Ms, MILES Technicon RA® Systems Technicon RA® SYSTEMS METHODS MANUAL PUBLICATION NO. TH9-1589-01 / MAY 1994 This publication supersedes Publication No. TH9-1589-00, dated April 1991MILES. Introduction ‘System Applicability: TECHNICON RA® Systems METHODS INTRODUCTION Each TECHNICON RA® system (TECHNICON RA-S00° system, TECHNICON RA-1000* system, TECHNICON RA-XT™ system or TECHNICON RA-2000™ system) is a computer-contolled, random access clinical chemistry analyzer. Stored ‘chemistry parameters direct tne ‘analytical process. ‘The TECHNICON RA-500 system has a total capability of 62 Technicon-defined and user-defined methods of which 32 methods may be immunoassay chemistries. ‘The TECHNICON RA-1000 system has a total capability Ff 62 Techricon-tefined and user-defined methods of which 32 methods may be immunoassay chemistries Additionally, five (5) lon-selactive Electrode (ISE) methods are resident in the system software. ‘The TECHNICON RAXT system has a total capability of 69 Technicon-defined methods and 8 user-defined ‘methods of which 40 may be immunoassay chemistries. Adgitionally, five (6) lor-selective Electrode (ISE) ‘methods are resident in the system software. ‘The TECHNICON AA-2000 system has a total capability of 255 methods, of which 128 are Tochnicon-defined and 127 are user-defined. Of the 255, 100 may be immunoassays, and five (5) are lon-selective Electrode (SE) methods. Information regarding the use of the TECHNICON RA system's reagents on the TECHNICON ASSIST® clinical analyzer or the TECHNION RA-100™ system, may be found in the appropriate application guide. AA Getiled listing of the current Technicon-defined and User-defined chemistries may be found in Table 2 on ages INTRO-7 and INTRO-8 of this section. STATEMENT OF CONTENT This Manual contains specific, msthod-related information and data for the Technicon methods pertormed on TECHNICON RA systems. ‘The procedures necessary for the preparation, use, and storage of TECHNICON RA systems reagents and ‘calibrators are provided in the applicable method sheet. NOTICE ‘Sota earerans pearance ane heway stasis ne 2S ees eae Eeaaeeesrareae marae a St SSN ES, one SAMPLE COLLECTION AND PREPARATION Patient samples should be prepared according to the protocol established by the user's laboratory’ unless otherwise specified in the individual method sheet. A full discussion of sample collection and preparation is beyond the scope of this publication, The subject is covered in detail in recent texts by Kaplan and Pesce! and Tietz2. In addition, the National Committee for Clinical Laboratory Standards (NCCLS) has published two documents on sample collection.> In general, plasma and serum should be separated trom ‘cells within two (2) hours alter collection. i samples cannot be assayed immediately after separation. they should be stored in stoppered containers at 4 °C to 8°C. However, some analytes are adversely affected by low temperatures. Glucose test values may decrease upon prolonged contact with erythrocytes due to the presence of ‘ycolytic enzymes. The use of a preservative such as fluoride should be considered when unseparated samples could stand at room temperature for two (2) hours or more bafore being processed. Samples fram leukemic patients also may require preservatives due to the high metabolic activity of white cells. Special storage conditions. such as freezing, protection from Girect exposure to light, or the addition of a preservative for other analytes will be noted in the method sheets where applicable. During sample collection and preparation. hemolysis of erythrocytes should be avoided. The elfect of hemolysis on specific methods is described in the interference section of the applicable method when i is significant. The timely separation of serum from cells is especially important for certain assays. such as lactate dehydrogenase, magnesium, and potassium. Blanking and other analytical strategies can offen reduce the interference by hemoglobin pigments, but cannot correct the elevation of serum levels of analytes normally found in higher concentrations in erythrocytes. These chemistries include potassium. magnesium, and lactate dehydrogenase, ‘The blood sample should be collected in a commercially available collection tube using routine venipuncture techniques. Samples should be processed in a manner that will prevent the introduction of ciats into the system. For this purpose, the TECHNICON SeraCiear® serum filtering device (Prod. No. 510-40SSPO1) is recommended for use as follows mites JA Miles tn Diagnosties Division ‘Tarrjtown, NY 10591-5097 USA. INTROINTRODUCTION USE OF TECHNICON SeraClear® FILTERING DEVICE WARNING- POTENTIAL BIOHAZARDOUS MATERIAL men Sond asma,o swum shed be nanded caucus as SES marr ecaang go aay ent Sscause ro test metod can aftr comoite asuronce tat oman inruneelsercy Vos (49, Daas 8 wo tet lectus apes re ssa, eemens haute hari othe Sesatty Let 2 as recommend fr ary pote ecto human ‘erm or ieod spcinen the Goss fr ease Gonvointonel inaeaten of Heath manus! “Bovaley at ‘icbogee! and Boned! abaratres 88s 1. After drawing the specimen trom the patient Perform identlicaion procedures consistant with Yyour laboratory protocol. Place the collection tube im an upright vertical position and allow the Specimen to stand undisturbed for @ pend of thity (60) minutes prior to centhtugation. 2. Before placing the specimen in the centrifuge, carefully rim the collection tube to free the clot om the glass. Perform this procedure carefully so that the cells are not nemolyzed, 3. For optimum separatior. the specimen should be centrifuged for twenty (20) minutes at a speed that ‘raduces @ relative centrifugal force (RCF) of 2200 x gravity (g). Consult centrifuge manutacturers Instructions for further information. It the TECHNICON SeraClear titering device 18 not used proceed to step 6. IMPORTANT Belore using the TECHNIGON SeraClear serum fmonng deve inspect carat to venty tat te vent Mole is clear and twat he wiper not bent SF astones. in oer avad paseo exponre TECHNICON seraclear serum ‘itenng. deuce Inathas a bocked vant hole, ora Cont wipe. DANGER Sete mnsorung 2 TECHNICON SeraCloar serurn fiteeng Gevece mio a specimen tute a5 srected ne following’ siga.carelully check the TECHNIGON Seracicar sorum therng deve Io be Sure ns ternal fers present. A missing hie Bemis the 'samcie 10 nse. too fancy me the TECHNIGON’ SeraClear serum ‘tena ‘weviee and may lead 19 sample ‘selage anc posse Sentamination of persannel andor laboratory inswuments.” Goad. laboratory practice “3136 fequires inatguring iasection, the TECHNICON SeraCiear sorum ttonng cowed De pomed away tom the user's lace, Technicon recommends the use of appropriate eye and hand protection when ising this device. 4. Caretuly inser the TECHNICON SeraCtearfitering Gevice into the mouth of the collection tube. During Insertion. keep the filtering device. straight and directly in line with the Jong axis of the collection tube, This will prevent the wiper from being bent. 5. Slowly press the fitering device into the collection lube until the bottom of the filter is positioned approxmately 0.5 mm above the packed ved cells lt the device ts pushed past the point specified. he Packed cells may be disturbed, and pass through the fter IntRO-2 REVISED: 6. Carefully. decant the serum from the original collection tube into a clean TECHNICON RA systems sample cup. Make certain that no clots are decanted into the clean cup. OPERATION All information pertaining to the operation of TECHNICON RA systems is included in the applicable TECHNICON RA system Reference or Operation ‘Manual provided with your system. CALCULATION OF RESULTS TECHNICON RA systems automatically perform all calculations that are necessary for abtaining final results. ‘The system prints out the results for each sample in concentration units. The derivation of results from raw absorbance data is explained in the “Calculation of Results" section of the applicable TECHNICON RA system Reference or Operation Manval INTERPRETATION OF RESULTS System operators and laboratory supervisors are responsible for operating and maintaining Technicon products in accordance mith the procecures described in the applicable Product Labeling (Operator Manuals. Package Inserts, Reference Manuals. Bulletins. efc) and for determining that these products conform to the applicable performance claims, |, under these prescribed conditions of operation and maintenance, an aberrant or abnormal result. as defined by the laboratory protocol. occurs. laboratory personnel should first make certain that the system is performing and is being operated in accordance with the Product Labeling: then follow the laboratory protocol for advising the clinician of a result that appears to have deviated from the norms established by the laboratory. Technicon makes every effort to identify in the Troubleshooting, Maintenance, and Bibliography sections of the Product Labeling. the potential effects caused by instruments, reagents. or endogenous Substances, including known interferences on patient results. However, its impossible to provide information whieh would be ‘applicable to every clinical situation This information ts provided in lieu of attempting 10 define the need for additional testing which involves: medical judgement in each individual case. The decision whether or nat tO report @ diagnostic result rests with the laboratory Technicon products do not make diagnoses on patients, ‘Technicon intends its diagnostic products (systems reagents, sofware, hardware, efc) 