles with this solution will be collected, discarded, and fresh solution prepared. 5. Non routine color reagents (Van Urk, FeCl3,) will be checked for validity before use.
SELECTED COLOR TEST OBSERVATIONS A blank entry indicates no color change Individual Drugs Cobalt Thiocyanide Sodium Nitroprusside
Drug Acetaminophen Asprin 4-Bromo-2,5dimethoxyphenethylamine (Polo) Dextromethorphan Doxepin Ephedrine SO4, Ephedrine HCl Fluoxetine (Prozac) Guaifenesin MBDB MDA MDA, isopropenyl MDA, N-hydroxy MDEA MDMA Methylphenidate N-Methylcathinone Phenethylamine Phentermine Propylhexedrine Tramadol
Marquis pink to deep red bright green
Mecke
blue specks blue
slow black maroon
yellow-red
deep pink
slow blue, diffuse
orange-red purple deep purple deep purple purple, sparse purple deep purple deep purple
deep pink slow blue pink bright blue
few blue specks few blue specks
deep aqua deep aqua blue purple deep aqua deep aqua deep aqua
bright blue bright blue
blue orange orange orange-brown orange deep green, slow blue
blue blue
SELECTED COLOR TEST OBSERVATIONS Drugs in Combination Cobalt Thiocyanide blue Sodium Nitroprusside
Drug Aspirin, Cocaine, and Guaifenesin Ephedrine, Caffeine, and Phenylpropylamine (tablets with green specks) Ephedrine, Cocaine, Guaifenesin Ephedrine and Dextromethorphan (speckled round tablet ) Ephedrine and Guaifenesin Methamphetamine, Guaifenesin, and Ephedrine Phenylpropanolamine and Dextromethorphan
Marquis purple
Mecke maroon
pink
blue
purple
green-purple
blue green specks blue specks
purple to black deep purple
pink-yellow
deep pink
deep aqua
deep purpleviolet blue-green specks purple-black
green-violet
blue
pink-yellow
deep pink
DIRECT DILUTION ANALYSIS Principle of Assay: This method describes a direct dilution procedure for a number of controlled substances including many opiates and benzodiazepines. Drugs are identified using analytical techniques which provide conclusive identification of the drug. These techniques include gas chromatography/mass spectrometry (GC/MS), and infrared spectroscopy (IR). Drugs are usually quantitated by gas chromatography (GC). A known amount of suspected drug substance is quantitatively diluted in ethanol containing phenanthrene as the internal standard. Specific details of quantitation for each drug and its linear range of quantitation may be found in the Calibration Curve/Response Factor Log for each GC. Examples of drugs analyzed using the direct dilution procedure follow: Alprazolam Heroin Oxazepam Clonazepam Lorazepam Prazepam Diazepam Monoacetylmorphine Temazepam Flunitrazepam Morphine Triazolam Flurazepam Olanzapine Marked pharmaceuticals will usually be identified by logo match and GC/MS without quantitation. However when the penalty group or schedule of a marked pharmaceutical preparation is determined by dosage or preparation, sufficient testing should be performed for placement of the substance in a proper penalty group or schedule. Sample Requirements: Powders, solids, tablets, capsules, residues, and plant extracts (for example, raw opium and morphine base) are suitable for analysis. Liquids are not usually suitable for analysis by this procedure. Equipment: Gas chromatograph-mass spectrometer containing a methylsiloxane capillary column such as an HP-5MS, 30 m x 0.25 mm x 0.25 um film thickness and gas chromatograph-flame ionization detector with split/splitless injector containing a methylsiloxane capillary column such as an HP1, 15 m x 0.32 mm x 1.0 um film thickness (see GC/MS and/or GC Methods Notebooks in Drug Laboratory). Screw top culture tubes, 25 mL, 20 mm x 125 mm, screw cap with Teflon liner Screw top culture tubes, 15 mL, 16 mm x 125 mm, screw cap with Teflon liner Volumetric flask, 5 mL, 10 mL, 25 mL, 500 mL Autosampler vials, 32 mm x 11 mm, Teflon lined seals and glass inserts Hamilton syringe, 250 uL Vortex mixer
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory Direct Dilution Analysis Version 2.0
Centrifuge Sonicator Test tubes Analytical balance Pipettes, various Graduated cylinders, various Adjustable 2 mL Dispensette Reagent Bottle Mortar and Pestle Erlenmeyer flask, 100 mL Reagents: Phenanthrene, reagent grade or better Methanol, reagent grade or better Ethyl Alcohol, USP grade, 200 proof - Absolute Chloroform, reagent grade or better HCl, reagent grade or better Sodium bicarbonate, reagent grade or better Analytical standards Methanolic HCl (5%) Transfer 5 mL concentrated HCl and 95 mL of methanol to a 100 mL Erlenmeyer flask. Safety Precautions: The most common type of chemical or biological exposure in this type of laboratory is a splash to the skin or eye. Skin, mucous membranes, or eyes which have been splashed with commonly used chemicals or biologicals should be thoroughly washed for at least 15 minutes in cool tap water or eye wash station. Refer to appropriate MSDSs for additional chemical information. Report the incident immediately to a supervisor. Seek medical attention as necessary. Methanol and ethanol are flammable solvents. Avoid contact with skin, eyes, and mucous membranes. Chloroform is a chlorinated solvent and is considered a human carcinogen. Use in a hood or in a well ventilated area. Avoid contact with skin, eyes, and mucous membranes. Hydrochloric Acid is corrosive. Avoid contact with skin, eyes, an mucous membranes. Internal Standard (I.S.) Solution Preparation: Internal Standard Amount of Chemical Final Volume Solvent
0.5 mg/mL 0.25 g phenanthrene 500 mL ethanol phenanthrene Weigh 0.25 g phenanthrene, transfer to a 500 mL volumetric flask, and bring to volume with solvent. Mix by inverting the flask.
Calibration Standards: All solutions of stocks and standards are kept in the refrigerator. Allow the standards to come to room temperature before using. The concentration, amount, and volume of the calibration stock and calibration standards may vary proportionally depending on the availability of drug and the expected concentration range of the drug. The following is provided as a guide. Calibration Stock Amount of Drug (as base) Volume of I.S solution
2.0 mg/mL Drug 20 mg 10 mL Transfer 20.0 mg drug calculated as the free base to a 10 mL volumetric flask and bring to volume with I.S. solution. Mix by inverting the flask. Sonicate as needed. Transfer solution to a 15 mL culture tube for storage. Calibration Procedure: 1. Choose a minimum of three appropriate calibration standards for the calibration curve based on the linear range required for the sample or drug to be analyzed. 2. Using a Hamilton syringe or a volumetric pipette, place the appropriate amount of calibration stock into a 5 mL volumetric flask. Add I.S. solution to make a total volume of 5.0 mL. Mix by inverting the flask. Sonicate as needed. Calibration Standards 0.10 mg/mL 1.0 mg/mL 2.0 mg/mL Amount of Calibration Stock 250 uL 2.5 mL 5.0 mL Volume I.S. solution Dilute to 5 mL Dilute to 5 mL 0 mL appropriate Calibration correlation Calibration supervisory
3. Transfer an aliquot to an autosampler vial. Inject into the GC using the instrument method. 4. Determine if the data are acceptable using the instructions found in the Curve/Response Factor Log. An acceptable calibration curve has a coefficient, r2 > 0.995. If the curve is acceptable, file it in the Curve/Response Factor Log located in the Drug Laboratory. If not, seek assistance as needed. Quality Control Standard:
Ideally the quality control standard is made from a different lot number or manufacturer than the calibration stock. At a minimum, it should be made from a different stock solution. A quality control standard must be successfully run on each day this procedure is used. The concentration, amount, and volume may vary proportionally depending on the availability of
drug. The following is provided as a guide: QC Stock Amount of Drug (as base) Volume of I.S. solution 1.0 mg/mL Drug 10 mg 10 mL Transfer 10.0 mg drug calculated as the free base to a 10 mL volumetric flask and bring to volume with internal standard (I.S.) solution. Mix by inverting the flask. Sonicate as needed. Transfer solution to a 15 mL culture tube for storage. Quality Control Procedure: 1. Transfer an aliquot of QC Stock to an autosampler vial, inject into GC using the appropriate instrument method. QC Standard 1.0 mg/mL Drug Evaluation Criteria: 1. If the QC sample is within the set limits of +/- 5% of the target concentration, proceed with analysis. 2. If the QC sample is not within the set limits of +/- 5% of the target concentration, additional action must be taken to obtain an acceptable QC result such re-running QC, reprepping and re-running the QC, making new stock solution and reanalyzing, recalibrating (run curve), and/or consulting a supervisor. Color Testing: See the section on Color Tests. Spot tests will typically be run on each item of nonpharmaceutical evidence within one exhibit up to the applicable statutory limit. Color testing may be performed on pharmaceutical evidence as applicable. Analytical Procedure: 1. Powder Samples and Other Solids: a. Obtain the total weight of the suspected drug material to be analyzed and record on the Drug Protocol Worksheet. b. Create a representative specimen by taking a sample from many or all of the solid pieces in the exhibit. If powder is also present, remove a sample and add to the material scraped from the solid pieces. Pulverize the resulting composite sample and analyze as described. c. GC/MS Identification i. Transfer approximately 10 mg of material to be analyzed to approximately 1 mL methanol in a test tube. 1. The analyst will vary the amount of material and methanol as needed to obtain an acceptable mass spectrum.
Amount of Drug (as base) Use QC Stock
Volume of I.S solution -----------
ii. Vortex and centrifuge as necessary. iii. Transfer an aliquot to an autosampler vial and inject on the GC/MS using BLANK.M method. d. GC Quantitation i. Weigh approximately 10 - 50 mg of the material to be analyzed, and record the weight on the Drug Protocol Worksheet. ii. Transfer sample to a 15 mL labeled screw cap culture tube. iii. Add 1.0 - 5.0 mL of internal standard solution depending upon the expected concentration of the drug; document the volume used on the Drug Protocol Worksheet. iv. Rotate for 5 - 10 minutes. Vortex or sonicate as needed. v. Centrifuge for 5-10 minutes on medium setting. vi. Transfer an aliquot to an autosampler vial, inject the sample into GC using the appropriate instrument method, and calculate the results. 2. Tablets, Capsules, and Other Pharmaceutical Preparations: a. Obtain the total weight of the suspected drug material to be analyzed and record on the Drug Protocol Worksheet. b. Create a representative sample for analysis: i. Capsules typically a composite sample is used for analysis. ii. Tablets typically one tablet is selected for analysis. c. Logo Identification i. Refer to Pharmaceutical Analysis Procedure. d. GC/MS Identification i. Transfer an amount of sample that is equivalent to approximately 1 mg of drug to approximately 1 mL methanol in a test tube. 1. The analyst will vary the amount of material and methanol as needed to obtain an acceptable mass spectrum. ii. Vortex and centrifuge as necessary. iii. Transfer an aliquot to an autosampler vial and inject on the GC/MS using BLANK.M method. e. GC Quantitation i. Weigh approximately 10 - 50 mg of material to be analyzed and record on the Drug Protocol Worksheet. ii. Transfer sample to a 15 mL labeled screw cap culture tube. iii. Add 1.0 - 5.0 mL of internal standard solution depending upon the expected concentration of the drug and record the volume on the Drug Protocol Worksheet. iv. Rotate for 5 - 10 minutes. Vortex and sonicate as needed. v. Centrifuge for 5-10 minutes on medium setting. vi. Transfer an aliquot to an autosampler vial, inject the sample into the GC using the appropriate instrumental method, and calculate the results. 3. Residue Samples: a. Carefully rinse the residue into a small container using methanol. Make a notation that the specimen is a residue on the Drug Protocol Worksheet.