10 be used 10 collect ata rellecting the patients chemical, hematological, oF {immunological status at 2 certain point in me. Such data must be used in conjunction with other diagnostic information and with the attending physician's evaluation Of the patient's condition to arrive at a diagnosis and a Clinwcal course of veatment, Any malfunction of a Technicon diagnostic product (i.e failure to meet a performance specification or otherwise to perform as intended) should be appropriately addressed by the laboratory personnel. Various Sections of the Product Labeling previously noted address malfunctions and their possible effect on ‘resultsINTRODUCTION TECHNICON RA SYSTEM METHODS The individual sections that follow deal singularly with the clinical method types that have been reprogrammed into TECHNICON RA systems. For example. the section containing amylase, provides information relevant to the zero-order determination a it 's performed on TECHNICON RA systems. The section Containing glucose. provides information applicable 10 performance of endpoint methods. Each method is tabbed with the name of the analyte to which it applies. The methods are presented in alphabetical order and in a consistent format under the following headings: Intended Use This paragraph contains @ brief statement of intended tse for the method as a in wiro diagnostic procedure for Quanttating analytes in human serum and in some cases plasma and urine, Summary and Explanation This paragraph contains a short istry of the method with pertinent references to previous ‘lated methods Principles of the Procedure This paragraph provides a physical, chemical, and biological éxpianation of the method and a description cf the chemical reactions, whera applicable. Reagents All data regarding the reagents used to perform the ‘method are discussed in this paragraph Throughout ihis Section three words are used to signal the presence of hazards associated with the use and handling of reagents. These words are ~ in descending Order of severity - DANGER! WARNING! and cauTion: The following briefly explains the significance of each of these heacings: DANGER! Signals tno presence of 3 hazarc that 1s ie tveatenng. “thie hazare can result Worn tne Dresence ol, the handing at. or the use Ol vbstances tar are monly tou. exvemely cor-oeve, Grenverely tammabie WARNING! Sinats ne oeserce of a hazard mals myraus but 01 le treatenng_ uncer arénary crcumatances, This nazara can result rom tie beesence oe he hanoing of oF he ube of aybsarces such 09 tox Sous o Gases, wrtans: or ammabie materals CAUTION! Signals te oresence of @ hazard that could cause ness, burns son feastons ate The hazard Could result rom the presence ofthe handing at of te Use of Substances sucn as loud iqude OF gases, iriais. or fanmatie mater, ‘The customer must’ maintain a constant awareness of {he hezarcs and warnings associated min all reagents, Where necessary. aporooniate safety equipment should be used (eg. eve protection, gloves” iab coats) Calibrator and Control Products Table 1 lists the rocucts currently availabe for use with the TECHNICON RA systems Table 1 TECHNICON CALIBRATOR AND CONTROLS Pnopucr PRODUCT WaME var oF 708-1291-82 lrecHNICON SETpoint™ or (TECHN weamt fros-191-00 4 : ECRNICON TESTpomn To To9-1200-62 OU TSTpGnT fra vo mL Taare |TECHICON TESTpomt= > mee |Assayed Chemusiry Controt 2 panied ITECHNICON TESTpoint™ ae 03.3228.01 |Ugena Conte an steve "zor |TEGHMICON SETpomnt™ ae bonniai |CO, CatbratorDituent 2eatme s70 1 [TECHNICON SeTpo1ai= vam 7009174-0" Hy Cafprator GE va77001 | TECANICON SETpoint™ van 703-1770-0% lanstyoue Standards Bao ryaszar [TECHNICON SeTpoiat= vem Tes-s0n2- |Theophyline Standards s TECHNICON SeTpoint™ 103-2050.01 |Prenmbarstlbrenyon orn Standarss 703-1556-01 [Speate Proven Relearce Seum [oa 22 AU 1045-0) |TECHNICON SETpoint™ ae 703-1945:01 |c‘Rescwe Proton Calibrator 7 ITECHNICON TESTpoint™ Foe 09.2075.01 |Seeone Proven Convale Ba tees) Foter to the current Price List. applicable method sheet, ‘of package insert for information concerning these and ‘other Calibration and Control procucts Storage and Stability Reagent storage and expiration dates are printed on the product label. The on-system stabiity statod in. the method sheet raoresents the mmnimumm stable period Yor the reagent. The user however, may elect to use ‘reagents for a longer perod of time if resus tom the ty Contiol Program indicate acceptable Sample Handling Far 2 detailed explanation of sample handling refer to pages INTRO-1 and INTRO-2 of this manual under the headings of “Sample Collection Preparation.” Refer to the applicable TECHNICON RA systems chemistry method sheets for details any special sample handling requirements or dilutions. Procedure The procedures for performing TECHNICON RA systems methods are located in the Operation Secton of the Reference Manual for the applicable TECHNICON RA system Assay Parameters “The assay parameters for the TECHNICON RA 2ystems ‘memoas are preprogrammed into each TECHNICON RA system unless otnermise indicated For the complete 1IS concerning these parameters. re'er to the anplicaole Chemistry Program Data Sheet supplied with each method ‘Avalabe lor use outa Nowh Amenca REVISED: Aon 1991 IntRO-3INTRODUCTION Quality Controt ‘Technicon recommends that TECHNICON RA systems be controlled using commercially prepared control material. For complete details conceming the use of Qualty conl matoals refer to te chemisey metres sheets Calibration This paragraph specifies the procedure for and the frequency of calibration for each method and the ‘material to be used. where appropriate. Results For complete details concerning the explanations for the Calculations of endpoint, frstrorder, zero-order, quadratic te, and blanked zero-order methods refer 10 the TECHNICON RA system Reference Manual Section Il ‘Calculation of Resutts.” Endpoint and First-Order Rate Methods During installation of the system, all endpoint and first-order rate metnods are to be calibrated using TECHNICON SETpoint Calibrator (Prod, No, T03-1291- 62 or T03-1281-D0).Thereatter. cecalibration must be performed after any one ofthe following changes: © Reagent lot ‘© Colorimeter component '® Hydraulic component affecting sample- torreagent volume ratio ‘i \n addition, Quality Contro! (QC) results should be monitored dally. f GC limits are not met. appropriate Corrective measures, incliding recalibration, should be implemented before reporting patient results, The following methods are examples of endpoint methods Albumin Glucose. Tota! Bileubin Calcium Direct Bilirubin Total Protein Cholesterol Inorganic Phosphorus Triglycerides Chioriae Magnesium Une Acid Carbon Dioxide (Colorimetric) The following methods are examples of first-order rate methods Creatinine Urea Nitrogen Zero-order Rate Methods Zero-order rate methods are factory calibrated and do ot require routine calipration, Subsequent recaibration must be performed after any one of the following ‘changes: ‘© Colorimeter component © Hydraulic component affecting sample- torreagent volume ratio The following methods are examples of zero-order rate methods: Alkaline Phosphatase Alanine Aminotansterase Aspartate Aminotransterase Amylase Quadratic Rate Methods Quadratic methods incude simple and fitted immunoassay) types. Typically. immunoassays utilize multipoint calibrations using several types of avalaole algorithms, Quadratic rate methods should be calibrated by the user after installation of the system. Thereafter fecalibration must be performed alter one of the following changes Creatine Kinase y-Glutamyltransterase Cactate Dehydrogenase nTRO: © Reagent fot ‘© Colorimeter component @ Hycraulic component affecting sample- to-reagent volume rato The following methods are examples of quadratic rate ‘methods: Gentamicin Theophylline Phenobarbital Thyroxine (Te) Phenytoia Tobramycin Blanked Zero-order Method (Blanked Rate) ‘Gurrenty, there is only one chemistry cesident in the TECHNIGON RA systems using this method type. This is the Creatine Kinase-2 method. Limitations of the Procedure The section describes if there are any substances, conditions, and patient states that have been reported to Cause physiclogical changes in the analyte Concentration Expected Values The expected valves were calculated using the correlation statistics contained in each method. Ranges fof expected values can vary depending on age. sex. iet, location, etc: therefore, users should evaluate their usual population to determine their own laboratory specific expected ranges. Performance Characteristics ‘This section provides technical information related to method performance characteristics, such as imprecision, correlation, and linearity. Performance observations were gathered over’a period (of several months. A description of each statistical parameter and the protocol followed to compute that parameter are Contained in the paragraphs that follow imprecision imprecision is detined as the degree of variability among repeated independent: measufements of the same sample. \Within-run imoracision refers to the reproducibility of the method when samples are assayed within a single worklist. Total imprecision extends the withinrun estimates to incluge Between run and between day components of vanation imprecision estimates were computed at specified analyte levels using human serum pools and contra! materials in a protocol similar to that recommended in the NCCLS document EP5-P, Proposed Guidelines for User Evaluation of Precision Performance of Cimical Chemistry Dewces * All imprecision data were obtained over several calibration periods using a single lot of individual Feagents and calibrator Precision was evaluated on at least two systems per method. REVISED: pr 1991INTRODUCTION ‘The imprecision values for each method, reported on each of their respective method sheets, are point estimates of average imprecision that can be expected on the TECHNICON RA system. Since this is a point estimate, any single TECHNICON RA system tested with a similar protocol could yield a different estimate of imprecision and yet not be signiicantly different trom the published average imprecision. ‘The NCCLS cocument, EPS:P, describes a statistical test (Chi Square) for comparing an instrument's performance and the expected performance. A user ‘should be able to obtain imprecision data which pass the (Chi Square test at the 85% confidence level. Correlation Data Correlation data were determined where the Performance of TECHNICON RA systems were ‘compared with the performance of a relerence method ‘or a comparative system, where applicabie. Regression statistics were computed using human serum samples in a protocol similar to that ecommended in the NCCLS document EPSP, User Comparison of Quantitative Clinical Laboratory Methods Using Patient Samples. Regression data in each method indicate the linear least squares fit between TECHNICON RA systems (y) and the comparative systemmethod (x). Linearity ‘The linear range of response of each method was studied over the claimed range of measurement using at least five equally spaced leve's of a human secum pool. Each level was assayed multple times and a least squares line or other approprate statical fool was sed fo fit the dala. “Unless ‘stated otherwise, the ‘maximum eviaion rom the ited line was less than 5% atthe upper lt of ean. Occasionally. some specimens may exceed the linear range of the method and produce error flags of messages. The details of these flags and messages is available in the Reference Manual for each of the TECHNICON RA systems. Specimens that produce these errors should be diluted and reassayed according to directions available.n the specific method sheets Autodilution and Extended Linearity on the TECHNICON RA-2000 System An “Autodilution” disposition is available on the TECHNICON RA-2000 system as an option for the ‘operator. This option allows the operator, upon Review and Edit. to rerun the sample ubiizing a smaller alternate Sample volume in order to generate a resull that is greater than the (neat) range high of the method. Depending on the method, linearity can be extended, in many cases. to several times the (neal) range nigh parameter. Generally, the approximate theoretical linearity for each method is simply the ratio of the (neat) ‘% sample volume divided by the % alternate sample volume and that result multiplied by the (neat) range high. Technicon has tested the methods indicated below and has validated this extended linearity to the value listed. and not necessarily to the theoretical limit, ‘which in many cases, has no clinical lly It a result exceeds this new “validation range high” using the alternate sample volume. a message “EXCEEDS VAL ANG" will be printed along with the result on the 40-column instrument printer, or a “V" listed in the TECHNICON RA-2000 system Workstation ‘along with the result. If result, belore being multiplied by the auto-dilution factor of the software, would generate a result greater than the range high of the method, a result iS printed along with the message: “EXCEEDS HIGH RING" on the 40-column instrument printer or listed with 2 "G" on the Workstation, with a Printed numerical value. A manual dilution of the Sample 1s then recommended, as described in its respective method sheet. if a method has no alternate ‘sample volume listed, the chart indicates “N/A” for “Not Available” or Not Applicable” and in those cases. autodilution cannot be used. Evaluations have Gemonsirated that certain sample types andor dilutionimatrix effects could corrupt the result if ‘utediiution was invoked. This phenomena is also evident wnen these kinds of samples are manually iluted with water. The results obtained ‘om those ‘manually ciluted samples could be adjusted, as needed, ‘based an empirical data generated at the system site. 10 produce usable values, For a complete listing of the ‘chemistry linearities and the validation range high values refer to Table 3 on INTRO-9 and INTRO-16. Sensitivity Sensitivity. measured in absorbance (A) per unit of concentration for metabolites. and in Alminute per UiL for enzymes, is listed for each method. For immunoassays, sensitviy 1s typically given in units ‘of mAvmin of the concentration of the drug studied. For example, the phenytoin sensitivity is calculated as mAimin per ugml. Additionally, sensitivity varies with temperature. WASTE DISPOSAL REQUIREMENTS Laws and regulations enacted to protect the environment ‘and to encourage resource conservation. require the Gieposal of hazardous wastes in a species manner Since some of the wastes from TECHNICON RA systems can be classiied as hazardous wastes. it is, ‘essential that the laboratory take appropriate steps 10 etermine the laws and regulations applicable to ther ‘disposal and to effect compliance therew Itis necessary for you to sample your effluent in order to evaluate compliance with applicable regulations. The laboratory should contact a local licensed hazardous ‘waste disposal firm for assistance. The principal wastes associated with the use of TECHNICON RA systems are used reagent cuvettes. reaction trays, sample containers, and system waste ‘CAUTION Oo not incinerate the rescdon tas Lnoer Sample containers with human specimens. control materals, and all reagents. should be handled and Sisposed of in accordance with the prevailing regulations land guidelines of agencies holding jurisdiction over the faboratery. REVISED: Aprit 1991 inTRo-sINTRODUCTION Lh Refer to the product label and to Materi 3 Standard Procedure for the Collection of Diagnostic Sheets for tals concorning: any seer Safety Data ‘00 Soecimens by Venipuncture. NCCLS, Sansara gets for details concerning any special precautions ‘elated to the handling of reagents, ASH-3," Vilarova, PA, Nationa! Commites for Chimes! Latoratoy Stancard (1977), “ iancard Procedures for me Collection of BIBLIOGRAPHY pea oa nacarein Giagnoste Blood Soecimens by Sun Puncture, NOCLS Kaeian LA and Pesce AG: Clinical Chemisvy ~ Teer, card x Stancard HA. Vilarova, PA, Natonal Commitee for ‘Aatiysis. anc Correlavon, St Lous, CV Mosby (198s) 93 (Cinta! Lanoratory Standard (1983) * Tote NW: Textbook of Cinica’ Chemisty, Phiadelon, PA, WB Saunders (1986) gp 478 - 518, TRO-6INTRODUCTION ‘Table 2 TECHNICON-DEFINED AND USER-DEFINED CHEMISTRY METHODS TECHNICON-DEFINED USER-DEFINED (SOFTWARE VERSIONS) CHEMISTRY Fa50] RA-1000| RAT | RAOGO—|RASOO|RA-TOO0 | RAXT [RA-2000 to | as | 2a | mar revease ‘METABOLITES ‘umn : : : : Calum : : : carbon cide : : . : (eneymate) Cholesterot : : : : Creatinine : : : + Cchionse : : : : ieee aati : : : + ” el cd Glucose : . : : HOL chotestot + . : : trorgantc oresprons | * . : : Magnesum : + : : Potasaum : + . . (coonmere) ‘Seam . : c ‘ (coonmete) Tota birwon : : : : : ” " Tota ron : : + : ota prcton : : : : Teayeendes (GPO) | * : + . ‘Urea ewrogen : : : . Ure ace + . . . ENZYMES AP : : - ALP (DEA) eof : aut zi : : : aut ae ee : fare Blok pt 5 Amylase : . . . ner : . . . ast : : : aste2 ele : cK . : : xe 2 : 5 cor . : : 5 4 o 5 : : : Loe-u ef fs : were elo te : ION-SELEGTIVE ELECTRODE (SE) CHEMISTRIES. Sodium (serum) s fe : ‘Sadivm (vere) : : : Potassium (serum) : : : Poxassum (une! : + : Carvon sone : : + NOTES: Usardetined metnd for ane conscted metoc, not ecammance te vee Behiomatic methods for aa and orc bkubiNs For use tnd Norn Amerca REVISED: April 1991 INTRO-7inTRO-8 INTRODUCTION Table 2 TECHNICON-DEFINED AND USER-DEFINED CHEMISTRY METHODS (Con) CHEmistRY TECHNICON-DEFINED (SOFTWARE VERSIONS) USER-DEFINED RAS00 RA-1000 28 RAXT | A200 23 | wat RELEASE RAEOO FA-1000 RAXT A200 THYROID. FUNCTION Thyroxine (Te)? ‘SPECIFIC PROTEINS Immuregiobotn 8 Immmunogiotuin G Immuconicbuin Transiern |. ca ca C-Reacwe Proten THERAPEUTIC ‘DRUG MONITORING Gentamicin $ Prencoarstal ® Phenyiin§ . ‘Theosnyioe & . Tooramycn § THERAPEUTIC ‘DRUG MONITORING APPLICATIONS Spa Emit? Ccaremazeone Syra Et vaproe Aco Syra emt Syra Em oincine Spa Emt Neacory procanamoe baucs oF ‘ABUSE Syaemedau Ae Syakmtdau 6* ‘Spa Emt Camano NOTES: “hyrone (Tlf 3 metnas using reformed reagents ‘TECHNION RastO00 Syste Gentamion and Totramyen ae rtomulated aud agents © On the TECHNICON fa-2000 ester Phencbar. Prenton. a ‘Se and Ems ave regsteredtacemarts a Sya Camgany, Po AO. CA. The Sa Em) Methods ae recent nthe slwaFe 35 2 anvenenca to customers. Rewsou Aoateatans Precets wil be senacie 23 fE8 te 4.24.13 wacomar of Swe Comaary alo At. CA . gs 8 “Consus ot ne lotonne cas tenzoiecoe meta matocuatneDrensyccra 4 DaDOMOrETE REVISED. April 1991, outage Non Amence for ine TECHNICON RA500 20INTRODUCTION ‘Table 3 AUTODILUTION ON THE TECHNICON RA-2000 SYSTEM eee NEAT RANGE HIGH VALIDATION RANGE HIGH {using Autoctuton) ‘Common Uae Common Units METABOLITES Avan 6a. wa Cau 1Smgct Na Caron sone ce a Chote 500 moe. 780 mol Crnatane 20 gc 50 mg. Chore 10 meat, 160 meat. rect anon ‘anes reg. a Ouenee 00 mor. 00 moi HOL cleo 135 moet a : \organe posses oma 20 no. Magtesun dome romp. on 5 meat Na poo 180 meat NA Tota bin ree romp oe. Toa von 1000 voit 2875 wot Tol pte 109 29a. Tyce (OPO) 500 moat 240 mol Urea nogen 