b. GC/MS Identification i. Run a methanol blank before each residue sample using DRUGANA.M or BLANK.M methods and place in case file. ii. Place an aliquot of the methanol washing in an autosampler vial. iii. Vortex and centrifuge as necessary. iv. Inject on the GC/MS using BLANK.M method. c. GC Quantitation i. Place the sample/methanol mixture in a 15 mL labeled screw top culture tube, add 1-3 drops of methanolic HCl and evaporate to dryness. ii. Based on the estimated drug concentration of the material, determine the dilution volume. Add 1.0 - 5.0 mL of internal standard solution. iii. Sonicate for 5 - 10 minutes. iv. Centrifuge for 5-10 minutes on medium setting. v. Transfer an aliquot to an autosampler vial. vi. Run a methanol blank before each residue sample using the same method used for the sample and place in case file. vii. Inject the sample into the GC using the appropriate instrument method, and calculate the results. d. When analysis is complete return any sample remaining in the autosampler vial to original culture tube. e. Under a laboratory fume hood, add 1-3 drops of methanolic HCl to the sample and evaporate to dryness. f. Label the tube added by the laboratory, include FL#, initials, and place this tube in the evidence bag along with the evidence. 4. Low Dose Heroin Identification: a. When low dose heroin is cut with acetaminophen, GC/MS identification of heroin may be obscured by the acetaminophen. The following procedure separates acetaminophen from heroin and improves the GC/MS. Quantitation of heroin is performed using Procedure 1 above. Make a notation on the Drug Protocol Worksheet when this procedure is used. b. GC/MS Identification i. Dissolve 20-30 mg sample in approximately 1 mL of water in a test tube. 1. The analyst will vary the amount of material as needed to obtain an acceptable mass spectrum. ii. Add NaHCO3, and approximately 1mL of chloroform. iii. Centrifuge as necessary. iv. Transfer an aliquot of the chloroform layer (bottom) to an autosampler vial and inject the sample into the GC/MS using BLANK.M method. Instrument Parameters: See GC and GC/MS Methods located in Drug Laboratory. Reporting Criteria:
Direct Dilution Analysis Version 2.0
1. Report both the quantity and the identity of the drug when the concentration of the drug is within +/- 20% of the calibration curve on the GC and the GC/MS is complete. It may be necessary to dilute extracted samples to bring them within the curve range. 2. Report the amount of the drug as insufficient for quantitation when the drug concentration is below 20% of the calibration curve on the GC and the GC/MS is complete. Calculations: 1. Dilutions a. In any situation in which a dilution is made, multiply the concentration of drug (mg/mL) by the dilution factor. A notation relating to how dilutions were made must be made in the case file. 2. Materials Analyzed by Weight a. concentration of drug (mg/mL) x volume of internal standard solution used for extraction (mL) x total exhibit weight (mg)/extracted sample weight (mg) = mg of drug in the exhibit as the free base 3. Percent drug in the exhibit: a. mg of drug in the exhibit/total weight of the exhibit x 100 = % of the drug in the exhibit
Training Notes: Direct Dilution Analysis 1. If GC/MS results are negative when run on BLANK.M, run on alternative GC/MS methods such as LOWDOSE.M, LSD.M, etc. until satisfied that no controlled substance is present. 2. When analyzing a morphine sulfate tablet add approximately 100 mg of NaHCO3 to the internal standard solution before sonication. 3. A sonicator may be used to bring materials into solution. Care should be used regarding the length of time materials are left in the sonicator since heat may decompose some drugs. Typically, a sonication time of 5 minutes or less is used. 4. Liquid morphine exhibits are typically quantitated using the Morphine Syrup Analysis procedure. 5. The heroin monohydrate standard is most often purchased in dilutable form. It is acceptable to prepare and store the solution in the dilutable vial. 6. Identify material as opium when morphine plus four of the following alkaloids (codeine, noscapine, thebaine, papaverine, meconin) are present. Quantitation is not necessary. Report total weight of material. 7. Black tar heroin is frequently encountered as a sticky solid material. It may be necessary to freeze the sample for 5-10 minutes before proceeding with analysis. 8. Calculation of drug as free base: Option 1 To calculate the amount of drug as a salt required to produce a target amount of free base, use the following equation: MW drug (salt) / MW drug (free base) X target amount of free base Note: Occasionally two molecules of drug are included in the salt form, for example morphine sulfate. Check the chemical formula prior to making the standard. In the case of morphine sulfate, the molecular weight of the salt should be divided by 2 prior to using the formula above. Example: Diacetylmorphine HCl Monohydrate (heroin) C21H23NO5 HCl, H2O MW = 423.9 Diacetylmorphine (base) MW = 369.9 423.9 X 10 mg heroin base = 11.5 mg diacetylmorphine HCl monohydrate (heroin) 369.9 9. Calculation of drug as free base: Option 2 Alternatively, to calculate the percentage of free base in the salt molecule to produce a target amount, use the following equation: Example: Diacetylmorphine HCl Monohydrate (heroin) C21H23NO5 HCl, H2O MW = 423.9 Diacetylmorphine (base) MW = 369.9
% base in salt =
369.9 = 0.872 423.9
Amount of salt needed = 10.0 mg = 11.5 mg heroin 0.872
INFRARED SPECTROSCOPY Principle: Infrared spectroscopy provides unique identification of a substance when compared to a standard infrared spectrum. Molecules constantly vibrate, and chemical bonds within the compound stretch and bend with respect to each other. Molecules absorb infrared radiation that corresponds in energy to these vibrations. An infrared spectrometer measures the transmission of infrared radiation through a sample as the frequency of incident infrared radiation is varied over a known range. Instrument Calibration: Each month the instrument is used, the IR calibration should be checked using a polystyrene standard. The IR spectra of polystyrene should be compared with a known spectrum. Criteria for acceptable instrument operation are located in the IR Maintenance Log next to the IR. The analyst verifies instrument calibration by initialing and dating the spectrum which is filed for future reference. If the IR spectrum of polystyrene does not match the standard as noted, the analyst should tag the instrument as out of service and advise a supervisor immediately. Maintenance and repair is recorded in the IR Maintenance Logbook. Sample Preparation: Solid samples for identification by infrared spectroscopy are prepared and scanned using the Nujol mull technique. Hard samples may be reduced to a powder by grinding in a mortar while soft samples and powders may be used directly. About 5 mg of the powder is placed on a sodium chloride plate and a small drop of mineral oil is placed on another. The two plates are then ground together in a circular motion until the oil/powder mixture becomes smooth and transparent. More powder and/or oil can be added at anytime to adjust the transparency or concentration. Liquid samples can be used directly or in conjunction with mineral oil. If the scan is too concentrated, remove salt plates from mounting, slide plates apart, clean one plate with chloroform, rejoin plates, remount, and rescan. The salt plates are placed into an instrument mount, and screws are tightened to finger tightness. The mounting device is then placed in the instrument, and ready for scan. After scanning, the sodium chloride plates are cleaned by wiping on a paper towel wet with chloroform. Instrument Setup and Use: The instrument is turned on using the main power switch on the back, right hand side. Instrument is warmed up 5-10 minutes prior to use.
Infrared Spectroscopy Version 2.0
The instrument is set to the following parameters: Preset Scan Range = 4000 600 cm-1 - Chart Expansion: 0.5 - Scan Time/Slit Program: Wide: 3 - Adjust Chart to align with the Y-axis using Parameter Adjust: Forward or Reverse - Adjust Baseline/Offset between 70-90 using Parameter Adjust - Start Scan by pressing yellow Scan button Sample Identification: The IR spectrum of the unknown is compared to a spectrum of a known standard run in-house (see IR Standards Notebook). Identification may also be made using published spectral libraries such as the Aldrich Library of FTIR Spectra (Edition I, Vol 1 and 2); in this case, a note identifying the source of the reference spectra is placed in the case file.
Infrared Spectroscopy Version 2.0
LYSERGIC ACID DIETHYLAMIDE (LSD) ANALYSIS Principle of Assay: This analysis is designed to identify and quantitate lysergic acid diethylamide (LSD). LSD is identified by gas chromatography/mass spectrometry (GC/MS) and quantitated by gas chromatography (GC). The GC method separates LSD from its isomer lysergic acid methypropylamide (LAMPA). Sample Requirements: This method is suitable for analysis of LSD in a wide variety of matrices including blotter paper squares, candy, tablets, liquid solutions, and other types of evidence. It is imperative to understand the definition of abuse unit as described in the Texas Controlled Substances Act prior to analysis. Equipment: Gas chromatograph-mass spectrometer containing a methylsiloxane capillary column such as an HP-5MS, 30 m x 0.25 mm x 0.25 um film thickness and/or gas chromatograph-flame ionization detector with split/splitless injector containing a methylsiloxane capillary column such as an HP1, 15 m x 0.32 mm x 1.0 um film thickness (see GC/MS and/or GC Methods Notebook in Drug Laboratory). Screw top culture tubes, 25 mL, 20 mm x 125 mm, screw cap with Teflon liner Screw top culture tubes, 15 mL, 16mm x 125 mm, screw cap with Teflon liner Screw top conical tubes, 5 mL, screw cap with Teflon liner Autosampler vials, 32 mm x 11 mm, Teflon lined seals and glass inserts Pipettes, various Hamilton syringes, various Beaker, 50 mL Volumetric flasks, 5 mL, 25 mL Vortex mixer Sonicator Scissors Centrifuge Long range UV light source Reagents: Methanol, reagent grade or better Ethyl Alcohol, USP grade, 200 proof-Absolute Analytical Standards
Dallas County Institute of Forensic Sciences Controlled Substance Laboratory
LSD Analysis Version 2.0
Safety Precautions: The most common type of chemical or biological exposure in this type of laboratory is a splash to the skin or eye. Skin, mucous membranes, or eyes which have been splashed with commonly used chemicals or biologicals should be thoroughly washed for at least 15 minutes in cool tap water or eye wash station. Refer to appropriate MSDSs for additional chemical information. Report the incident immediately to a supervisor. Seek medical attention as necessary. Methanol and ethanol are flammable solvents. Avoid contact with skin, eyes, and mucous membranes. LSD is a hallucinogenic substance, which is active in small dosages. Wear gloves and use other precautionary procedures while handling the standard to limit accidental exposure during use. Avoid contact with skin, eyes, and mucous membranes. Internal Standard (I.S.) Solution Preparation: Internal Standard Amount of Drug (as HCl) Final Volume Solvent 0.5 mg/mL trazodone HCl 12.5 mg 25 mL ethanol Transfer 12.5 mg trazodone hydrochloride to a 25 ml volumetric flask and dilute to volume with ethanol. Mix by inverting. Sonicate as needed. Transfer to a 25 mL culture tube for storage. Calibration Standards: All solutions of stocks and standards are kept in the refrigerator. Allow the solutions to come to room temperature before using them. The concentration, amount, and volume of the calibration stock and calibration standards may vary proportionally depending on the availability of drug, and the expected concentration range of the drug. The following is provided as a guide. Calibration Stock Amount of Drug (as base) Final Volume Solvent 1.00 mg/mL LSD 10.0 mg 10 mL I.S. solution Transfer 10 mL of I.S. solution using a volumetric pipette to the dilutable bottle containing the drug. Mix by inverting bottle. Calibration Procedure: 1. 2. Choose a minimum of four appropriate calibration standards for the calibration curve based on the linear range required for the sample or drug to be analyzed. Using a Hamilton syringe, place the appropriate amount of calibration stock and I.S solution into a 5 mL conical screw cap tube. Vortex.
Calibration Standards 0.10 mg/mL LSD 0.25 mg/mL LSD 0.75 mg/mL LSD 1.00 mg/mL LSD
Amount of Calibration Stock 10 uL 25 uL 75 uL 100 uL
Volume I.S. Solution 90 uL 75 uL 25 uL 0
3. Transfer from the conical tube to an autosampler vial with a glass insert. 4. Inject into the GC using LSD.M method. 5. Determine if the data are acceptable using the instructions found in the Calibration Curve/Response Factor Log. a. An acceptable calibration curve has a correlation coefficient, r2 > 0.995. b. If the curve is acceptable, file it in the Calibration Curve/Response Factor Log located in the Drug Laboratory. If not, seek supervisory assistance as needed. Quality Control Standard: A quality control standard must be successfully run on each day this procedure is used. The concentration, amount, and volume may vary proportionally depending on the availability of drug. The following is provided as a guide: QC Stock Amount of Drug (as base) Final Volume Solvent 1.00 mg/mL LSD 10.0 mg 10 mL I.S. solution Transfer 10 mL of I.S. solution using a volumetric pipette to the dilutable bottle containing the drug. Mix by inverting bottle. Quality Control Procedure: 1. Using a Hamilton syringe, place the appropriate amount of QC stock and I.S solution into a 5 mL conical screw cap tube. Mix well. Amount of QC Stock 0.50 mL Volume of I.S. solution 1.50 mL
QC Standard 0.25 mg/mL LSD 2. 3.
Transfer from the conical tube to an autosampler vial with a glass insert. Inject into the GC using LSD.M method.
Evaluation Criteria: 1. 2. If the QC sample is within the set limits of +/- 10%, proceed with analysis. If the QC sample is not within the set limits of +/-10% of the target concentration, additional action must be taken to obtain an acceptable QC result such as re-running QC, re-prepping and re-runnning the QC, making new stock solution and re-analyzing, recalibrating (run curve), and/or consulting a supervisor.
LAMPA Standard: LAMPA Standard Amount of Drug (as base) Final Volume Solvent 1.0 mg/mL LAMPA 10 mg 10 mL I.S. solution Transfer 10 mL of I.S. solution using a volumetric pipette to the dilutable bottle containing the drug. Mix by inverting bottle. LSD/LAMPA Standard: LSD (0.5 mg/L) LAMPA (0.5mg/L) Combined Standard Amount of LSD QC Stock
0.5 mL Amount of LAMPA Standard 0.5 mL Transfer 0.5 mL LSD QC stock and 0.5 mL LAMPA standard to an autosampler vial. Mix well and inject using LAMPA.M method. Evaluation Criteria: 1. If the LSD/LAMPA peaks are resolved and identified on the chromatogram, include in case file. This LSD retention time is used for identification of sample. 2. Seek supervisory assistance if peaks are not resolved. Color Testing: 1. If sufficient sample is available, color test using the Van Urk reagent; for example, select 4 squares at random for testing. 2. In addition, long range UV light may indicate presence of drug on items such as candy and sugar cubes, etc. Analytical Procedure: Note: The final report should contain the weight of LSD quantitated in the exhibit(s) analyzed (when possible) and the number of abuse units as defined by the Texas Controlled Substances Act. Abuse unit is defined as follows: (A) except as provided by Paragraph (B): (i) a single unit on or in any adulterant, dilutant, or similar carrier medium, including, marked or perforated blotter paper, a tablet, gelatin wafer, sugar cube, or stamp, or other medium that contains any amount of a controlled substance listed in Penalty Group 1-A, if the unit is commonly used in abuse of that substance; or (ii) each quarter-inch square section of paper, if the adulterant, dilutant, or carrier medium is paper not marked or perforated into individual abuse units; or (B) if the controlled substance is in liquid form, 40 micrograms of the controlled substance
Dallas County Institute of Forensic Sciences Controlled Substance Laboratory 4 LSD Analysis Version 2.0
including any adulterant or dilutant. 1. Paper Squares, Sheets, Sugar Cubes, and Tablets: a. Use the information above to determine the number of abuse units in the exhibit to be analyzed. For paper sheets, the total area of exhibit must be determined. Record on the Drug Protocol Worksheet. b. Add 1-2 sugar cubes or tablets, 2 to 4 cut paper squares or quarter-inch square sections of paper into a 15mL screw cap culture tube. i. Record on the Drug Protocol Worksheet. c. Add 1-2 ml methanol, cap and sonicate for 20 minutes. d. Centrifuge for 5 minutes. e. Transfer methanol to a 5 mL conical test tube. f. Rinse culture tube with a few drops of methanol and add to the extract in conical test tube. g. Place conical test tube in warm water bath and evaporate to residue with air. h. Add at least 100-200 uL of trazodone internal standard solution and vortex to dissolve. i. Record the volume on the Drug Protocol Worksheet. i. Transfer to autosampler vial with glass insert. j. Inject the same vial on both GC and GC/MS using LSD.M method. k. Run LSD/LAMPA standard on the GC using LAMPA.M method; include chromatogram in case file. 2. Liquid Solutions: a. Record the total weight and volume on the Drug Protocol Worksheet. b. Determine the number of abuse units. i. Record on the Drug Protocol Worksheet. c. Place 40-50 uL into a 5 mL conical test tube. Record the volume on Drug Protocol Worksheet. d. Add 100-200 uL of trazodone internal standard solution and vortex to dissolve. i. Record the volume on Drug Protocol Worksheet. e. Transfer to autosampler vial with glass insert. f. Inject the same vial on both GC and GC/MS using LSD.M method. g. Run LSD/LAMPA standard on the GC using LAMPA.M method; include chromatogram in case file. 3. Sweetarts: a. Note: This procedure allows LSD identification but not quantitation. b. Determine the number of abuse units based on number of items (20 Sweetarts = 20 abuse units). i. Record on the Drug Protocol Worksheet. c. If long range UV light shows fluorescent spots, scrape fluorescent area from 2-3 items and place into 15 mL screw cap culture tube. d. If no fluorescent spots are visible, use 1-2 candies for extraction. e. Record materials tested on Drug Protocol Worksheet. f. Wash with three 5 ml portions of nBuCl: i. Add 5 mL solvent along with approximately 200 mg NaHCO3 to culture tube,
Dallas County Institute of Forensic Sciences Controlled Substance Laboratory LSD Analysis Version 2.0
g. h. i. j. k. l.
rotate 5 minutes, centrifuge and collect supernatant in 50 mL beaker. ii. Repeat process two more times. Take solvent to residue. Reconstitute with approximately 0.5 mL trazodone internal standard solution. Mix well by vortexing. Transfer to autosampler vial insert. Inject same vial on both the GC and GC/MS using LSD.M method. Run LSD/LAMPA standard on the GC using LAMPA.M method; include chromatogram in case file.
Instrument Parameters: See GC and GC/MS Methods Notebooks located in Drug Laboratory. Reporting Criteria: 1. The reporting range of this assay is +/-20% of the calibration curve. 2. If the concentration of LSD is below this range and an acceptable GC/MS spectra is obtained, reanalyze using more sample or report as present but insufficient for quantitation. 3. The final report should contain the weight of LSD quantitated in the exhibit(s) analyzed (if possible) and the number of abuse units as defined by the Texas Controlled Substances Act. Calculations: 1. Materials Analyzed by Weight or Volume (with exception of liquids) a. concentration of drug (mg/mL) x volume of internal standard solution used for extraction x 1000 ug/mg = ug LSD in the dosage unit(s) tested 2. Liquid Materials (only) a. concentration of drug (mg/mL) x volume of internal standard solution used for extraction x total weight of exhibit (mg) / total weight of sample analyzed (mg) x 1000 ug/mg = ug LSD 3. Dilutions a. In any situation in which a dilution is made, multiply the concentration of drug (mg/mL) by the dilution factor. A notation relating to how dilutions were made must be made in the case file.
Training Notes: LSD Analysis 1. A sonicator is necessary to bring materials into solution. Care should be used regarding the length of time materials are left in the sonicator since heat may decompose some drugs. Typically, a sonication time of 20 minutes has been found to be effective with regard to good recovery and minimal decomposition. 2. LAMPA and LSD are structural isomers. LAMPA is not scheduled. These substances are resolved by gas chromatography. 3. LSD is light sensitive and degrades with exposure to light. Protect standards from light by using amber bottles, covering bottles with aluminum foil, etc. 4. LSD may also be detected using long wave UV light.
MARIHUANA AND TETRAHYDROCANNABINOL ANALYSIS Principle of Assay: Marihuana is defined by the Texas Controlled Substances Act: Marihuana means the plant Cannabis sativa L., whether growing or not, the seeds of that plant, and every compound, manufacture, salt, derivative, mixture, or preparation of that plant or its seeds. The term does not include: (A) the resin extracted from a part of the plant or a compound, manufacture, salt derivative, mixture, or preparation of the resin; (B) the mature stalks of the plant or fiber produced from the stalks; (C) oil or cake made from the seeds of the plant; (D) a compound, manufacture, salt, derivative, mixture, or preparation of the mature stalks, fiber, oil, or cake; or (E) the sterilized seeds of the plant that are incapable of beginning germination. Non-marihuana material, such as hashish, which contains tetrahydrocannabinol is also scheduled. Marihuana contains more than 400 chemicals. Approximately 60 cannabinoids are present in the plant. (-) Delta 9- trans-tetrahydrocannabinol (levo isomer) is the principal psychoactive ingredient of the marijuana. This method describes a qualitative procedure for identification of marihuana from plant material and tetrahydrocannabinol from non-marihuana substances. Identification of marihuana is made using color test (Duquenois-Levine), macroscopic and microscopic examination, and GC/MS analysis for the presence of tetrahydrocannabinol (THC). Non-marihuana substances such as hashish - are identified using GC/MS to identify THC and positive Duquenois-Levine color test; microscopic analysis must exclude the material as marihuana. Equipment: Gas chromatograph-mass spectrometer (GC/MS) containing a methylsiloxane capillary column such as an HP-5MS, 30 m x 0.25 mm x 0.25 um film thickness with split/splitless injector (see GC/MS Notebook in Drug Laboratory). Stereomicroscope Analytical and top-loading balance Disposable culture test tubes, 12 mm x 100 mm Autosampler vials, 32 mm x 11 mm, Teflon lined seals Vortex mixer Disposable test tubes, 16 x 100 mm Beaker, Parafilm, and paper towels for seed germination
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory Marihuana Analysis Version 2.0
Reagents: Methanol, reagent grade or better Ethanol, reagent grade or better Duquenois-Levine reagent (see Color Testing Procedure) Hydrochloric acid, reagent grade or better Chloroform, reagent grade or better Safety Precautions: The most common type of chemical or biological exposure in this type of laboratory is a splash to the skin or eye. Skin, mucous membranes, or eyes which have been splashed with commonly used chemicals or biologicals should be thoroughly washed for at least 15 minutes in cool tap water or eye wash station. Refer to appropriate MSDSs for additional chemical information. Report the incident immediately to a supervisor. Seek medical attention as necessary. Methanol is a flammable solvent. Avoid contact with skin, eyes, and mucous membranes. Concentrated hydrochloric acid is a corrosive. Avoid contact with skin, eyes, and mucous membranes. Vapors are also corrosive and will corrode metal. Use concentrated acid in a hood. Chloroform is a chlorinated solvent and is considered a human carcinogen. Use in a hood or well ventilated area. Avoid contact with the skin, eyes, and mucous membranes. Overgrowth of mold and presence of mold spores may occasionally be observed in marihuana cases. If these cases must be analyzed, processing must be done in a hood. Seek supervisory assistance as needed. Sample Requirements: Plant materials, oils, solids, residues, and other types of drug evidence are suitable for analysis using this procedure. Plant material should be dried prior to storage to prevent decomposition. Sample Preparation: Obtain the total weight of the suspected drug material minus the packaging. Remove stems or stalks greater than inch in diameter and note on the Marihuana Identification Protocol sheet. Color Testing: See section on Color Testing for preparation of Duquenois-Levine reagent. Quality Control Procedure: This is a qualitative procedure. Results of testing are compared to known characteristics of marihuana or tetrahydrocannabinol verified in this Laboratory using known marihuana or
tetrahydrocannabinol. Analytical Procedure: 1) Macroscopic Observation a) Marihuana is typically submitted to the Laboratory as crumbled, dried, green-brown, leafy material. b) Marihuana leaves i) compound each being made from 3-11 leaflets most commonly 7 leaflets and almost always an odd number of leaflets; leaflets are about 6 times as long as broad ii) palmate hand-like structure radiating out from a common point at the end of the stem. iii) serrated along the edge pointing toward the sharp tip of each leaflet iv) darker green on the upper surface and lighter green on the underside c) Seeds i) covered by hard shell ii) ellipsoid or oval in shape and slightly compressed or flattened at one end iii) generally grey-brown with mottled appearance iv) about 2.5 to 5 mm long and 2 to 3.5 mm in diameter v) germinates within 3-7 days under proper conditions d) Stem/Stalk i) angular ii) sometimes hollow iii) covered with curved cystolythic hairs which appear pressed against the stem when viewed microscopically 2) Microscopic Analysis a) A stereomicroscope is used to observe plant structures which are consistent with marihuana. The sample is viewed at varying magnifications (approximately 10-40X). i) Cystolithic hairs - The upper surface of the marihuana leaf contains unicellular, sharply pointed, and curved hairs with enlarged bases containing cystoliths of calcium carbonate. These hairs are shaped like a bear claw and called cystolithic hairs. Cystolithic hairs are found on all parts of the plant including the roots. They are more abundant on the flowering plant and on the upper leaves. Cystolithic hairs are not found on seeds or bracts. ii) Simple hairs The surface of the underside of the leaf contains conical hairs which do not taper. They are usually longer, more slender, and more numerous than cystolithic hairs. iii) Glandular hairs These hairs may appear on the upper and lower leaf surface when the plant is about to flower. Hairs are multi-cellular, shiny, brown or yellowish, bulbous at the tip, and are covered with a sticky resin. (a) Although other plants may have cystolithic hairs or simple hairs, marihuana is the only plant which has cystolithic hairs on one side of the leaf and simple hairs on the other side. Glandular hairs are not always present in marihuana. 3) Duquenois-Levine Color Test: a) The Duquenois- Levine color test yields a characteristic color when applied to marihuana
b) c) d) e) f) g) h) i)
plant material, hashish, other marihuana preparations, and tetrahydrocannabinol. Add approximately 25 mg of suspected material to a disposable test tube. Add approximately 2 mL Duquenois-Levine reagent. Mix and let settle. Add an approximately equal amount of concentrated hydrochloric acid. Mix. i) A deep blue-purple color will develop with marihuana or tetrahydrocannabinol. Add enough chloroform to form a visible second layer of liquid. Mix and allow to stand until the organic and aqueous layers separate. A positive test results when the purple color extracts into the bottom (organic) layer. i) Note: This test may also be performed on the dried petroleum ether extract of plant material.