100 moa. 200 roc Unease 25 0g NA eNzYMES ae 1500 Ue 23800 Ut ALP 1064) 2500 Ut NA ALT wo PSP 600 ut 2429 Un, aut Psp 20 UL NA Auta w PSP 50 Ut NA ‘amviase 200 Ut s200 Ut astwaPse 50 Ut 4000 UL AST 9 os asout Nm Astiaw Ps cou Nn ox 4f600UL 900 Un, cue 760 Ut NA cot hoot 500 Ut 0 200 ut 230 UL wry v500 UL NA wae nu 2000 UL Na on.setecrive ELECTAOOE (1SE) CHEMISTRIES Soda (som anc une) wa wa Potasoum (svn an se) NA wa ‘Caron once (tum oy Na NA THrr010 FUNCTION Tryroee 2 vont wa. | NA» Not Arpmabe AeWs¢0° Apa 1991 wwrro-INTRO-10 INTRODUCTION Table 3 AUTODILUTION ON THE TECHNICON AA-2000 SYSTEM (Cont) cuewsrny | NEATRANGEHGH — | VALIDATION RANGE HCH (utng Aan a eC | serarc anorens Pasian fone wa Srey 6 sao me ma omen ome wa Torn come mia teach Pn rwenapevtic onus mont Ofinc Cena eect 20 wa Carano oes sont = ree ee _ SPE aoe wa — ; : (quid reagents) 2 pom nas NIA Not Appicable REVISED: april 1991,MILES. Clinical Method Method No. SM4-0131E94 May 1994 System Applicability: TECHNICON RA® Systems ALBUMIN INTENDED USE This in vitro diagnostic method is intended to measure albumin concentration in human serum and plasma on a TECHNICON RA-500°, TECHNICON RA-1000°, TECHNICON RA-XT™, or TECHNICON RA-2000" system. Such measurements are used in the diagnosis and treatment of some chronic inflammatory diseases, collagen diseases, and liver and kidney disorders. For information regarding the use of these reagents on the TECHNICION ASSIST® clinical analyzer or the TECHNICON RA-100™ system, please consult the appropriate application guide, SUMMARY AND EXPLANATION The albumin (bromcresol green) method is based on the work of Doumas, Watson, and Biggs" who automated the original manual method of Rodkey.2 In 1965, Rodkey introduced bromeresol green (BCG) as @ specific dye for albumin. At pH 7.0, BCG dye Combines with albumin to form a stable complex, The ‘manual method of Rockey is an inverse colorimetric procedure that determines albumin as a function of the decrease in tree bromcresol green absorbance, Doumas ef af, automated the manual method of Rodkey. The modified method was converted to a direct colorimetric procedure by determining albumin asa function of albumin-BCG complex. Doumas et al found that bilirubin, up to a level of 80 mg/dL, and sodium salicylate, up to 60 mgidL, did not interfere with the method, Several additional reagent modifications serve to ‘optimize method response by minimizing the differences between the characteristics by which BCG dye binds to human and bovine serum albumins PRINCIPLES OF THE PROCEDURE ‘The serum sample is added to the BCG reagent to form the albumin-BCG complex The analytical solution is mixed for approximately 30 seconds, and the absorbance is measured at 600 nm. Abso‘bance data are converted into concentration unit, REAGENTS: ‘The following products are available in the package sizes listed in Tables 1 and 2: ‘Table 1 REAGENT PACKAGING ALBUMIN [ALBUMIN REAGENT REAGENTS ‘PRO Ne. OT int “For in Vitro Diagnostic Usa” Description ‘ALBUMIN REAGENT When formulated, one liter contains: Bromcresol {green 0.30 mmol, Preservative; Surfactant, Buffer {CAUTION Avoid contact with eyes. ski, for clothing Wash thoroughly” after Ranaing. Avoid ingestion. Reagent Preparation The Albumin Reagent is ready-to-use as supplied and ‘no preparation is required IMPORTANT! 00 NOT ADD WETTING [AGENT TO THIS REAGENT, ‘SERUM BLANK SOLUTION When formulated, one liter contains: Sodium Chloride 154 mmol, Preservatives, ‘CAUTION! Avoid contact with eyes, skin, ‘Wash thoroughly. after Randling. Avoid lageston, ‘Table 2 REAGENT PACKAGING SERUM BLANK pect ‘SOLUTION eee Roo No. RIOT Fer ue outde Noch Aenea STORAGE AND STABILITY ‘Te Albumin Reagent and the Serum Blank Solution storage and stabiities are showa in Table 3. Table 3 STORAGE ANO STABILITY STORAGE ‘saat vee [sn a ie Sexe cape om the IMPORTANT: 00 NOT STORE THIS REAGENT BELOW 10 *C ‘Serum Blank Serum specimens that are significantly hemolyzed, icteric, or turbid should be run with a serum blank to correct for any contribution of nonreactive components that absorb at the primary wavelength. The Working Serum Blank Solution 's used for this purpose, WETTING AGENT W (Prod. No, 721-1298-52) Contains: Surtactant CAUTION: Avold contact with eyes. skin, Sr clothing Wath tmoroughiy”atter Ranaling. Avoid vageation. — mites ZA Mies ine, Diagnastes Division “Tamown, NY 10581-5097 USA apaWorking Serum Blank Solution Preparation Add 3 crops of Wetting Agent W for each 20 mL of Serum Blank Solution, then swirl gently to mix. Label “Writing Serum Blank Solution.” When stored at room temperature, Working Serum Blank Solution is stable until the expiration date printed on the label of the ‘Serum Blank Sotution, SAMPLE HANDLING This method nas demonstrated serum’piasma ecurvalnce using itivm-neparn Soed collecion toes for pisma, Serum and plasma samples may be stored for one week at 2-"C to B°C or for one month at 30 “G. Frosen Samples should bo thawed at room temperature. ana mixed thoroughly before use. Thawed sampies Snous not be refrozen. ° Samples with albumin concentrations greater than 60 G1. 60 GH) should bo Sted mith isSone sate (8 Se Nach and reassayed PROCEDURE ‘The procedures for pertorming this method are located in the Operation Section of the Reterence or Operation ‘Manual forthe applicable TECHNICON RA system. Assay Parameters The assay parameters for this method are reprogrammed into each TECHNICON RA. system For additional details conceming these parameters, refer to the applicable Chemistry Program Date Sheet supplied with this methos. QUALITY CONTROL, It is recommended that the system be controlled using TECHNICON TESTpoint™ Assayed Chemistry Controls 1 and 2 (T03-1220-62 and T03-1221-62, respecively) ‘These products are intended to be integrated into’ Clinical laboratory's own quality control program ang procedures. These controls should be-assayed: 1. Atthe beginning of each eight-hour shit, 2. Whenever a new reagent (same or different lot) 1s used, 3. Following the performance of any detection system ‘maintenance or cleaning, A satisfactory level of performance is achieved when the analyte values obtained for each contiol are within the “Acceptable Control Range” published in the Package Insert provided with the TECHNICON TESTpoint ‘Assayed Chemistry Controls 1 and 2 NOTE: For consistent quality convo! resus, reconstute the contol products) as dracted ithe package ser) CALIBRATION The procedure to calibrate this method using TECHNICON SETpoint™ Calibrator, Prog. No. TOS 1291-62 (for use in North America) oF TECHNICON SETpoint Calibrator, Prod. No. TO3-1231- D0 (Hor use outside North America) is described in the aragragh tilled “Calibration of First-order late and Endpoint Tests” in the Reference or Operation Manual for the applicable TECHNICON RA system. ALBUMIN. TTECHNIGON METHOD No, SMé-o131E94 Reter to the package insert supplied with the calibrator for the system specific value (SSV) and handling instructions for the TECHNICON RA systems. RESULTS Albumin results greater than 6.0 g/dL. (60 g/L) on the TECHNICON RA-500 and the TECHNICON RA-1000 systems will produce numeric values and a “G" flag (On the TECHNICON RA-XT system, numeric values will be printed along with the message: “EXCEEDS HIGH RNG.” It the Endpoint Limit is exceeded on the TECHNICON RA-500 and the TECHNICON RA-1000 systoms , “YOOCOO™ willbe printed instead of the result along with an *L* flag. On the TECHNICON RA-XT system, “X0000K" will be printed instead of the result along with the message: “EXCEEDS EP. LIM." On the TECHNICON RA-2000 system, results are Broduced by the instrument for transmission to the various output devices. These devices include the 40-Column Instrument Printer and the TECHNICON RA-2000 system Workstation, which is an integral part of the system. if the data ‘used in the Production of a result have failed any part of the criteria for the production of a valid result. a flag is appended to the result upon output. Additionally. if the criteria are such that no meaningful resuit could be produced, the result itself is omited. Flags are converted to various text messages on the 40-Column Instrument Printer and these flags are presented in the flag field of the Workstation as a singie character. The following are ‘examples of what can be observed at the various output devices: Workstation Record: 1 the Range High for the albumin method is exceeded *G" will appear along with the result on the Workstation monitor and painted report. it te Endpoint Limit 1s ‘exceeded, “L” will appear in place of the numencal result on both the Workstation monitor and report 40-Column Instrument Printer: ‘Albumin resus greater than 6.0 gidL (60 gL) on the TECHNICON RA-2000 system, will produce numeric values along with the message “EXCEEDS HIGH FING” “if the Endpoint Limit is exceeded on the TECHNICON RA-2000 system, “XXXXXX" will be Printed instead of the result along with the message: “EXCEEDS EP. LIM.” The final disposition of a patient result on the TECHNICON RA-2000 system us handled through the system Workstation. If requested, @ patient result can be rerun via the Review and Edit “window of the Workstation. LIMITATION OF THE PROCEDURE ‘A number of substances have been reported to cause physiological changes in serum albumin concentrations 35 A comprehensive discussion concerning these and other interfering substances. their ‘Serum concentrations, and their possible physiological invoivements is beyone the scope of this publication au REVISED: May 1996, C aN‘As with any chemical reaction, users should be alert to the possible effect on results cue to unknown interferences fom medications or endogenous Substances. All patient results should be evaluated ia light of the total clinical status of the patient. Refer to Interpretation of Results contained in the INTRODUCTION to the Reference or Operation Manual for the applicable TECHNICON RA system EXPECTED VALUES ‘Tho range of expected values for the TECHNICON RA system albumin method is 2.5 gidL (35 gil) to 50 gat. (60 g/t. IMPORTANT. Ths range is prowded for eerence oly sen voraton should aetemana the lage oer values acteate tots ponuaton PERFORMANCE CHARACTERISTICS Performance characteristics of this method are essentially equivalent on all TECHNICON FA. systoms (TECHNICON. RA-500, TECHNICON RA-1000 TECHNICON RA-XT. and TECHNICON RA-2000 system). The following data applies to any TECHNICON RA system operating at 30 *C or at 37 *C, except for the TECHNICON RA-2000 system which has been validated at37 °C only imprecision Estimates of imprecision shown in Tables 4a and 45 wore obtained from replicate assays of a spiked human ‘Serum pools over a period of five (5) days. ‘Table 4a IMPRECISION (Common Units) tever | Tora | tora | worn | wren | ages | "ORM [teu] onan [waren coe | os | EP | ARS ve] ons | as] ons] as Tie 42 WeREEION (Un) wen | tore [ror | ween | acim] es | Tag | eae | ares | ree & |S | MBP | MS w [es [as] oe) as @ fim fas | te | 2 oiim 1 ie | 3 ALSUMIN TECHNICON METHOD No, SMs-oratESe Correlation Data The correlation data for this method are presented in Table 5, ‘Table 5 CORRELATION DATA i eer Sean ave | Sia ge | ML eau «| ea | “Behe natoo os L eSqeor > sci 22a] oser-o0]oaea] ox | 05-50 Sin or jswa? a 143 |0.966« + 0.1/0.940] 0.2 15-58 Sreon ese |] aH OT BOTY ESS ae x= Comparative sytem Y= TECHNICON RA system Linearity This method is linear from 0.0 gia (0.0 g/t) to 6.0 gid (60 gt). Sensitivity ‘The average sensitivity for this methoa is 0.048 A per Gia. or (0.005 & per gt) BIBLIOGRAPHY 1 Dounas BT, Waton WA, and Biggs HG! Aumn sandarss te measreren of sarum an win seeeaa grace Sin Chim Acts ana7-86 37H} ik Direct ssecropholometne determinaton of 2° Shure Fars Sth Cle Cho Vi rosy Tose) 2. Young 05, fect of Drugs on Cimes) Laboratory Tests. 3rd fs Young 0S 18 AACE Wasnegen OE (580) 4 Maren EW Hazard of Maceaton (Atrancer SP, Farage Ou ard Hassan’ WE i ogs) Pmadeiohar PA ang Toon Seas 8. Un aman ari ek 5. Constant NV ang Kabat HF. Onug-ndvoad moiscatons of Sburaiory tox vues = tenses 1605" Art J Hosp Beart ay REVISED: May 1994 au* CHEMISTRY PROGRAM DATA SHEET (TECHNICON RA-500* and RA-1000° Systems) ALBUMIN TECHNICON METHOD No, SM4O1S1E94 EY INTRODUCTION ‘The Chemisiry Parameters required to perform the albumin method are preprogrammed into the applicable TECHNICON RA-500" and the TECHNICON RA-1000" systems as a Technicon-defined method. CHEMISTRY PARAMETERS The parameters that have been preprogrammed are those that aro listed in the Common Units column of Table Ar. i the user wishes to report results in Common units, no parameter changes are required However. i results are to be reported in SI Units, the ‘Operator ‘must enter 8 and edit’ the preprogrammed parameters to coincide with those ised in the SI Units cokmn of Table A-t. Only those parameters that are shaded in the SI Unit columa are 30_| 0<5>30 Ret VOL 75.375 ut) | 7575 pu) UNITS Teo) | sen UNIT FAC 71.0000 | 10.0000 ‘DEGMAL 1 ao ROL LOW 2.000 0.000 RL HIGH (0370 0.370 RANGELO 00 ‘oe RANGE HI 60) 6 CALFACT, Seenotee | See notes SIDVAL Seenotef, | Seenotet NORMAL, 359 359 NORMAL H 503 508 SLOPE 71000 7.000 INTERCHT 0.0000 2.0000) PUN oozes | 0.0286 Eas ‘he number of chemisy parameters that con be accessed Sod eee arn eee ee Sauer tease ace oee carr ee sees resonate ne nmr eat es eeesam ae nea reeceraees ExiSpre crite mee oes eon Saag poh racateee ecera as os Bn Rte arcs oat Press to obtee 8 reliable mean CAL PACT value foes eee So cee TON Bn 2% enn PGs r sone: ee Teyetaer oS coe SS REVISED: May 1994 Asa)CHEMISTRY PROGRAM DATA SHEET (TECHNICON RA-XT~ Systems) ALBUMIN ‘TECHNICON METHOD No, SM4-0131E94 INTRODUCTION The Chemistry Parameters required to perform the albumin method are preprogrammed. into. the TECHNICON RA-XT™ system as a Technicor-defined method. CHEMISTRY PARAMETERS The parameters that have been preprogrammed are those that are listed in the Comman Units column of Table 4-2. if the user wishes to report results in common units, no parameter changes are required However, if results are to be reported in SI Units, the Operator must access the EDIT CHEM screen and edit the preprogrammed parameters fo coincide with those listed in the SI Units column of Table A:2. Only those Parameters that are shaded in the SI Unit column are different and require change. IMPORTANT: When 2 printout of parameters is ‘equese. only thom parameters ited bafece "yee wi appear on te rout Table A-2_ CHEMISTRY PARAMETERS ‘COMMON 3 PROMPT ‘UNIT unt enty | entay AME Teas | ns) aus Lock Seenoteb | See noted, ‘CHEMISTRY TyPE__| ENDPOINT | ENDPOINT SAMPLE VOLUME | 5@.5n) | S25m WAVELENGTH. 600 600 DELAY TIME 0:30 0:30) BLANKTYPE: BLANK | BLANE SEREAGENT VOLUME | _75(975 ul) | 75G75 mu) UNITS. 11600) | 6(ca) UNIT FACTOR 70.0000 DEC. POINT. oe BL LOW 2.000 ROL HIGH 0370 0.370 RANGE LOW ‘00 os RANGE HIGH 6.0 2 ‘CAL. FACTOR Senate | Seenotes STANDARD VALUE | See note? | Seenotef, NORMAL LOW. 350 383 NORMAL HIGH 509 509 SLOPE 1.000 1.000 INTERCEPT, 0.0000 0.0000. ENDPOINT UM 0.0786 0.0286 NoTes For compete aetais concerning the implementation of te trun refer othe paragon tied “Unit PaGtOn Sherry Botameter ia the eterence Manual fr the TECINICON RE Sesto [il Techrcondetined metnace contain 9 tok code which Hse ihe rambo enemy pronase con be acesnd on Charpes by re user ‘is ena can use tho ootona! sample barking pocedre. The ston of the bank soutien chon wen malang te reagent fay coniqsanon "A samele bla s chosen us the “SMR (CORA™ nbturtin of 2 worst. Although Ds values se exressd in common uns, the acta! “alves used wit the software se nternaay converted 0 5 uns neve soe ay ue Enter a inal val of | However belore patient sample are tly run, a ten CALBAATION FACTOR tale datermined wa ‘Binge spyoonsie comma Sa repeats hu be ued btm a reabe meen CAUSATION FACTOR value Ener ine aborgnate TECHNICON RA system SOV calbraton ‘hue (Conmon uns of unt B8apotcabie af ped nthe TTECHNICON SeTpotnt” carol aabage meet These cones ae prove tor reterence ony Ach aboratoy ‘heat Severe te Tange of Specie values 2plcna T0 Ee opueion = Lp(a.) REVISED: May 1994 ©*~ CHEMISTRY PROGRAM DATA SHEET (TECHNICON RA-2000- Systems) ALBUMIN TECHNICON METHOD No. SMAO1S1E94 SS INTRODUCTION ‘The Chemistry Parameters required to perform the albumin method ace preprogrammed. into. the TECHNICON RA-2000™ system as a Technicon-defined method. CHEMISTRY PARAMETERS The parameters that have been preprogrammed are those that are listed in the Common Units column of Table A-3. If the user wishes to report results in common units, 0 parameter changes are required However, if results are to be reported in SI Units, the ‘operator must access the EDIT CHEM screen and edit the preprogrammed parameters to coincide with those listed in the SI Units column of Table A-3. Only those Parameters that are shaded in the SI Unit column are 30 | 30 osss| ‘20°C we oP 1 85 | voex+30 | 30 loser| 5-210 ‘Gomosaive system or etnod 19> TECHNICON Ha system ug ceclera reagents) Linearity ‘When un without PSP actwvation, the method is linear to 600 UiL. This method. when run with PSP activation, 6 linear to 250. Autodiiuton is available as an ootion for the ALT metnod without PSP. and has been validated: to 3438 UIL at 37 "C on the TECHNICON RaZ000 system only. Refer to the INTRODUCTION section of the Methods Manual, which describes the autodilution ‘option. 1 Without PSP ecovaan ony ALT WAWO 950-4 Al TECHNICON METHOD No. Swe-orsaesa Sensitivity The sensitivity of the ALT method with or without PSP activation is 0.0002 absorbance (A) unit per minute per ‘unit of activity (UL) at 37 °C. BIBLIOGRAPHY Wroblewsk F and Laue J: Serum glutame oynuwic in caraae, and nesase disease, Brot Sos Exper Bil and Wed 81°385 (1956) 2 Bergmeyer HU. Schebe P, and Wanlelela AW: Optmzaton of mettous ior asoarate ‘amnoiransierase and alanine amnovansierase. Gin Cnem 2458-75 (1878) 3 lewraicnas Federavon of Canal Chemistry. Commtee on Standaras, Enzyme. Pane! (Saris NE ed): Provisenal fecommenaavons on FEC motnads tor the measure of GBRNRE concencatons oF enzymes. ‘Cin Gram Se T20-721 1970) «Batra S200 Most OW: Aocenryme of aspanae ‘Mmowransterase in serum in Nealin and daease. in Cham acta T3263 (97 5 Hatkenscned JCM and Oyt CCM: Oetermnaton of serum arproransieases: Aciwaton oy pytogsal's:-ohosanate ‘elaton w suoswate concenwaton, “im Chem 211758 (sre © Tetz NW: Textbook of Cimical Chemistry. WB Saucers Co. Prisco, PA (1966) p O76 7 Young 0S. Etfect of Orugs on Cimea! Laboratory Tests. ars fe: Young OS ea. AACG, Washington DC (1930) 8 Maron EW. Hazara of Meaicanen (Alesanger SF. Farage Ov. 30d Hassan WE Jr. eas) Pradetoha, PA” ang Teron. Canada. 