4) GC/MS Analysis: a) Place approximately 0.1 g plant material, 1 drop of oil, or aliquot of specimen into a disposable test tube. i) The analyst will vary the amount of material as needed to obtain an acceptable mass spectrum. b) Add enough methanol to cover sample or dilute to an appropriate concentration for analysis. c) Mix and allow to separate. d) Place an aliquot of the methanol into an autosampler vial and inject onto the GC/MS for the identification of tetrahydrocannabinol. 5) Seed Germination a) Seed germination is performed upon request or when the analyst judges it to be appropriate. i) If a submitted exhibit is primarily seeds and not plant material, the analyst should discuss the case with a supervisor. b) Label a beaker with the case and exhibit number. c) Take approximately 5-10 seeds randomly from the marihuana exhibit and place between two sheets of moist paper towel. Fold the paper towel into a packet with the seeds inside and place into a beaker. Cover the beaker with Parafilm. d) Place the beaker in a dark evidence locker at room temperature for about 3-7 days. e) Inspect the seeds to determine if germination has occurred. f) If germination occurs i) Photograph the seeds and place the photo in the case file. (1) Label the photo with the case number, exhibit number, date, and your initials; include a ruler in the photo if applicable. (2) Include the weight of seeds in the weight of marihuana reported. g) If no germination occurs i) Take approximately one ounce of marihuana from the exhibit. ii) Weigh the sample. iii) Separate the seeds. iv) Weigh the seeds. v) Calculate the fraction of seeds in the marihuana sample.
vi) Multiply the gross weight of marihuana by this percent to obtain the total amount of non-germinable seeds. vii) Subtract this from the gross weight of marihuana to obtain the net weight of marihuana without germinable seeds. viii) Calculations follow: (1) (weight of seeds from plant material sample)/(weight of plant material sample) = fraction of seeds in plant material (2) fraction of seeds in plant material x gross weight of marihuana = gross weight of non-germinable seeds (3) gross weight of marihuana gross weight of non-germinable seeds = net weight of marihuana without non-germinable seeds 6) Non-Marihuana Substances Containing THC a) Examples of non-marihuana substances containing THC are b) Hashish/hash - a THC rich resinous material isolated from marihuana. It varies in color from light tan, green, brown, or black and in texture from soft to very hard. Microscopically, hashish contains free or unattached cystolithic hairs, simple hairs, and glandular hairs; it may also contain small fragments of torn leaf tissue. c) Hash oil - a viscous liquid made by solvent extraction of marihuana to concentrate THC and other cannabinoids. It is usually a viscous liquid which may range from amber to dark brown in color. Hash oil is typically free of plant fragments. d) Microscopic Analysis - These materials will not show typical microscopic morphology for marihuana. Although, hashish will usually contain characteristic hairs from marihuana, there will be very little intact leaf material present. i) The analyst must use microscopy to distinguish between powdered marihuana and hashish. Powdered marihuana will usually contain intact leaf fragments containing cystolithic and simple hairs consistent with characteristic marihuana morphology. Powdered marihuana is reported as marihuana; hashish is reported as containing tetrahydrocannabinol. It is recommended that these cases be reviewed with a supervisor. e) Duquenois-Levine and GC/MS The Duquenois-Levine will be positive and GC/MS will be positive for THC and typically either cannabinol or cannabidiol. f) Reporting - Non-marihuana substances which contain THC are reported as containing tetrahydrocannabinol. 7) Pipe Residue a) Rinse the pipe residue with methanol. b) Place methanol in a GC/MS vial and analyze for THC. c) Return the contents of the GC/MS vial to the residue tube and dry down. d) Perform the Duquenois-Levine test on the residue. e) If the Duquenois-Levine is positive and THC is present, the case is reported: the charred residue in the pipe contained tetrahydrocannabinol. Instrument Parameters: See GC/MS Methods Notebook located in the Drug Laboratory.
Reporting Criteria: 1. Marihuana is identified if both the microscopic and Duquenois-Levine tests are positive for marihuana. Tetrahydrocannabinol in marihuana is identified by GC/MS; at least one GC/MS will be run on each marihuana case. a. If the material appears grossly like plant material, the microscopic analysis is inconclusive, THC is present by GC/MS, and the Duquenois-Levine is positive for marihuana, perform the microscopic analysis on another aliquot of material. If it is still inconclusive, review the case with a supervisor. 2. Tetrahydrocannabinol is identified if marihuana plant material is not present, the Duquenois-Levine is positive, and tetrahydrocannabinol is identified by GC/MS. 3. Exhibits consisting primarily of seeds not plant material should be reviewed with a supervisor prior to analysis.
Marihuana Analysis Version 2.0
Training Notes: Marihuana Analysis 1. Fresh marihuana plants that have been stored in non-permeable containers decompose. A positive test for marihuana may not be possible under these conditions. Photograph case where possible. 2. Penalty for possession or delivery of marihuana in Texas is covered by marihuanaspecific sections of the Texas Controlled Substances Act. Tetrahydrocannabinol is included in Penalty Group 2. 3. Dronabinol (trade name Marinol) is a commercial pharmaceutical containing THC in sesame oil in a round, soft gelatin capsule. Refer to the steroid method for analysis. Review case with a supervisor. Dronabinol is included in Penalty Group 2. 4. Hashish, hash oil, and other non-marihuana materials containing THC are not specifically covered by name in the Texas Controlled Substances Act. They are controlled by virtue of the fact that they contain THC. 5. Overgrowth of mold and presence of mold spores may occasionally be observed in marihuana cases. If these cases must be analyzed, processing must be done in a hood. Seek supervisory assistance as needed. Photograph case as applicable. 6. Plant material is weighed in the condition it is received; it is typically not dried before processing. If the material is fresh or likely to decompose further in the analysts judgment, it is recommended that a photo be taken and included in the case file. 7. When a case contains numerous exhibits, the analyst may choose to run more than one GC/MS, for example one GC/MS for each 10 items. 8. In some cases (immature and old plant material), it may be necessary to extract the plant material with petroleum ether, evaporate to dryness with heat, and color test the residue.
Marihuana Analysis Version 2.0
MISCELLANEOUS MATERIALS Principle: In general, positive identification of materials is done by gas chromatography/mass spectrometry (GC/MS) or infrared spectroscopy (IR). This section concerns procedures identifying controlled substances in miscellaneous materials such as GHB, mushrooms, steroids, and volatiles. Equipment: Gas chromatograph-mass spectrometer containing a methylsiloxane capillary column such as an HP-5MS, 30 m x 0.25 mm x 0.25 um film thickness and/or gas chromatograph-flame ionization detector with split/splitless injector containing a methylsiloxane capillary column such as an HP1, 15 m x 0.32 mm x 1.0 um film thickness (see GC/MS and/or GC Methods Notebook in Drug Laboratory). IR Spectrophotometer with accessories Screw top culture tubes, 15 ml, 16 mm x 125 mm, screw cap with Teflon liner Test tubes Vortex mixer Sonicator Centrifuge Pipettes, various Evaporating dishes (watch glass) Spot test plates pH paper Hot plate Reagents: Methanol, reagent grade or better Hexane, reagent grade or better Acetonitrile, reagent grade or better Safety Precautions: The most common chemical exposure in this type of laboratory is a chemical splash to the skin or eye. Skin, mucous membranes, or eyes which have been splashed with commonly used chemicals or biologicals should be thoroughly washed for at least 15 minutes in cool tap water or eye wash station. Refer to MSDSs for additional chemical information. Report the incident immediately to a supervisor. Seek medical attention as necessary. Methanol and hexane are flammable solvents. Use in a hood or in a well ventilated area. Acetonitrile is a suspected teratogen as well as a flammable solvent. Use in a hood or in a well ventilated area. Avoid contact with skin.
Dallas County Institute of Foresic Sciences Controlled Substances Laboratory Miscellaneous Materials Version 2.0
Color Testing: See the section on Color Testing. Spot tests will usually be run on each item of nonpharmaceutical evidence within one case up to the applicable statutory weight limit and will be performed on pharmaceutical evidence as applicable. GHB Analytical Procedure: 1) Liquid Samples: a) Obtain the total weight and the total volume of the material in the exhibit to be analyzed and record on the Drug Protocol Worksheet. b) Color test with ferric chloride. i) An orange color is an indication that GHB is present. c) GC/MS Identification i) Transfer approximately 10-20 drops of liquid to be analyzed to approximately 1.0 mL methanol in a test tube. ii) Transfer an aliquot to an autosampler vial and inject on the GC/MS using BLANK.M method. iii) GHB breaks down to GBL at high temperatures on the GC. (1) The presence of GBL is consistent with the presence of GHB; however, confirmation by IR is required to conclusively determine whether GHB or GBL is present. d) IR Identification i) Obtain and record tare weight of a watch glass on Drug Protocol Worksheet. ii) Add several drops of sample to a watch glass so that the spot is approximately the diameter of a quarter, obtain weight of watch glass plus sample and record on the Drug Protocol Worksheet. iii) Place watch glass on hot plate using low heat. Evaporate sample to dryness. iv) Allow watch glass to cool for several minutes. v) Obtain weight of watch glass plus dry sample and record on the Drug Protocol Worksheet. vi) Analyze dry sample by IR; refer to IR procedure. vii) Calculate the total weight of GHB. 2) Liquid Samples Containing Sugars: a) Obtain the total weight and the total volume of the material in the exhibit to be analyzed and record on the Drug Protocol Worksheet. b) Color test with ferric chloride. i) An orange color is an indication that GHB is present. c) GC/MS Identification i) Transfer approximately 10-20 drops of liquid to be analyzed to 1.0 mL methanol in a test tube. ii) Transfer an aliquot to an autosampler vial and inject on the GC/MS using BLANK.M method.
Dallas County Institute of Foresic Sciences Controlled Substances Laboratory
Miscellaneous Materials Version 2.0
(1) The presence of GBL is consistent with the presence of GHB; however, confirmation by IR is required to conclusively determine whether GHB or GBL is present. d) IR Identification i) Obtain and record tare weight of a watch glass on Drug Protocol Worksheet. ii) Add several drops of sample to a watch glass so that the spot is approximately the diameter of a quarter, obtain weight of watch glass plus sample and record on the Drug Protocol Worksheet. iii) Place watch glass on hot plate using low heat. Evaporate sample to dryness. iv) If the sample does not evaporate, resample. (1) Place 3-5 mL of sample in methanol and filter to remove insoluble components. (2) Transfer liquid to test tube and evaporate to a volume of approximately 1 mL. (3) Add 5-10 mL acetone to precipitate the GHB salt, decant the solvent. (4) Wash the precipitate with approximately 1 mL acetone, evaporate, and proceed with IR. e) Reporting i) GHB liquids containing sugar can not be quantitated. Report presence of GHB and total weight of liquid in exhibit. High and Low pH Liquids: Clear aqueous liquids which are extremely acidic or basic to pH paper do not usually contain controlled substances and typically are not analyzed. Volatile Liquids: Volatile liquids are commonly called inhalants; these materials are generally organic solvents and nitrites. They can be identified by gas chromatography/mass spectrometry (GC/MS) and/or infrared spectroscopy (IR). Identification is made by comparison to standard materials. Organic nitrites decompose on the injection block of the GC/MS to the corresponding alcohol. The GC/MS can be used to determine the alcohol part of unknown alkylnitrites. Specific identification of the identity of organic nitrites is made by IR; however, identification may not be possible for a mixture of organic nitrites. Since volatile compounds evaporate rapidly, a smear test can be used as a presumptive test. Place a drop of liquid on a hard surface in a hood and smear with gloved finger. If material evaporates quickly, a volatile may be present. To identify the volatile content of spray paints and other aerosols, spray a small amount into a screw cap vial containing 3 mL of methanol. Shake well and allow settling. Place an aliquot of the clear supernatant in an autosampler vial and inject on the GC/MS using VCHEM.M method. Inhalant paraphernalia such as empty soda cans, empty bottles, rags, and plastic bags can often be successfully analyzed by rinsing with methanol and injecting an aliquot on GC/MS using VCHEM.M method.
Steroids Analytical Procedure: Steroids are almost always encountered in dosage forms and in general can be classified as tablets/capsules, aqueous injectable solutions, and vegetable oil injectable solutions. 1) Steroids in Oil: a) Obtain the total weight of the material in the exhibit to be analyzed and record on the Drug Protocol Worksheet. b) Place approximately 6 drops of the oil into a 1:1 mixture of hexane:acetonitrile. c) Mix well by vortexing. d) Transfer the acetonitrile layer (bottom) to an autosampler vial and inject on the GC/MS using STEROID.M method. 2) Tablets, Capsules and Other Pharmaceutical Preparations: a) Obtain the total weight of the material in the exhibit to be analyzed and record on the Drug Protocol Worksheet. b) Select one tablet or capsule from a population as a representative sample for analysis. Grind to a powder if necessary. c) GC/MS Identification i) Transfer approximately 10 mg of the sample to 1.0 mL methanol in a test tube. ii) Vortex and centrifuge as necessary. iii) Transfer an aliquot to an autosampler vial and inject on the GC/MS using STEROID.M method. 2) Liquids: a) Obtain the total weight of the liquid in the exhibit to be analyzed and record on the Drug Protocol Worksheet. b) GC/MS Identification i) Place approximately 1 part sample to 4 parts methanol in a 15 mL culture tube tube. Mix well before sampling. ii) Transfer an aliquot to an autosampler vial. Analyze by GC/MS using STEROID.M method. Mushrooms/Cactus Bulbs Analytical Procedure: Mushrooms and cactus buttons are the most common plant materials other than marihuana submitted for identification. These can be identified by gas chromatography/mass spectrometry (GC/MS) and are not quantitated. 1) Mushrooms: a) Obtain the total weight of the material in the exhibit to be analyzed and record on the Drug Protocol Worksheet. b) Locate the part of the mushroom that has a purple/blue tint (usually on the stem) and proceed with color testing using the Van Urk reagent. Record results on Drug Protocol Worksheet. c) Chop up approximately 500 mg of material (purple/blue portion). d) Place in a screw cap culture tube.
Add enough methanol to cover the material. Sonicate for 20 minutes. Centrifuge as necessary. Decant the methanol into another tube. Concentrate the methanol down to approximately 1 mL. Transfer the methanol extract to an autosampler vial and inject on the GC/MS using LSD.M method. 2) Cactus bulbs (buttons): a) Obtain the total weight of the material in the exhibit to be analyzed and record on the Drug Protocol Worksheet. b) Chop or grind up approximately 500 mg of material. c) Place in a screw cap culture tube. d) Add enough methanol to cover the material. e) Sonicate for 20 minutes. f) Centrifuge as necessary. g) Decant the methanol into another tube. h) Concentrate the methanol down to approximately 1 mL. i) Transfer the methanol extract to an autosampler vial and inject on the GC/MS using LSD.M method. Instrument Parameters: See GC and GC/MS Methods Notebooks located in Drug Laboratory. Reporting Criteria: 1) Report the identity of the drug when an acceptable GC/MS and/or IR is obtained . 2) Psilocin and psilocybin cannot be distinguished by GC/MS, therefore report presence of both, i.e. psilocin/psilocybin.
e) f) g) h) i) j)
Training Notes: Miscellaneous Materials 1. A sonicator may be used to bring materials into solution. Care should be used regarding the length of time materials are left in the sonicator since heat may decompose some drugs. Typically, a sonication time of 5 minutes or less is used. 2. Seek supervisory assistance and/or refer to the Microgram Bulletin (July-December 2003) for mushroom/chocolate concoctions. 3. Mushrooms should be analyzed immediately. 4. For aqueous steroid injectables, take to residue with heat if necessary, and extract the solid residue with methanol and analyze by GC/MS. 5. The Van Urk produces subtle color changes and is often inconclusive. 6. GHB converts to GBL on the injection block of GC/MS.
MORPHINE SYRUP ANALYSIS Principle of Assay: This method describes the analysis of morphine in syrup formulations. This drug is amphoteric and has both acidic and basic functional groups. Extraction of this substance from an aqueous matrix into an organic phase requires that the pH must be above the pKa for the acidic OH group and below the pKa for the amine function. The weakly basic drug is dissolved in water and the pH of the solution is made weakly basic (approximately pH 8.5) using sodium bicarbonate/sodium carbonate mix. In the weakly basic aqueous solution, the weakly basic drug is converted from its ionic salt form to its nonionic free base form. In this form, the drug is quantitatively extracted into ethyl acetate/isobutanol (9:1) containing an internal standard. Quantitative analysis is performed on the extracted drug using gas chromatography (GC). Morphine is identified using gas chromatography/mass spectrometry (GC/MS). Sample Requirements: Liquids are suitable for analysis using this procedure. Equipment: Gas chromatograph-mass spectrometer containing a methylsiloxane capillary column such as HP-5MS, 30 m x 0.25 mm x 0.25 um film thickness and/or gas chromatograph-flame ionization detector with split/splitless injector containing a methylsiloxane capillary column such as HP-1, 15 m x 0.32 mm x 1.0 um film thickness (see GC/MS and/or GC Methods Notebooks in Drug Laboratory). Screw top culture tubes, 25 mL, 20 mm x 125 mm, screw cap with Teflon liner Screw top culture tubes, 15 mL, 16 mm x 125 mm, screw cap with Teflon liner Volumetric flasks, 10 mL, 25 mL, 100 mL, 2 liter Pipettes, various Graduated cylinders, various Autosampler vials, 32 mm x 11 mm, Teflon lined seals and glass inserts Vortex mixer Rotator Centrifuge Analytical balance pH paper Reagents: Sodium bicarbonate, reagent grade or better Sodium carbonate, reagent grade or better
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory Morphine Syrup Analysis Version 2.0
Ethyl acetate, reagent grade or better Isobutanol, reagent grade or better Ethyl Alcohol, USP grade, 200 proof - Absolute Analytical standards Carbonate mix Reagent Amount of Chemical Carbonate Mix (8:3) 80 g sodium bicarbonate + 30 g sodium carbonate Pulverize with a mortar and pestle. Store at room temperature. Ethyl acetate/isobutanol (9:1) Reagent Volume of Solvent Total Volume Ethyl acetate/ isobutanol (9:1) 90 mL ethyl acetate + 10 mL isobutanol 100 mL Pour 90 mL of ethyl acetate and 10 mL isobutanol into a 100 mL flask. Mix by inverting the flask. Safety Precautions: The most common type of chemical or biological exposure in this type of laboratory is a splash to the skin or eye. Skin, mucous membranes, or eyes which have been splashed with commonly used chemicals or biologicals should be thoroughly washed for at least 15 minutes in cool tap water or eye wash station. Refer to appropriate MSDSs for additional chemical information. Report the incident immediately to a supervisor. Seek medical attention as necessary. Ethyl acetate and isobutanol are volatile solvents. Avoid contact with skin; do not breathe vapors. Internal Standard (I.S.) Solution Preparation: Internal Standard Amount of Drug (as base) Final Volume Solvent 0.5 mg/mL codeine 50 mg 100 mL (9:1) ethyl acetate:isobutanol Transfer 50 mg codeine calculated as the free base to a 100 mL volumetric flask, and bring to volume with solvent. Calibration Standards: All solutions of stocks and standards are kept in the refrigerator. Allow the solutions to come to room temperature before using them. The concentration, amount, and volume of the calibration stock and calibration standards may vary proportionally depending on the availability of drug and the expected concentration range of the drug. The following is provided as a guide:
Calibration Stock Amount of Drug (as base) Final Volume Solvent 1.0 mg/mL morphine 10.0 mg 10 mL deionized water Transfer 10.0 mg morphine calculated as the free base to a 10 mL volumetric flask and bring to volume with solvent. Mix by inverting the flask. Sonicate as needed. Transfer to a 15 mL culture tube for storage. Calibration Procedure: 1. Choose a minimum of three appropriate calibration standards for the calibration curve based on the linear range required for the sample or drug to be analyzed. 2. Place the appropriate amount of calibration stock into a 15 mL labeled, screw cap culture tube using a volumetric pipette. Add deionized (DI) water to make a total volume of 5.0 mL. Add approximately 200 mg carbonate mix and 5.0 mL I.S. solution. Calibration Standards 0.20 mg/mL 0.50 mg/mL 1.00 mg/mL 3. 4. 5. 6. 7. Amount of Calibration Stock 1.0 mL 2.5 mL 5.0 mL Amount of DI water 4.0 mL 2.5 mL 0 Volume I.S. solution 5 mL 5 mL 5 mL
Cap the tubes and place on a rotator for 5 - 10 minutes. Centrifuge for 5 - 10 minutes on medium setting. Transfer an aliquot of the organic layer to an autosampler vial. Inject the extracted calibration standards into the GC using SYRMORPH.M method. Determine if the data are acceptable using the instructions found in the Calibration Curve/Response Factor Log. An acceptable calibration curve has a correlation coefficient, r2 > 0.995. If the curve is acceptable, file it in the Calibration Curve/Response Factor Log located in the Drug Laboratory. If not, seek supervisory assistance as needed.
Quality Control Standard: Ideally the quality control stock is made from a different lot number or manufacturer than the calibration stock. At a minimum, it should be made from a different stock solution. A quality control standard must be successfully run on each day this procedure is used. The concentration, amount, and volume may vary proportionally depending on the availability of drug. The following is provided as a guide: QC Stock 1.0 mg/mL morphine Amount of drug (as base) 10.0 mg Final Volume 10 mL Solvent deionized water
Morphine Syrup Analysis Version 2.0
Transfer 10.0 mg drug calculated as the free base to a 10 mL volumetric flask and bring to volume with solvent. Mix by inverting the flask. Sonicate as needed. Transfer to a 15 mL culture tube for storage. Quality Control Procedure: 1. Place the appropriate amount of QC stock into a 15 mL labeled, screw cap culture tube using a volumetric pipette. Add deionized (DI) water to make a total volume of 5.0 mL. Add approximately 200 mg carbonate mix and 5.0 mL I.S. solution. QC Standard 0.5 mg/mL morphine 2. 3. 4. 5. Amount of QC Stock 2.5 mL Amount DI water 2.5 mL Volume I.S. solution 5 mL
Cap the tubes and place on a rotator for 5 - 10 minutes. Centrifuge for 5 - 10 minutes on medium setting. Transfer an aliquot of the organic layer to an autosampler vial. Inject the extracted QC standard into the GC using SYRMORPH.M method.