1/8. Uopiicon Company (1971) 9p 169.189 8 Constanino NV ang Kabat HF: Drug.induced moaicatons of \aboratery test values = remsod 1959, Am J Hose Brana sozer 1979), REVISED: Mav 1994CHEMISTRY PROGRAM DATA SHEET (TECHNICON RA-500° and RA-1000* Systems) ALT W/O PSP TECHNICON METHOD No. SM4.0104894 SN INTRODUCTION ‘The Chemistry Parameters required to perform the ALT method without PSP are preprogrammed into the applicable system as a Technicon-defined method beginning with versions 1.9 software (TECHNICON RA-500° system) and 2.8 soltware (TEGHNICON RA-1000* system). CHEMISTRY PARAMETERS The parameters that have been preprogrammed are those that are listed in Table A-1 ‘Table At CHEMISTRY PARAMETERS COMMON Load siuntrs ENTRY CREM» aur Loc. Seenotes Te @IZERO ORDER TNVERSE CO vEST % swe voL 3eC18 uo, FALTER ewe 340) ‘BCFACT 7.0000. ‘DEP uM. 0.2800 DELAY. T30 enGT VoL 166 (330 uw) ‘UNITS 3 (uA) NIT AC 0000) DECIMAL, 0 Low 2.600) RANGE LO ° RANGE WI 600. TCALFACT. Seeroteb AGT RATE Seenotee NORMAL se NORMAL mo SLOPE. 1.000 INTERCT 2.00007 Cht0e-6 0.00 2108-6 $3999.07 Dr.1086 20.00) DELTA —t 2.008, REVISED: May 1996 NOTES AN Tecnacondetined mathods conan 4 cx cade whch inde hanged oy the user Technean-elned rrcaraer lets are orgie at he tector, {ind 00 ot requreroutna on-staeasiraton Pater (9 te S004 ‘Samos spews be used Wo odio 9 enable mean AGT RATE ee ‘These ranges 2f@ toned for rterenca only Eien laboratory ‘Pou determine the Yarge ot emecied wes Sopcabe 1H soosaton ALT Wrwo PSP (A-1)CHEMISTRY PROGRAM DATA SHEET (TECHNICON RA-500° and RA-1000* Systems) ALT W PSP. TTECHNICON METHOD No, Sie-o134894 —_————————— INTRODUCTION ‘The Chemisty Parameters required to perorm the ALT method with pyridoxal S-phosphate (PSP) are reprogrammed into the applicable system as a Technicon-defined method beginning with versions 1.3 software (TECHNICON RA-800° system) and 2.8 software (TECHNICON RA-1000° system). CHEMISTRY PARAMETERS The parameters that nave been preprogrammed are those that are listed in Tabla A-2. IMPORTANT: whan a printout of oarameters ss ‘quid. On ose parameters Qented alee ‘ype ml pce! onthe peru ‘Table A.2 CHEMISTRY PARAMETERS common | baotd siunrrs tra came TAP Loor Seenotes Tye CZs OnE TAVERSE ‘ayes sup vor 36080) FALTER wa 3407 merACT 10000 Oerun 0.3600 DELAY S00 act vot 6.0300) UNITS aut) Uumreac 0000 peoMaL ° RaLLOW 758 RANGEL ° ANGE HI 350 CALEACT Tee notes AGTRATE Seenotec WORMALL $2 NORMALH a0 SLOPE 7.005 TATERCPT 2.0000 1.0 2.08 a.t0es 3299807 DiA0ES 1200 DELTAr e008 ALT WIWO P5P(A-2) NoTEs AM Techncon-detne metnats conan 3 ck sade whch Sis ‘he uber of chery parameters tht can ow moms ‘hanged Oy th ure and do ot reaure rota arta caravan Reta othe sacle ‘Wied “CALRATION” ct the appsabie TECHNICON Ra system Retarence Manual ecairaten of tr-arae tess 9 reaared A-RGT RATE wave @ onermned by a” RB rato assay aS escnoed in The Relevance anual Tor the sooicable FECHNICON RA’systom The RE ate autby ve eauedied wn the aporoprate uncammane code” Su Teohcelas a water Ssumows shoud be used to etiam 2 roan mean AGT RATE ‘These ranges are crowed tor releence ony Each abort shoud determine ine Yonge o! eroccioa vives aoptcta 10 ts onstaten REVISED: May 1994CHEMISTRY PROGRAM DATA SHEET (TECHNICON RA-XT> Systems) ALT.WIO PSP TTECHNICON METHOO No. SM40134E94 INTRODUCTION ‘The Chemistry Parameters required to perform the ALT method without PSP are preprogrammed into the TECHNICON RA-XT™ systom as a Technicon-detined ‘method beginning with version 2.3 sottware, CHEMISTRY PARAMETERS ‘The parameters that have been preprogrammed are those listed in Table 8-1 IMPORTANT: whan a orntout of parameters 1s quate. ony these pataraters Sted an Baokace ‘Table 8-1 CHEMISTRY PARAMETERS common rnomet siunrrs ENTRY. aM Waar LOCK Seenote a, __ tr ZERO-ORDER INVERSE CHEM VES SE SAMPLE VOLUME 360800) WAVELENGTH 340 BICHROMATIC FACTOR 7.0000) DEPLETION UMIT. 0.2800 DELAY TIME 1:30 SE REAGENT VOLUME 603050) UNITS: ut [UNmT FACTOR 2000) DEC. POMT 2 aL LOW 600) RANGELOW, 2 RANGE HIGH 00 ‘CAL. FACTOR See nore, REAGENTRATE See note (NORMALLOW 5 NORMAL HIGH a0 SLOPE 7.000 INTERCEPT, 0.00001 C1.106-6 0.00 C1086 3399903 DI-106-6 20.00 DELTA #. 0.008 REVISED: Mav 1994 ‘reranet meant we peters ae cen Nimes Pata arte at Sisco sere nome te aes ig oe SE Ba Sutcommara\ code "Su rohan ot uate spies Sone $e seviioeTab clea tines ‘enna Soe een ot ae SSS SS Se, Se amsCHEMISTRY PROGRAM DATA SHEET ALTWPS5P . (TECHNICON RA-XT~ Systems) TECHNICON METHOD No. $¥4-0134E94 INTRODUCTION, Tho Chamty Parameters equre to pert the ALT mmathea wih pyncocal ovosaneer trey ste moprgannad io he TECInNceR Bade. ese S52 Tecmcon-detned matos tegenrs oan ee Ea totwan CHEMISTRY PARAMETERS The pavanotrs tat have boon preprogrammed are tse iso Tate 9 uroRtant, wren + omiaut o seamen meen Sabin ol oven sms Table 8-2 iewisTRY eaRAnETERS noes +A tenement mT San uke nt ‘COMMON {the number of chemesty parameters that can be accessed and e meee PROMPT. ae 1D Technicon-aetined zero-orcer ts ara oecatyateg ote can ar ROSES Tareas a ae Meee racial geomet ney ioe Seana © AGEN a aur oy ee ett ma Taboes BERS Seta teenies te ermeea INVERSE CHEM YES, ‘Sees corn enaie meer REGENT METS eee? hae Teneo —}—a ¢ Sipe Sesto on ey Sanaa a Baie es eS Se, a TOUGHT HRSTOR = CERN EAT sear € DELAY TIME 5:00 — Sacer toma —| —wees om i ie tae Siero a Sar aa Tate n anata = ares on ae a é fiom ae a tare a aT — aa im eet ann aie ma mae aa ALT ww PsPre CHEMISTRY PROGRAM DATA SHEET (TECHNICON RA-2000- Systems) INTRODUCTION ‘The Chemistry Parameters required to perform the ALT method are preprogrammed into the TECHNICON RA-2000™ systom as a Technicon-detined method. CHEMISTRY PARAMETERS The parameters that have been preprogrammed are those isted in Table C-1 IMPORTANT: When 2. prntout of parameters 1s ‘mgumnd ony tose parameters grad wr baurace ‘Table C-1_ CHEMISTRY PARAMETERS. common Promer sites cn mae Gaar Lock See notes Tyee ZERo-onDER TaVERse CHEN Yes Si samnis vouunie Bees TALE saris VOU Cor WAVELENGTH 7 BEGROMATICFACTOR 7.0000 OEPLETION Liat 23400 DELAY Tie 30 TREAGENT vou ows uns rm Dhar racroR 008 OF<. PONT 3 aaLLoW a8 RaNceLow ° RANGE GH 0 Wal RANGE HIG aa GaumRaTiOn AcTOR See rane MEAGENT Rae See note NORMALLOW HORMAL i SLOPE iarencerr eit08 0 e084 Tana Bi-tob 200 oatay ace REVISED: Mav 1994 ALT W/O PSP TECHNICON METHOD No, SM4-o134696 NOTES A Techrcanetinad mathods conta lock code whch fits tne numberof chersiy parameters tat an be aueasse So ances bythe wee ‘Teenncon-etned coo-rcer tesa orecabrsed atthe factory {F560 no reaure four on-atecasrapan Pelt 1 te section hed “CALIBRATION ot ine TECHMICON RAc2000 System) Goeravon Manual recteraton of tarde ats rea ALREAGENT RATE value catermned by an RG ate asoy 25 SRE un EMCO AR a Ort ‘Bomar case "Sa easates ot tee rows be {S500 cota a voanie moon REAGENT Aare ‘These ranges are ered or raerence ony, Each laboratory nou Setermie the range oI epesied ous sopmcaote 10 ALTWwrwo psBce-1)—————————— dl HEMISTRY PROGRAM DATA SHEET (TECHNICON RA-2000° Systems) ALT W PSP TECHNICON METHOD No, SM4oTS4E94 —e_e———— INTRODUCTION ‘The Chemistry Parameters required to perform the ALT method with pyridoxal S-ohospnate (PSP) are reprogrammed into the TECHNICON RA-2000" system as a Technicon-defined method. CHEMISTRY PARAMETERS ‘The parameters that have been preprogrammed are those listed in Table C-2, IMPORTANT: when s oontout of oarameters ss ‘queso. onty ese sarametrs oredr basic ‘joe mt eopme’ on te pan Table C-2 CHEMISTRY PARAMETERS conan rower sins in Tae (aaa toe 1 ses nates Tre Teno. one TaVERSE Git Tes S samrce vou Ba WAVELENGTH 30 TGROMATICENCTOR 000 OEMETION LAT 0.3600 OeLAY Tae $20 Stacent vounie wax ‘uM ue Dar Facron 000 ‘BIC PONT 7 ABLLOW a Rance Low ° AaNGE WG 358 GHuusRaTiON FACTOR | Teerate REAGENTRATE See notes NORMAL LOW se HORMAL wg | sp sLoPe [08 warencerT | poo eiet0e 00 Gatots mnt 1.108 1200 OuTAs 008 [ALT WO PSPC.2), REVISED: May 1996 NoTEs [AN Tecancon-cetines matrens conan 3 lock cade wen Mes {mq ruber of chemay parameters tat can ow memes on ‘hanged by tha toa A REAGENT RATE vane a gelermong by an RB rate tsay 35 ‘eer mine TECTONCGW BS t000 Sysiom Conran aru Fhe'RS fate" assays cn reauesieg ‘Cases oe ‘eg to cotain a veuaoie moan REAGENT Bate woee These tances are oronoes or reterance ory Each aberstonMILES Clinical Method Method No. SM4-0157E94 May 1994 ‘System Applicability: TECHNICON RA® Systems ALANINE AMINOTRANSFERASE (ALT/P2), EC 2.6.1.2 (Two-reagent Method, With Pyridoxal 5-Phosphate [P5P]) INTENDED USE ‘This in wiro diagnostic method is intended to measure alanine aminotransferase activity in human serum on a TECHNICON RA-500*, TECHNICON RA-1000°, TECHNICON RA-KT™, or TECHNICON RA-2000™ system. Such measurements are used in the diagnosis ‘and treatment of liver disease and in monitoring the course of veament for hepatis and acive posmecrote This diagnostic method is not intended for use on the TECHNICON RA-100® system. SUMMARY AND EXPLANATION ‘The ALTIP2-method is based on work by Wroblewski and LaDue,+ who, in 1956, originated a procedure that Coupled Lactate Dehydrogenase (LD) and NADH to the aminotransferase reaction. This procedure has been modified by H.U. Bergmeyer to eliminate side reactons ‘and to optimize substrate conditions.? Subsequently, the Intemational Federation of Clinical Chemisty (FCC) defined conditions for the reference method. The ALT reagents are formulated in accordance with these IFCC recommendations. PRINCIPLES OF THE PROCEDURE ‘The reaction is inated by the adcition of e-ketoglutarate ‘as a second reagent Alario ar rte ectetogiurarate cahiamate Pynevate + NADH —!2 > Lactate + NAD The rate of decrease in the concentration of NADH is rectly proportional to the ALT activity in the sample. ‘Absorbance data are converted into reportable activity values (UL) based on calculations for zero-order kenetics. Pyridoxal S-phosphate is the coenzyme in the alanine aminotransierase reaction. ‘ts presence in the reagent activates any available unsaturated apoenzyme (enzyme without coenzyme) and increases the measured ALT mes REAGENTS: The following products are available in the package sizes fisted in Table 1 Table 1 REAGENT PACKAGING. ‘Table 1_REAGENT PACKAGING (Com Se ear ee SS LSS moe | mute | nuts a esc aPatloe 1 When reconstituted, one Miter contains @ 37 *C: ‘a minimum of Lactate Dehydrogenase (porcine ‘ugcle) 2500 U; and L-Alanine 600 mmol, NADH 0.28 mmot Sutter; Stabilizer UNE Acid contac win eps, hn ‘lothing. Wath thoroughly after ncting- Avoid ingestion. ALT REAGENT 2 When reconstituted, one liter contains: a-Ketogiutarate disodium 180 mmol CAUTIONS Avoid contact with ayes, skin, ‘or clothing Wash thoroughy ater Ranging. Avoid ingestion PYRIDOXAL PHOSPHATE (PSP) REAGENT ‘Contains: Pyridaxal §-phosphate ‘CAUTIONE Avoid contact with eyes, skin, ‘coming. Wash thoroughly” after Rrcting- Avoid ingestion WETTING AGENT W (Prod. No. T21-1298-52) Contains: Surtactant CAUTION Avoid contac. with ayes, akin. or coming Wean moroughiy after Randing. Ared ingestion. Reagent Preparation ‘The ALTP-2 reagents preparation instructions are specified in Table 2. ‘Table 2 PREPARATION INSTRUCTIONS = worn ‘WORKING. ALP REAGENT 1 ALT REAGENT 2 THi7e1 | 10 mL" | 2drops | Tete | 20 me THii7e | 20 m0" | Sarops | Thies | come {Um ow (PP eet FF Use wo [2 PP abe Working ALT Reagent 1 with PSP ‘To prepare the 10-ml. size, reconstitute ALT Reagent 1 by pipetting exactly 10 mi of distled water. Add t tablet of Pyridoxal Sphosphate, and swirl gently until the PSP is completely dissolved. Add 2 drops of Wetting ‘Agent W, and swift gently to mix. Label “Werking ALT Reagent ? with PSP." Abbreviations: NAD (ncotnamide acenme dinucieoncel: NADH (eeauees NAD) mnt PARAMETER Poo NO. soar TRE ROO. NO. srsre: sirrae arcu geiome | exome Rar PACO. NO. issree ruses ‘wroute team ‘exert mites JA Miles Inc., Diagnostics Division TTarytown, NY 10591-8087 USA ALP-on- To prepare 20 mL, reconstitute ALT Reagent 1 by pipetting exactly 20 mL of distiled water. Add 2 tablets Of Pyridoxal S-Phosphate, and swirl gentiy until the PSP is completely dissolved. Add 3 crops of Wetting Agent W and swirl gently to mix. Label “Working ALT Reagent 1 with PSP.* Working ALT Reagent 2 To propare the S-mL size, reconstitute ALT Reagent 2 by pipeting exactly 3 mL of distiled water, Swirl gently tomix and label “Working ALT Reagent 2" -OR- To prepare the &-mL size, reconstitute ALT Reagent 2 by pipeting exactly 6 mL of disiled water, Swirl gently toimix, and label “Working ALT Reagent 2" STORAGE AND STABILITY ‘The ALTIP2 reagent storage and stabilties are shown in Table 3. Table 3 STORAGE AND STABILITY some | saa |e? te tea ay ote mas roar] wee (ene A Rear ad en P| [or snl trea ee ee fERSigeares ee [techntcon Reso meron ‘SAMPLE HANDLING Specimens are best stored retigerated. Minimal loss of enzyme activity occurs at 0 °C to 4 °C over 1 to 3 days® Frozen samples should be thawed at room temperature and mixed completely before analysis. ‘Thawed samples should not be reftazen. ‘Samples with resuits that exceed the linear range of this ‘method should be diluted with § gidL (60 g/l) human serum albumin or isotonic saline (0.9% NaCl) and reassayed. PROCEDURE ‘The procedures for performing this method are located in tho Operation Section of the Reference or Operation ‘Manual for the applicable TECHNICON RA system. Assay Parameters The assay parameters for this method are preprogrammed into each TECHNICON RA-500 and TECHNICON FA-1000 system, for use outside North America, and into each TEGHNICON RA-XT and TECHNICON RA-2000 system. For additonal details Concerning these parameters. refer to the applicable Chemistry Program Data Sheet supplied with this method. (aLtie2) ‘TECHNICON METHOD No: SM4O1S7ESS QUALITY CONTROL be controlled using TECHNICON TESTpaint™ Assayed Chemisty Contvols Tand 2 (T03-1220-62 and TOs1221-62, respectively). ‘These products aro intended to be integrated into a Glinieal‘aboratery's own quality contol program and procedures. ‘These controls should be assayed: 1." At the beginning of each eight-hour shift 2. Whenever 2 new reagent (same or diferent lt) is sod. 3, Fellowing the performance of any detection system ‘maintenance or cleaning. A satisfactory level of performance is achieved when the analyte values obtained for gach contol aze within the “Acceptable Control Flange” published in the Package Insert provided with, the TECHNICON TESTpoint ‘Assayed Chemistry Contols 1 and 2 NOTE: For consist qualty conto resus. (eonsine the contol proces) a rocted in he package insert). CALIBRATION ‘The instrument is precalbrated at the factory and does rot require routine on-site calibration. However, the procedure described in the paragraph titled “Calibration Of Zero-order Tests" (see Tab Section titled “CALIBRATION” in the Reference Manual for the applicable TECHNICON RA syste) can be used to verify method calibration or to recalibrate the method should either become necessary (see paragraph tied INTRODUCTION, in Tab Section, “CALIBRATION’). RESULTS For a detailed explanation conceming the calculation used 10 report the result for this zero-order chemist, refer to the TECHNICON RA systems Reference Manual ‘Section tiled “Calculation of Results.” ALT/P2 results greater than 650 U/L will be printed out with cumeric values by 2 7G" flag on the TECHNICON RA-500 and the TECHNICON RA-1000 systems, The results will be printed out with the fnumerie values followed by the message “EXCEEDS HIGH ANG" on the TECHNICON RA-XT system. Ifthe ‘Mean Square Error Limit (MSE) D1 or C2 (see the applicable TECHNICON RA system Reference Manual Table 7—#) TECHNICON RA System Reference Manual is exceeded for the TECHNICON RA-500 or the TECHNICON RA-1000 system, an *L” flag will be brinted with “Y00O00C instead ‘of the result, On the TECHNICON RA-XT system. ifthe MSE (D1 or C2) kit is exceeded, “Y000KX" will be printed instead of numeric results followed by the message “EXCEEDS MSE LIM". Ha result exceeds the Depletion Limit on the TECHNICON RA-500 or the TECHNICON RA-1000 system, “XXOOCK™ will be printed instead of the result Preceded by @“D" flag. On the TECHNICON RAXT System, numeric resus wil be replaced by “X00000" ‘and followed by the message “SUBSTRATE OEPLTO.- aun REVISED: May 1998On the TECHNICON RA-2000 system, results are Produced by the instrument for transmission to the various output devices. These devices include the 40-Column_ instrument Printer and the TECHNICON RA-2000 system Workstation, which is an imegral part of the system. if the data used in the production of a resut have failed any part of the criteria for the production 6 a valid resut, a flag is appended to ‘the result upon output. Additionally if the criteria are such that no meaningful result could be produced, the resuit itself is omited. Flags are converted to various. text messages on the 40-Columa instrument Printer and these flags are presented in the flag field of the Workstation as a single character. The following are rambles of what canbe cbsorved atthe various ouput ievices: Workstation Record: H the Range High for the ALT/P2 method is exceeded, 6" will appear along with the result on the Workstation ‘monitor and printed report. if the Substrate Depletion Limit is exceeded, “D" will appear in place of the numerical result on both the Workstation monitor_and fepor. if the Mean Square Error Limit (MSE) is ‘exceeded, *L" will appear in place of the numerical result on both the Workstation monitor and report. 40-Column instrument Printer: ALTIP2 results greater than 650 UML willbe printed with the numeric resut folowed by the message: “EXCEEDS HIGH RNG* on the TECHNICON RA-2000 system. ff the Mean Square Error Limit (MSE) D1 or C2 is exceeded, “20000X will be printed instead of the ‘umeric result along with the message: “EXCEEDS MSE LIM" if a sample causes the Depletion Limit to be ‘exceeded on the TECHNICON RA-2000 system, the ‘numeric result will be replaced by "Y0OOQKX"” and followed by the message: “SUBSTRATE DEPLTD." The final disposition of a patient result on the TECHNICON RA-2000 system is handled through the system Workstation. If requested, a sample can be rerun via the Review and Edit “window” of the workstation. LIMITATION OF THE PROCEDURE ‘A number of substances have been reported to cause Bhysiological changes in serum alanine aminotranierase concentrations.’