Evaluation Criteria: 1. If the QC sample is within the set limits of +/- 5% of the target concentration, proceed with analysis. 2. If the QC sample is not within the set limits of +/- 5% of the target concentration, additional action must be taken to obtain an acceptable QC result such as re-running QC, re-extracting and re-running QC, making new stock solution and new QC with reextraction and reanalysis, recalibrating (run curve), and/or consulting a supervisor. Color Testing: See the section on Color Testing. Spot tests will usually be run on each item of nonpharmaceutical evidence within one case up to the applicable statutory weight limit. Analytical Procedure: Liquid samples may be analyzed by weight or by volume. If the sample is to be analyzed by volume, a known volume of the liquid must be weighed to determine density. This information is recorded on the Drug Protocol Worksheet 1. Obtain the total volume and weight of liquid in the exhibit and record on the Drug Protocol Worksheet. 2. GC/MS Identification a. Place approximately 1 part sample to 4 parts methanol, in a 15 mL screw cap culture tube.
i. The analyst will vary the amount of material and methanol as needed to obtain an acceptable mass spectrum. b. Transfer an aliquot to an autosampler vial. c. Analyze by GC/MS using BLANK.M method. 3. GC Quantitation a. Transfer a known volume and weight of the sample to a 15 mL labeled screw top culture tube and record on the Drug Protocol Worksheet. b. Add deionized water to make a total volume of 5.0 mL. c. Add approximately 200 mg carbonate mix and 5.0 mL internal standard solution to the culture tube. d. Cap the tube and place on rotator for 5 - 10 minutes. e. Centrifuge for 5 -10 minutes on medium setting. f. Transfer an aliquot of the organic layer to an autosampler vial. g. Inject the extracted sample into GC using SYRMORPH.M method and calculate the results. Instrument Parameters: See GC and GC/MS Methods Notebooks in Drug Laboratory. Reporting Criteria: 1. Report both the quantity and the identity of the drug when the concentration of the drug is within +/- 20% of the calibration curve on the GC and the GC/MS is complete. It may be necessary to dilute extracted samples to bring them within curve range. 2. Report the amount of the drug as insufficient for quantitation when the drug concentration is below 20% of the calibration curve on the GC and the GC/MS is complete. 3. Since penalty group is determined by the concentration of this drug, report the amount of drug in mg/100 mL. Calculations: 1. Dilutions a. In any situation in which a dilution is made, multiply the concentration of drug (mg/mL) by the dilution factor. A notation relating to how dilutions were made must be made in the case file. 2. Materials Analyzed by Volume a. concentration of drug (mg/mL) x volume of internal standard solution used for extraction (mL) x total exhibit volume (mL)/extracted sample volume (mL) = mg of drug in the exhibit as the free base 3. Materials Analyzed by weight: a. concentration of drug (mg/mL) x volume of internal standard solution used for
extraction (mL) x total exhibit weight (mg)/extracted sample weight (mg) = mg of drug in the exhibit as the free base 4. Amount drug (mg/100 mL): a. mg / 100 mL = mg (drug) X 100 Total Volume of exhibit
Training Notes: Morphine Syrup Analysis 1. If GC/MS results are negative when run on BLANK.M, run on alternative GC/MS methods such as LOWDOSE.M, LSD.M, etc. until satisfied that no controlled substance is present. 2. Results should be reported in mg/100 mL. Penalty group is based on number of mg/100 mL. 3. Refer to Direct Dilution method for residues and morphine in other matrices. 4. The color tests may be inconclusive based on the concentration of the sample or the color of the liquid. 5. Calculation of drug as free base: Option 1 To calculate the amount of drug as a salt required to produce a target amount of free base, use the following equation: MW drug (salt) / MW drug (free base) X target amount of free base Note: Occasionally two molecules of drug are included in the salt form, for example morphine sulfate. Check the chemical formula prior to making the standard. In the case of morphine sulfate, the molecular weight of the salt should be divided by 2 prior to using the formula above. Example: Morphine sulfate (C17H19NO3)2 H2SO4 . 5H2O MW = 758.8 Morphine (base) MW = 303.4 758.8 / 2 X 10 mg morphine base = 12.5 mg morphine sulfate 303.4 6. Calculation of drug as free base: Option 2 Alternatively, to calculate the percentage of free base in the salt molecule to produce a target amount, use the following equation: Example: Morphine sulfate (C17H19NO3)2 H2SO4 . 5H2O MW = 758.8 Morphine (base) MW = 303.4 303.4 X 2 % base in salt = 758.8
=
0.799
Amount of salt needed = 10.0 mg = 12.5 mg morphine sulfate 0.799
OFF-SITE SAMPLING OF EVIDENCE General Information: This procedure outlines the process of off-site sampling of suspected controlled substance cases which is usually performed at a submitting agency property room and submission of core samples to the Controlled Substances Laboratory for examination. This process is employed when it is not feasible to submit evidence directly to the Laboratory for analysis. The most common type of controlled substance sampled off-site is a large marihuana seizure; therefore, this procedure primarily focuses on marihuana cases, but it may be used for other suspected controlled substance cases. Notification: To initiate a request for off-site sampling, a submitting agency submits a Notice of Marihuana Core Sample sheet to the Drug Analysis Laboratory Evidence Registrar typically by fax or in person. The Evidence Registrar notifies an IFS chemist of the request, issues a unique forensic laboratory case number to the case, prints IFS evidence labels, and transfers the Notice of Marihuana Core Sample sheet to the assigned chemist. The IFS Chemist contacts the submitting agency to make an appointment for off-site sampling. Fresh plant material should be sampled as soon as possible, ideally on the next business day. Transportation: Employees of the Southwestern Institute of Forensic Sciences do not transport suspected drug evidence. Core samples are left at the property or with law enforcement officers for submission at a later time. Mileage reimbursement for business travel to and from the off-site location is handled per standard County procedures. Equipment and Supplies: The chemist transports the following equipment and supplies to the off-site location in or with the core sample carrying case: 1. Notice of Marihuana Core Sample sheet (obtained from Supervisor or Evidence Registrar) 2. IFS evidence labels containing a unique forensic laboratory number to be assigned to the case (obtained from the Evidence Registrar) 3. Evidence Summary sheet for additional items 4. Certified weights of appropriate mass, typically one 1 kg and one 5 lb weight 5. Digital camera, charged with space on disc
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory 1 Off-Site Sampling Version 2.0
6. Ziplock bags for sampling 7. Evidence bags 8. Evidence container labels to identify evidence bags and document chain of custody 9. Packaging tape 10. Evidence tape 11. Duct tape 12. Manila envelopes 13. Marihuana protocol sheets 14. Box cutters or knives 15. Pens and sharpies 16. Calculator 17. Various office supplies such as scissors 18. Kimwipes 19. Gloves 20. Screwdriver 21. Roll of shrink wrap 22. Goggles, plastic aprons, and plastic sleeves Verification of Balance Operation: The Chemist must first verify proper operation of the balance at the off-site location using an IFS certified weight and the Laboratorys standard balance verification procedure. The Analyst makes note of the certified weight used, the actual weight measured by the balance, and the difference found between actual and expected on the Notice of Marihuana Core Sample sheet. If the difference between the actual and expected weight is outside the accepted range of +/- 3 units, the Chemist will inform property room staff of the problem, cease sampling, and request notification when the balance has been properly calibrated. Sampling: IFS staff will follow Laboratory standard operating procedures when weighing and processing the suspected controlled substance. Sampling Procedure: 1. Establish Chain of Custody for primary containers with the submitting agency using the Notice of Marihuana Core Sample sheet. 2. Describe and count the number of primary containers and record them on the Notice of Marihuana Core Sample sheet. 3. Place the unique forensic laboratory number on each primary container. 4. Verify that each container is in good condition and properly sealed. a. If containers are not properly sealed or are in poor condition, report it to the property room staff and document this on the Notice of Marihuana Core Sample sheet. i. Determine whether sampling can continue.
5. Weigh each primary container and record the gross weight. 6. Begin inventory and sampling. a. Determine the number of items it will take to be within a state or federal weight range. i. State and Federal regulations on sample weight will gauge the extent of sampling. b. Obtain photographic documentation of each exhibit. c. Inventory and obtain a core sample of all items needed to establish a weight within the highest punishment level. i. When determining which items to sample, always choose the bulkiest and most abundant exhibits first. ii. Within each primary container, sort all items by content and packaging. iii. Record gross weights of like items and establish tare weights where needed. iv. For plant material, sort and remove any stalks, stems, soil, or items that do not meet the definition of marihuana and would improperly increase the weight of the plant material. 1. Following normal sampling technique, place a representative sample of plant material into a zip-lock bag or other container to be transported to the lab; this is called a core sample. a. Collect enough material to allow analysis in the laboratory for microscopic, color test, and GC/MS analysis. b. Mark the samples that will be used for analysis. 2. If material is wet or moldy, obtain a wet weight if possible; however, note the condition of the evidence on the Notice of Marihuana Core Sampling Sheet and note when an accurate weight cannot be established. 3. Sample wet or moldy material for analysis if possible. d. Inspect and inventory all remaining primary containers. i. If the items appear similar in size and appearance and the contents appear consistent with previous items, take a single representative core sample from each primary container. ii. If the container or its contents do not appear consistent with previously sampled evidence, inventory and collect a core sample as a new piece of evidence. 7. Rewrap and repackage items, label each exhibit, and properly reseal primary containers. 8. Obtain and record the gross exit weight of the primary containers on the Notice of Marihuana Core Sample sheet. 9. Insert the core samples collected from the evidence into a separate evidence bag or container of the submitting agencys choice. a. Place an evidence label on the new evidence bag and enter appropriate information on the label including the unique forensic laboratory number. b. The evidence bag is properly sealed by IFS staff, initialed by both IFS staff and agency witness, and weighed. i. The weight is recorded on the outside of the evidence container by IFS staff.
ii. The evidence bag is released to law enforcement staff to be submitted to IFS at a later date. iii. Submission of these core samples follows the same protocol as all other drug evidence. 10. Close the chain of custody on the primary containers by returning the container back to the property room staff with completion of chain of custody on the Notice of Marihuana Core Sample sheet. 11. The Chemist performing the core sample will typically analyze the case once it is submitted to the Laboratory.
Off-Site Sampling Version 2.0
PHARMACEUTICAL ANALYSIS General Information: 1. Pharmaceutical preparations such as tablets, capsules, patches, and lollipops typically are not quantitated. 2. Identification is performed by Logo Identification and/or GC/MS. Logo Reference Sources: Acceptable logo reference sources include a PDR, Ident-A-drug, Drug ID Bible, a manufacturers website, or other primary reference. Questions about the suitability of a logo reference source should be directed to a supervisor. Analytical Procedure: Pharmaceutical identification falls into two categories: identification. identification by GC/MS and/or visual
1. Visual Identification a. Determine the total weight and number of items to be examined and record on Drug Protocol Worksheet. b. Assess whether the materials appear to be legitimate pharmaceuticals. i. Materials suspected of being clandestine, tampered with, or with incomplete logo are not suitable for Visual Identification. 1. Analyze by another technique such as GC/MS; seek supervisory assistance as needed. ii. Capsules are not typically analyzed solely by Visual Identification. c. Determine the identity of the item(s) to be examined using an acceptable logo reference source: i. Visually examine each item(s) for logo and appearance. ii. Document visual appearance and markings on the Drug Protocol Worksheet. iii. Document the number of item(s) to be examined per logo on the Drug Protocol Worksheet. iv. Obtain a printed copy of the logo identification from an acceptable reference and include in the case file. d. Results Reporting i. When this procedure is solely used to identify a pharmaceutical, the report must clearly state that the tablet or capsule was visually identified as 2. GC/MS a. Determine the total weight and number of items to be examined and record on the Drug Protocol Worksheet. b. Visual Identification i. Perform a Visual Identification as described above if possible. c. GC/MS i. For Visually Identified pharmaceuticals, transfer an amount of sample that is
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory 1 Pharmaceutical Analysis Version 2.0
equivalent to approximately 1 mg of drug to a 15 mL screw cap culture tube. 1. For unknown materials, transfer approximately 100-200 mg of sample to a 15 mL screw cap culture tube. ii. Add 1 mL of methanol. iii. Vortex and/or centrifuge as necessary. iv. Transfer an aliquot to an autosampler vial and inject on the GC/MS using BLANK.M method. v. It may be necessary to concentrate and or run on a splitless method such as LOWDOSE, LSD, etc. d. Results Reporting i. When this procedure is used it must be clearly stated on the report that the pharmaceutical was used for analysis. ii. If a visual identification was performed, the report should also state that the pharmaceutical is manufactured to contain 3. Quantitation a. If quantitation is required, refer to an appropriate method or seek supervisory assistance.