* A comprehensive discussion conceming these and other interfering substances, their serum concentrations, and their possible physiological involvements is beyond the scope ofthis publication. As with any chemical reaction, users should be alert the possible effect on results due to unknown interferences from medications or endogenous substances. All patient results should be evaluated in light of the total cinical status of the patient. Refer to Interpretation of Results contained in th INTRODUCTION to the Reference Manual for the ‘applicable TECHNICON RA system, EXPECTED VALUES ‘The ranges of expected values for this method at 30 °C or 37 *C. except for the TECHNICON RA-2000 system which has been validated at 37 *C only. are shown in Table 4 REVISED: May 1996 (aLtp2) TTECHNICON METHOD No. SMA-O1S7EB4 ‘Table 4 EXPECTED VALUES TOWERUMAT | UPPERURAT mr un, wn 30 2 0 a 3 2 TMPORTANT: These ranges ve peed 1 rafererce ony. Exch ubosiay su caren te ange Of ‘Soocandvlos sppictie © popRSoN. PERFORMANCE CHARACTERISTICS Performance characteristics of this method are essentially equivalent on all TECHNICON RA systems (TECHNICON RA-500, TECHNICON RA-1000, TECHNICON RA-XT, and TECHNICON RA-2000 systems), ‘The following data apply to systems operating at 90 °C oF 37 *C, except for the TECHNICON RA-2000 system which has been validated at 37 °C oniy. imprecision The estimates of imorecisin, obtained win and without ap actvaton. from repcate assaye of spiked aman serum pools that were un over a period o ive () days ae shown Table Table 5 MPRECISION Tort | ware | ea yen | TSE | TE | NSS | Money ey | oo _| Mea? | Moss ma) 33 | es] as | as wo | sx | as | 42 | aa so_| mis | 20 | ae | 08 TROTE The wken n UL nated in Tabs Abad @ ae te sone nS wie Serra nt Correlation Data The accuracy of the ALT method using dry-biend reagents (y) was determined by correlation studies with the previous ALT methods which used lyophilized reagents (x). The regression fine parameters obizined from the corelation studies involving ALT with PSP ‘ctivaton at 37 °C and 30 °C are presented in Table 6. “Table § CORRELATION DATA mange Jcowpararve| | peansssion | g,. oe x | “Eouanen «| exer or w=) ow ‘CONC metmoo cone sueaieass [os | ae +30 | a0 [asa] «19 ‘Sucowsrasa | 68 | tare-28 | 30 asa] 5-210 re 3 Comparative system or metiod Y= TECHNICON RA Systm (ung dy bend agents) Linearity ‘This method is linear to 650 UA. at 37 °C. ‘Sensitivity The sensitivity of the ALT method with or without PSP. activation is 0.0002 absorcance (A) unit per minute per Unit of activity (UI), at 37 *C. aureaBIBLIOGRAPHY * Serene ‘near. b hepa eae Ee Sos Exper Biol and Med 91:589 (1956) Pros Se 2 Serpe HV, Senate P. and wand AW: Opsrion Yor Sepetano “sminoranserase and ale ‘Gin Chom 2050-73 (1579) 3 ot a Standards, Eneyme Panel (Sans NE ed): Provisional ‘or FCC menace 4 Ramake $ and Moss OW: Apoenzyme of asparate Smnovansierase in serum in healn and disease. Gin Gum det 7228" Ga (arte) TTECHNICON METHOD No. SM4-0157E54 tere os coer aoa © subse ‘Sncenrason lin Chem 251155 ‘ re eciel Oniy. WOSanies SEM a ee ep esos tT 28 ‘era Sa Lice ae as {8 Marin EW: Hazard of Mectaton (Alexander SF. Farage OM, fand Hassan WE dt) Priacoonia, PA and Toren, Sihada 18. Upaentt Company (1971) po 169-189 Constante NV and Kabat HF: Onig-induced mocticatons of (Ege ‘ex values = rewsed 1073. Am v Hosp Pharm sozert (era) auras FEVISED: May 1996Sed CHEMISTRY PROGRAM DATA SHEET (TECHNICON RA-500° and RA-1000° Systems) INTRODUCTION The Chemistry Parameters required to pertorm the ALT/P2 two-reagent mathod are preprogrammed into the applicable TECHNICON RA;S00° and TECHNICON RA-1000* systems 2s a Technicon-defined ‘method. ‘This method is applicable for use on systoms ‘outside North America. CHEMISTRY PARAMETERS. ‘The chemistry parameters required to perform this method on a TECHNICON RA system have been reprogrammed into the system. The parameters that have been preprogrammed are those that are listed in Table At. These parameters apply whether the results «ae to be reported in Common Units or in SI Units, since results are reported in Intemational Unitsfiter (U/L) in both unit systems. IMPORTANT: wnen a printout of parameters is ‘vest. ely Bese parts Sted bolace ‘hom wil seat onthe pn ‘Table A-1 CHEMISTRY PARAMETERS ALT/P2 (INTERNATIONAL) ‘TECHNICON METHOD No, S¥s-o157E94 co monet aus on = ae = aed Cc TPE. [C.ZEAO ORDER) ae a % SMP VOL T 0.20) are ee co I eee i ea a ie ae ea eer re ae se UNITS: 1 3 (ut) a = a : ae os as : aoa = ome ao a oe = a a = = a = IMPORTANT: To. pero ts method, enter a delay of 400 minuite in response (© the 2ND AGT DELAY proms wich apcears olowng te Sperate corsnand. REVISED: May 1994 wee Tacs ranges are rovted for reference any. Each laberatary pir ‘be roe aureziact)—— CHEMISTRY PROGRAM DATA SHEET ALTIP2 (TECHNICON RA-XT™ Systems) TTECHNICON METHCO No, SMe-a157E54 INTRODUCTION The Chemisty Parameters required to perform the ALT/P2 method are preprogrammed into the TECHNICON RA-XT™ system as a Technicon-defined method, CHEMISTRY PARAMETERS The chemistry parameters required to perform this method on a TECHNICON RA-XT system have been [preprogrammed into the system. The parameters that hhava been preprogrammed are those that are listed in Table A2. These parameters apply whether the resus are to be reported in Comman Units or in Si Units, results are reported in Intemational Unitsfitor (WL) in both unt systems. Hl IMPORTANT. Won + pint af armas i eae er oe Se Sa Selo le ss Table A-2 CHEMISTRY PARAMETERS 18 Tennant cain ck ca wh its in al lyon pp | ec es PROMPT aS, ‘emicatred wr as ae recalatesat facory penalty cpa Cua TCANGRATION™ of the TECHNICON RAXKT System Tie AE a ee ee freee mene cree aoe sa aa ey PE za one 22 eee VERSE ove = Soa 3% SAMPLE VOLUME. (60,30 pt) Bee eee xe ovat toy terre =e EC oe Ro a Se TemguATC HEMET ves TOMOMATCPACTOR om CERETON LMT te (tearm Taxa von Seow, SENDAI = =m aesceer vou cana ONT wt Ta TOR Ts TEC PONT 3 oe Tas TANGO . tance a aL recTor Teen MSE a ORAL Low = Aone a See a wre one eriaes can rors estat ores 3000 eta p oara IMPORTANT. To perm tis meted ener a cay of 00 ‘minutes in response tb he ZNO AGT DELAY promot whch ‘pears alow te operate command. ALTP.2(8.2) REVISED: May 1894 Cc——_ CHEMISTRY PROGRAM DATA SHEET (TECHNICON RA-2000™ Systems) ALTIP2 ‘TECHNICON METHOD No. SiMs-o157E94 INTRODUCTION ‘The Chemistry Parameters required to perform the ALT/P2 method ace preprogrammed into the TECHNICON RA-2000™ system as a Technicon-defined method. CHEMISTRY PARAMETERS ‘The chemistry parameters required to perform this ‘method on a TECHNICON RA-2000 system have been preprogrammed into the system. The parameters that have been preprogrammed are those that are fisted in Table A. These parameters apply whether the results are to be reported in common units oF in Si units, since: ‘esuits are reported in International Unitsfiter (U/L) in both unt systems. IMPORTANT: When a. seetout of parameters is ‘Table A‘d_ CHEMISTRY PARAMETERS. rious eer a a en oe ian ener ee rere = ei aoa aoe a Scemncoearey Ss eee coe mee eae pases Eos creer cara om one = ee os a me ie a A a i ieee ; eet a See a ee oe ieee iF eect a a me ea a aaa oes a IMPORTANT: To pecorm he meiod ener a dey o 00 ‘anuts in response © be 2ND AGT DELAY promot, REVISED: May 1894 A REAGENT RATE sae « 288 cae samy Seabee el ArZa00 Systom Oneraten aru She Ro gy ALTP2 (43)MILES. Clinical Method System Applicability: TECHNICON RA® Systems ‘Far in Vitro Dlagnostic Use” ALKALINE PHOSPHATASE (ALP) EC 3.1.3.1 (2-Amino-2-methyl-1-propanol Buffer) INTENDED USE This in vitro diagnostic method is intended to measure alkaline phosphatase activity in human serum and Plasma on a TECHNICON RA-S00" TECHNICON RA-1000°, or TECHNICON RA:XT™~. and TECHNICON RA-2000* system. Such measurements ‘are_used mainly in the dagnosis and treatment of hepatobiliary and bone disease. For information regarding the use of these reagents on the TECHNICION ASSIST® ciinical analyzer or the TECHNICON RA-100™ system, please consult the appropriate applications guide. ‘SUMMARY AND EXPLANATION The alkaline phosphatase method is based on the work of Tietz et ai.' Test conditions. as used for this method, hhave been proposed to the AACC Subcommittee on Enzymes as the standard method for measuring alkaline Phosphatase activity.? The proposed method is a Modification of the original Bessey, Lowry, and rock method" with p-sitropheny| phosphate (PNPP) PRINCIPLES OF THE PROCEDURE The reaction is initiated by the addition of patient sample to the reagent Pritrophenyl phosphate 42 6 prtophenol substrate, pn A 2-amino-2-methyl-1-propanol (AMP) butter is used to ‘maintain the reaction pH at 10.3 to 10.4. Magnesium and zine ions are added to the AMP butfer to activate ‘and stabilize the enzyme. During the reaction, the alkaline phosphatase hydrolyzes the PNPP substrate to form p-nitrophenol which is colored (yellow) and provides its own chromogen. The reaction is followed by the colorimetric measurement of the rate of formation ‘of p-nitraphenot at 405 nm, which is proportional to the alkaline phosphatase activity. Absorbance data are ‘converted into 2 reportable concentration value (U/L) based on calculations for zero-order kinetics. REAGENTS The following reagents are available in the package Description ALP REAGENT When reconstituted, one liter contains: ‘p-nitrophenyl phosphate (PNPP) 16 mmol WARNING: HARMFUL IF INHALED OR INGESTED. Avoid breathing dust. Keen container closed: Use with adequate ‘enctaton ALP DILUENT ‘One liter contains: Mg* 2mmol; Zn* 1 mmol: Butter: Chelator ‘CAUTION Avoid contact with eyes, skin, fr clotting. Wash thorough" after aneting. avoid ingexton WETTING AGENT W (Prod. No. T21-1298-52) Contains: Surfactant fr clothing. Waan thorougy" ater nating. avoid ingestion Reagent Preparation The instructions for preparing the “Working Alkaline Phosphatase Reagent” vary with the specific product number of the ALP Reagents that is being used. Table 2 lists the amounts of each reagent that must be combined to prepare the “Working ALP Phosphatase Reagent." “Table 2 PREPARATION INSTRUCTIONS ” Trocaune rrosenarase] MOOT REAGENTS: ae Leena WETTING fcanron” | REAGENT mnaeNt (RCO. WO "FP ") Seg wo | amount | TORE For-reiror] Ti609 | T2187 | tome | 2OoRF frorieiaoi] Tite | T2183 | 20m | 30 roviei7an| Tiiets | T2186 | some | BOOP STORAGE AND STABILITY The ALP Reagent storage and stabil Table 3. 5 are shown in Table 3 STORAGE AND STABILITY Ses listed in Table 1 aiomae Saaury Table 1 REAGENT PACKAGING oer ‘ALP REAGENT "ALP OILUENT aman MESS, [noo vo] aucum [oaco vo, | auc youee To1-iaii-0r | Ti-te09 exiome| vero] Teo me] Torrereo [rriiesa|exzom| r-1813 fix i2smd) Bic Torte17-oW | rats [éns0me| 71-1876 [1360 mu ee ema eae ane a mites ZA Mies ine. lagnostes OWsion “Tarytows, NY 10591-5097 USA ALP (AMP)