Pharmaceutical Analysis Version 2.0
Drug Analysis Laboratory SCREENING UNKNOWN SAMPLES Principle In most cases, the analyst is able to select appropriate analytical procedures based upon color test results, markings on pharmaceuticals, visual inspection, and/or professional judgment and experience. When traditional methods fail to indicate a testing approach, the following method is used to screen unknown samples and provide guidance for additional testing. Sample Requirements: Powders, solids, liquids, tablets, capsules, residues, and other types of drug evidence are suitable for analysis using this procedure. Equipment: Gas chromatograph-mass spectrometer containing a methylsiloxane capillary column such as an HP-5MS, 30 m x 0.25 mm x 0.25 um film thickness and/or gas chromatograph-flame ionization detector with split/splitless injector containing a methylsiloxane capillary column such as an HP1, 15 m x 0.32 mm x 1.0 um film thickness (see GC/MS and/or GC Methods Notebook in Drug Laboratory). Screw top culture tubes, 15 mL, 16 mm x 125 mm, screw cap with Teflon liner Autosampler vials, 32 mm x 11 mm, Teflon lined seals and glass inserts Vortex mixer Test tubes Pipettes, various Reagents: Methanol, reagent grade or better Safety Precautions: The most common chemical exposure in this type of laboratory is a chemical splash to the skin or eye. Skin, mucous membranes, or eyes which have been splashed with commonly used chemicals or biologicals should be thoroughly washed for at least 15 minutes in cool tap water or eye wash station. Refer to MSDSs for additional chemical information. Report the incident immediately to a supervisor. Seek medical attention as necessary. Methanol is a flammable solvent. Use in a hood or in a well ventilated area. Color Testing:
Screening Unknowns Version 2.0
See the section on Color Testing. Spot tests will usually be run on each item of nonpharmaceutical evidence within one case up to the applicable statutory weight limit. Color testing may be performed on pharmaceutical evidence as applicable. Analytical Approach
Solid
Determine net weight of sample
Liquid
Determine net weight and volume of sample
Residue
Wash sample container with methanol and reserve washing in sample tube.
Perform color test(s)
Perform color test(s) and test pH and solubility as needed.
Run methanol blank by GC/MS using BLANK.M. Run methanol wash of evidence by GC/MS using BLANK.M.
Dissolve arbitrary amount of sample in methanol.
Dissolve 1 part sample to 4 parts methanol.
Run solution by GC/MS using BLANK.M
Run solution by GC/MS using BLANK.M
If no controlled substance is detected or a weak spectra is found run sample on LOWDOSE.M, VIAGRA.M or LSD.M .
Empty autosampler vial back into sample tube and rinse vial with methanolic HCl. If no controlled substance is detected or a weak spectra is found run sample on LOWDOSE.M, VIAGRA.M or LSD.M. If no controlled substance is detected or a weak spectra is found run sample on LOWDOSE.M, VIAGRA.M or LSD.M.
Evaporate to dryness.
Once unknown is identified refer to appropriate method to complete analysis.
Instrument Parameters: See GC Methods and GC/MS Methods Notebooks. Reporting Criteria: 1. The identity of controlled substances determined by GC/MS is reported. a. Other procedures as applicable may be used to quantitate the controlled substance. 2. If no controlled substance is identified it will be so noted on the report. 3. Other types of testing such as IR may also be used in an attempt to identify an unknown substance. 4. Seek supervisory assistance as needed.
WEAK ALKALINE DRUG ANALYSIS Principle of Assay: This method describes the analysis of drugs which are weakly alkaline (weak bases) and/or which are not stable in the presence of strong base. Examples of drugs identified and quantitated by this method are cocaine including cocaine in oil, injectable forms of morphine and hydromorphone, phencyclidine, and chlordiazepoxide. Powders and solids suspected of containing cocaine are typically analyzed using the Cocaine Direct Dilution procedure. Drugs are identified using analytical techniques which provide conclusive identification of the drug. These techniques include gas chromatography/mass spectrometry (GC/MS) and infrared spectroscopy (IR). The weakly basic drug is dissolved in water and the pH of the solution is made weakly basic (approximately pH 8.5) using sodium bicarbonate. In the weakly basic aqueous solution, the weakly basic drug is converted from its ionic salt form to its nonionic free base form. In this form, the drug is quantitatively extracted into n-butyl chloride or chloroform containing phenanthrene as the internal standard. Quantitative analysis is performed on the extracted drug using gas chromatography (GC). Marked pharmaceuticals will usually be identified by logo match and GC/MS without quantitation. However when the penalty group or schedule of a marked pharmaceutical preparation is determined by dosage or preparation, sufficient testing should be performed for placement of the substance in a proper penalty group or schedule. Sample Requirements: Powders, solids, liquids, tablets, capsules, residues, and other types of drug evidence are suitable for analysis using this procedure. Equipment: Gas chromatograph-mass spectrometer containing a methylsiloxane capillary column such as HP-5MS, 30 m x 0.25 mm x 0.25 um film thickness and/or gas chromatograph-flame ionization detector with split/splitless injector containing a methylsiloxane capillary column such as HP-1, 15 m x 0.32 mm x 1.0 um film thickness (see GC/MS and/or GC Methods Notebooks in Drug Laboratory). Screw top culture tubes, 25 mL, 20 mm x 125 mm, screw cap with Teflon liner Screw top culture tubes, 15 mL, 16 mm x 125 mm, screw cap with Teflon liner Volumetric flasks, 10 mL, 25 mL, 2 liter Autosampler vials, 32 mm x 11 mm, Teflon lined seals and glass inserts Vortex mixer Rotator
Dallas County Institute of Forensic Sciences Controlled Substances Laboratory Weak Alkaline Drug Analysis Version 2.0
Centrifuge Test tubes pH paper Pipettes, various Hamilton syringes, various Analytical balance Adjustable 10 mL Dispensette Reagent Bottle Mortar and Pestle Erlenmeyer flask, 50 mL Graduated cylinders, various Reagents: n-Butyl chloride, reagent grade or better Chloroform, reagent grade or better Phenanthrene, reagent grade or better HCl, reagent grade or better Sodium bicarbonate, reagent grade or better Methanol, reagent grade or better Analytical standards Methanolic HCl (5%) Transfer 95 mL of methanol to a 100 mL Erlenmeyer flask then add 5 mL concentrated HCl. Acetonitrile, reagent grade or better Hexane, reagent grade or better Reagent Volume of DI water Volume of HCl
25% HCl 30 mL 10 mL In the hood, place 30 mL deionized water into a 50 mL graduated cylinder and carefully add 10 mL of concentrated HCl using a graduated cylinder. Transfer to a 50 mL Erlenmeyer flask for storage. Safety Precautions: The most common chemical exposure in this type of laboratory is a chemical splash to the skin or eye. Skin, mucous membranes, or eyes which have been splashed with commonly used chemicals or biologicals should be thoroughly washed for at least 15 minutes in cool tap water or eye wash station. Refer to MSDSs for additional chemical information. Report the incident immediately to a supervisor. Seek medical attention as necessary. n-Butyl chloride is flammable. Use in a hood or in a well ventilated area. Do not make contact with skin. Hydrochloric acid is corrosive. Avoid contact with skin, eyes, and mucous membranes. Hexane and methanol are flammable solvents. Use in a hood or in a well ventilated area.
Chloroform is a chlorinated solvent and is considered a human carcinogen. Use in a hood or in a well ventilated area. Avoid contact with skin. Acetonitrile is a suspected teratogen. Use in a hood or in a well ventilated area. Avoid contact with skin. Internal Standard (I.S.) Solution Preparation: Internal Standard Amount of Chemical Final Volume Solvent
0.5 mg/mL phenanthrene 1.00 grams 2.0 L n-butyl chloride Transfer 1.00 grams phenanthrene to a 2.0 L volumetric flask and bring to volume with solvent. Mix by inverting the flask. Calibration Standards: All solutions of stocks and standards are kept in the refrigerator. Allow the solutions to come to room temperature before using them. The concentration, amount, and volume of the calibration stock and calibration standards may vary proportionally depending on the availability of drug, and the expected concentration range of the drug. The following is provided as a guide: Calibration Stock Amount of Drug (as base) Final Volume Solvent
5.0 mg/mL Drug 50.0 mg 10 mL deionized (DI) water Transfer 50.0 mg drug calculated as the free base to a 10 mL volumetric flask and bring to volume with DI water. Mix by inverting the flask. Sonicate as needed. Transfer solution to a 15 mL culture tube for storage. Calibration Procedure: 1. Choose a minimum of three appropriate calibration standards for the calibration curve based on the linear range required for the sample or drug to be analyzed. 2. Using a Hamilton syringe or volumetric pipette, place the appropriate amount of calibration stock into a 15 mL labeled, screw cap culture tube. Add deionized (DI) water to make a total volume of 5.0 mL. Add approximately 200 mg sodium bicarbonate and 5.0 mL internal standard solution. Calibration Standards 0.10 mg/mL 1.00 mg/mL 5.00 mg/mL Amount of Calibration Stock 0.1 mL 1.0 mL 5.0 mL Amount of DI H2O 4.90 mL 4.00 mL 0 mL Volume I.S. solution 5 mL 5 mL 5 mL
3. Cap the tubes and place on a rotator for 5 - 10 minutes.

4. Centrifuge for 5 - 10 minutes on medium setting. 5. Transfer an aliquot of the organic layer to an autosampler vial. Note: n-butyl chloride will be on the top; chloroform on the bottom. 6. Inject the extracted calibration standards into the GC using the appropriate instrument method. 7. Determine if the data are acceptable using the instructions found in the Calibration Curve/Response Factor Log. An acceptable calibration curve has a correlation coefficient, r2 > 0.995. If the curve is acceptable, file it in the Calibration Curve/Response Factor Log located in the Drug Laboratory. If not, seek supervisory assistance as needed. Quality Control Standard: Ideally the quality control standard will be made from a different lot number or manufacturer than the calibration stock. At a minimum, it should be made from a different stock solution. A quality control standard must be successfully run on each day this procedure is used. The concentration, amount, and volume may vary proportionally depending on the availability of drug. The following is provided as a guide: QC Stock Amount of Drug (as base) Final Volume Solvent
2.0 mg/mL Drug 50.0 mg 25 mL deionized (DI) water Transfer 50.0 mg drug calculated as the free base to a 25 mL volumetric flask and bring to volume with deionized water. Mix by inverting the flask. Sonicate as needed. Transfer solution to 25 mL culture tube for storage. Quality Control Procedure: 1. Using a volumetric pipette, place 2.5 mL of quality control stock and 2.5 mL deionized water into a 15 mL labeled, screw cap culture tube. Add approximately 200 mg sodium bicarbonate and 5.0 mL internal standard solution. QC Standard 1.0 mg/mL Drug Amount of QC Stock 2.5 mL Amount of DI water 2.5 mL Volume of I.S. solution 5.0 mL
2. Cap the tube and place on rotator for 5 - 10 minutes. 3. Centrifuge for 5 - 10 minutes on medium setting. 4. Transfer an aliquot of the organic layer to an autosampler vial. Note: n-butyl chloride will be on the top and chloroform on the bottom of the respective water layers. 5. Inject the extracted quality control sample into the GC using the appropriate instrument method. Evaluation Criteria:
1. If the QC sample is within the set limits of +/- 5% of the target concentration, proceed with analysis. 2. If the QC sample is not within the set limits of +/- 5% of the target concentration, additional action must be taken to obtain an acceptable QC result such as re-running QC, re-extracting and re-running QC, making new stock with re-extraction and reanalysis, recalibrating (run curve), and/or consulting a supervisor. Color Testing: See the section on Color Testing. Spot tests will usually be run on each item of nonpharmaceutical evidence within one case up to the applicable statutory weight limit. Color testing may be performed on pharmaceutical evidence as applicable. Analytical Procedure: 1) Powdered Samples, Hard Chunky Powders, and Other Solids: a) Obtain the total weight of the exhibit to be analyzed and record on the Drug Protocol Worksheet. b) Create a representative specimen by taking a sample from many or all of the solid pieces in the exhibit. If powder is also present, remove a sample and add to the material scraped from the solid pieces. Pulverize the resulting composite sample and analyze as described. c) GC/MS Identification i) Transfer approximately 10 mg of material to 1 mL methanol in a test tube. (1) The analyst will vary the amount of material and methanol as needed to obtain an acceptable mass spectrum. ii) Vortex and centrifuge as necessary. iii) Transfer an aliquot to an autosampler vial and inject on the GC/MS using BLANK.M method. d) GC Quantitation i) Weigh approximately 10 - 50 mg of the material to be analyzed, and record the weight on the Drug Protocol Worksheet. ii) Transfer sample to a 15 ml labeled, screw cap culture tube. iii) Add 5.0 mL of deionized water and approximately 200 mg sodium bicarbonate to the culture tube. iv) Add 5.0 mL of internal standard solution. v) Cap the tube and place on rotator for 5 - 10 minutes. vi) Centrifuge for 5 - 10 minutes on medium setting. vii) Transfer an aliquot of the organic layer to an autosampler vial. Note: n-butyl chloride will be on top; chloroform on the bottom. viii) Inject extracted sample into GC using the appropriate instrument method and calculate the results. 2) Tablets, Capsules, and Other Pharmaceutical Preparations: a) Obtain the total weight of the exhibit to be analyzed and record on the Drug Protocol
b)
c) d)
e)
Worksheet. Create a representative sample for analysis; grind to a powder if necessary. i) Capsules typically a composite sample is used for analysis. ii) Tablets typically one tablet is selected for analysis. Logo Identification i) Refer to Pharmaceutical Analysis Procedure. GC/MS Identification i) Transfer an amount of sample that is equivalent to approximately 1 mg of drug to approximately 1 mL methanol in a test tube. (1) The analyst will vary the amount of material and methanol as needed to obtain an acceptable mass spectrum. ii) Vortex and centrifuge as necessary. iii) Transfer an aliquot to an autosampler vial and inject on the GC/MS using BLANK.M method. GC Quantitation i) Weigh approximately 10 50 mg of the material to be analyzed and record the weight on the Drug Protocol Worksheet. ii) Transfer sample to a 15 mL labeled screw cap culture tube. iii) Add 1.0 - 5.0 mL of deionized water, approximately 200 mg sodium bicarbonate, and 1.0-5.0 mL of internal standard solution. iv) Cap the tube and place on rotator for 5 - 10 minutes. v) Centrifuge for 5 - 10 minutes on medium setting. vi) Transfer an aliquot of the organic layer to an autosampler vial. Note: n-butyl chloride will be on the top and chloroform on the bottom. vii) Inject the extracted sample into the GC using the appropriate instrument method and calculate the results.
3) Liquid Samples: a) Liquid samples may be analyzed by weight or by volume. If the sample is to be analyzed by volume, a known volume of the liquid must be weighed to determine density. b) If the volume of liquid is small, it should be treated as a residue; see below. c) Obtain the total weight of liquid in the exhibit and record on the Drug Protocol Worksheet. If the liquid is to be analyzed by volume, record the total volume on Drug Protocol Worksheet. d) GC/MS Identification i) Place approximately 1 part sample to 4 parts methanol in a 15 mL screw cap culture tube. Mix well before sampling. (1) The analyst will vary the amount of material and methanol as needed to obtain an acceptable mass spectrum. ii) Transfer an aliquot to an autosampler vial. Analyze by GC/MS using BLANK.M method. e) GC Quantitation i) Transfer a known volume or weight of the sample to a 15 mL labeled screw cap culture tube. ii) If the liquid is aqueous, proceed to step iii. If the composition of the liquid is
unknown or organic, add three drops of methanolic HCl and evaporate the sample to dryness or to constant volume. iii) Based on the estimated drug concentration of the material, determine the extraction volume. The volumes of water and extraction solvent should be equal: add 1.0 5.0 mL of deionized water, add approximately 200 mg sodium bicarbonate, and 1.0 - 5.0 mL internal standard solution. iv) Cap the tube and place on rotator for 5 - 10 minutes. v) Centrifuge for 5 -10 minutes on medium setting. vi) Transfer an aliquot of the organic layer to an autosampler vial. Note: n-butyl chloride will be on the top and chloroform on the bottom. vii) Inject the extracted sample into GC using appropriate instrument method and calculate the results. 4) Residue Samples: a) Carefully rinse the residue into a container using methanol. b) GC/MS Identification i) Run a methanol blank before each residue sample using DRUGANA.M or BLANK.M methods; place in case file. ii) Place an aliquot of the methanol washing into an autosampler vial. Analyze by GC/MS using BLANK.M method. c) GC Quantitation i) Place the sample/methanol mixture in a 15 mL screw cap culture tube, add 1-3 drops of methanolic HCl, and evaporate to dryness. ii) Based on the estimated drug concentration of the material, determine the extraction volume. Add the same amount of water and extraction solvent: add 1.0 5.0 mL of deionized water to residue, approximately 200 mg sodium bicarbonate, and 1.0 - 5.0 mL internal standard solution. iii) Cap the tube and place on a rotator for 5 - 10 minutes. iv) Centrifuge for 5 - 10 minutes on medium setting. v) Run a methanol blank before each residue sample using the same instrument method used for the sample; place in case file. vi) Transfer an aliquot of the organic layer to an autosampler vial. Note: n-butyl chloride will be on the top and chloroform on the bottom. vii) Inject the extracted sample into GC using appropriate instrument method and calculate the results. If it is necessary to dilute the residue, return to the original washing for dilution. d) When analysis is complete return any extracted sample remaining in the autosampler vial to the original culture tube used for the extraction. i) Remove and discard the aqueous layer from the culture tube. Add 1-3 drops of methanolic HCl to the extracted sample to convert the drug to its more stable hydrochloride salt form. ii) Take the extracted sample to residue in a laboratory fume hood. Close with a screw cap. iii) Label the tube added by laboratory, include FL#, initials, and place this tube in the evidence bag along with the evidence.
5) Phencyclidine (PCP) Analysis a) Cigarette Samples i) Obtain the total weight of the cigarette(s) in the exhibit and record on the Drug Protocol Worksheet. ii) GC Quantitation (1) Weigh approximately 100 - 150 mg of the cigarette(s) to be analyzed. (2) Determine weight of the material and record on Drug Protocol Worksheet. (3) Finely cut the cigarette sample into pieces. (4) Transfer cut sample to a 15 mL labeled culture tube. (5) Add a 1.0 5.0 mL of water and approximately 200 mg sodium bicarbonate to the culture tube. (6) Add 1.0 - 5.0 mL of internal standard. (7) Cap the tube and place on rotator for 5 - 10 minutes. (8) Centrifuge for 5 - 10 minutes on medium setting. (9) Transfer an aliquot of the organic layer to an autosampler vial. (10) Inject extracted sample into GC using PCP.M method and calculate the results. iii) GC/MS Identification (1) Analyze a separate sample using a second aliquot by GC/MS using BLANK.M method. b) PCP in Solvent i) Obtain the total weight of liquid in the exhibit to be analyzed and record on the Drug Protocol Worksheet. ii) GC Quantitation (1) Pipet approximately 50 - 200 uL of sample into a 15 mL labeled screw cap culture tube. (2) Determine weight of the material to be analyzed and record on Drug Protocol Worksheet. (3) Add a 1.0 5.0 mL of water and approximately 200 mg sodium bicarbonate to the culture tube. (4) Add 1.0 - 5.0 mL of internal standard. (5) Cap the tube and place on rotator for 5 - 10 minutes. (6) Centrifuge for 5 - 10 minutes on medium setting. (7) Transfer an aliquot of the organic layer to an autosampler vial. (8) Inject extracted sample into GC using PCP.M method and calculate the results. iii) GC/MS Identification (1) Analyze a separate sample by GC/MS using a second sample aliquot. 6) Analysis of Cocaine In Oil a) Obtain the total volume and weight of liquid in the exhibit and record on the Drug Protocol Worksheet. b) GC/MS Identification i) Place approximately 1 mL of material to be analyzed in a 15 mL screw cap culture tube, add approximately 1 mL of hexane, mix well by vortexing. (1) The analyst will vary the amount of material and methanol as needed to obtain an
acceptable mass spectrum. ii) Add approximately 2 mL acetonitrile. iii) Cap the tube and place on rotator for 5 10 minutes. iv) Centrifuge 5 10 minutes. v) Transfer an aliquot of the acetonitrile layer (bottom) to an autosampler vial. Analyze by GC/MS using BLANK.M method. c) GC Quantitation i) Place approximately 1 - 2 mL of material to be analyzed into a 15 mL labeled screw cap culture tube. ii) Determine weight of material to be analyzed and record on Drug Protocol Worksheet. iii) Add an equal volume of water followed by 1-3 drops of 25% HCl. iv) Rotate 5 10 minutes. v) Centrifuge for 5 - 10 minutes on medium setting. vi) Transfer aqueous layer to a clean 15 mL culture tube. vii) Add approximately 200 mg sodium bicarbonate. Test with pH paper to make sure solution is neutral before proceeding. viii) Add an equal volume of n-butyl chloride with internal standard and record on Drug Protocol Worksheet. ix) Rotate 5 10 minutes. Centrifuge for 5 - 10 minutes on medium setting. x) Transfer an aliquot of the organic layer to an autosampler vial. xi) Inject extracted sample into GC using COC.M method and calculate the results. Instrument Parameters: See GC and GC/MS Methods Notebooks located in Drug Laboratory. Reporting Criteria: 1. Report both the quantity and the identity of the drug when the concentration of the drug is within +/- 20% of the calibration curve on the GC and the GC/MS is complete. It may be necessary to dilute extracted samples to bring them within the curve range. 2. Report the amount of the drug as insufficient for quantitation when the drug concentration is below 20% of the calibration curve on the GC and the GC/MS is complete. 3. For PCP cigarette exhibits, report the amount of PCP in the aliquot analyzed, and the total weight of the cigarette aliquot tested, and the total weight of the material in the exhibit. Cigarettes are typically not uniformly coated with PCP; therefore, calculation of the amount of PCP in the entire cigarette may not be accurate unless the entire cigarette was used for analysis. Calculations: 1. Dilutions a. In any situation in which a dilution is made, multiply the concentration of drug (mg/mL) by the dilution factor. A notation relating to how dilutions were made
2.
3.
4.
5.
must be made in the case file. Materials Analyzed by Volume: a. concentration of drug (mg/mL) x volume of internal standard solution used for extraction (mL) x total exhibit volume/extracted sample volume = mg of drug in the exhibit as the free base Materials Analyzed by Weight: a. concentration of drug (mg/mL) x volume of internal standard solution used for extraction (mL) x total exhibit weight (mg)/extracted sample weight (mg) = mg of drug in the exhibit as the free base Percent drug in the exhibit: a. mg of drug in the exhibit/total weight of the exhibit x 100 = % of drug in the exhibit Amount drug in mg/100 mL: a. mg / 100 mL = mg (drug) X 100 Total Volume of exhibit
10
Weak Alkaline Drug Analysis Version 2.0
Training Notes: Weak Alkaline Procedure 1. If GC/MS results are negative when run on BLANK.M, run on alternative GC/MS methods such as LOWDOSE.M, LSD.M, etc. until satisfied that no controlled substance is present. 2. A sonicator may be used to bring materials into solution. Care should be used regarding the length of time materials are left in the sonicator since heat may decompose some drugs. Typically, a sonication time of 5 minutes or less is used. 3. The toxicity of n-butyl chloride is less than that of chloroform; therefore, where possible, n-butyl chloride is the extraction solvent of choice. However, extraction efficiency of some drugs is low or variable in this solvent, and in this case, chloroform is used. 4. To determine whether cocaine is present in the free base or hydrochloride form, refer to the Color Testing procedure for presumptive salt/base determination and to the Infrared Spectroscopy procedure for conclusive identification of salt/base form. 5. Diphenhydramine and ketamine co-elute when run by GCMS using BLANK.M. Use SLOWRAMP.M method if there is evidence of the mixture. 6. Calculation of drug as free base: Option 1 To calculate the amount of drug as a salt required to produce a target amount of free base, use the following equation: MW drug (salt) / MW drug (free base) X target amount of free base Note: Occasionally two molecules of drug are included in the salt form, for example morphine sulfate. Check the chemical formula prior to making the standard. In the case of morphine sulfate, the molecular weight of the salt should be divided by 2 prior to using the formula above. Example: Cocaine HCl (C17H21NO4, HCl) Cocaine (base) MW = 339.8 MW = 303.4
339.8 X 50 mg = 55.9 mg cocaine hydrochloride 303.4 7. Calculation of drug as free base: Option 2 Alternatively, to calculate the percentage of free base in the salt molecule to produce a target amount, use the following equation: Example: Cocaine HCl (C17H21NO4, HCl) Cocaine (base) % base in salt = 303.4 = 0.893 339.8 Amount of salt needed = 50.0 mg = 55.9 mg cocaine hydrochloride 0.893
MW = 339.8 MW = 303.4
11

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