Webinar

Please mute your microphone

Routine Analysis of Narcotics Drugs on Thermo Scientific GC-MS

Hendy Dwi Warmiko

Sales Support Spv
Tuesday, Dec 15, 2020
PT GeneCraft Labs
1 The world leader in serving science
2
Drug of Abuse Testing?

GC-MS
LC-MS/MS LC-HRMS

Rapid Test

3
Sample Preparation

4
Objectives of Sample Preparation

5
Comparison of LLE with HyperSep Verify CX SPE

6
HyperSep Silica Based SPE

7
ISQ-7000 GC-MS

Unstoppable robustness and productivity

offers innovative design capabilities for increased
productivity and robustness, with enhanced easier
customer experience and unmatched sensitivity for the
most challenging applications

Instant access for ease of use

TRACE 1310 GC offers local control fatures a complete
icon-driven touch-screen user interface ideal for direct
instrument control in larger routine and method
development laboratories.

Surpass competition in the GCMS market with leading performances and scalability

8 Proprietary & Confidential

 GC Equipment Design since 1955…
Pneumatic control
• Diverse inlet and detector types are
Inlet/
required to run different GC applications detector
bodies Specific
• Inlet and detector bodies are installed on electronic
control
oven top deck and require pneumatic and boards
specific electronics controller
Inlet/
• The large number of options can sum up detector
bodies
to thousands of possible combinations
and final system configurations
• Typically systems are factory-configured
based on orders
• Upgrades or changes in configuration at
site is an expensive, difficult and time-
consuming operation GC oven

9 Proprietary & Confidential

 A new Modular Approach to GC Instrumentation design
Pneumatic control Electronic boards Proportional solenoid valves
for gas control
Inlet/
detector
body Specific
electronic
control
boards Pneumatic manifold

Inlet body heater and Insulation

Column connection
(inside the oven)

GC oven

• Instant connect modules incorporate:

• Injector (or detector) body and heater
• Gas control Valves
• Electronics for temperature and gas control, for signal amplifier and A/D conversion

10 Proprietary & Confidential

ISQ 7000 GCMS – Designed with Intention

Advanced Electron Ion (AEI) source

(available in dedicated configurations)

Quadrupole Mass Analyzer

S- shaped ion guide

Dual Filament
assembly

Triple off-axis DynaMax XR

detection system

ExtractaBrite ion source

Vacuum Probe
Interlock (VPI)
(Optional)

11 Proprietary & Confidential

NeverVent technology for no vent source exchange

Extends the capability of the vacuum probe interlock (VPI)

design with the newly introduced source plug, V-Lock

Through the VPI, no need to vent mass spec system for

extracting the wireless ExtractaBrite ion source

Step 1. Insert removal tool Step 2. Remove source Step 3. Hot source is held in tool Step 4. Push source out of tool

12 Proprietary & Confidential

NeverVent technology for column exchange
What is NeverVent technology?

Extends the capability of the Vacuum Probe Interlock (VPI) design with
the newly introduced source plug, V-Lock

Through the VPI and the V-Lock source plug, no need to

vent mass spec system to change the analytical column
V-Lock
Isolate the MS under
vacuum from the GC
Through the VPI and the V-Lock source plug, no need to
No complicated fluidics
or extra connections vent mass spec system to make injector maintenance

13 Proprietary & Confidential

Unstoppable Uptime

Time saving by using the NeverVent technology

Exchange ion source Column Replacement

(including conditioning)

4 hours
98% 4.5 hours
87%
Time Time
98% Saving 87%
Saving
Time Time
Saving Saving

14 Proprietary & Confidential

Technical Note 10592

15
Technical Note 10592

The urine samples were subjected to a solid phase

extraction procedure:

• 30 μL -glucuronidase (Merck 5000 I.U.) were added
to 3 mL of urine and incubated for 60 min at 56 °C.
• The Thermo Scientific™ HyperSep™ Verify CX cartridge,
6 mL/200 mg, was conditioned with 3 mL MeOH
followed by 3 mL 0.1% formic acid.
• Urine was mixed with 3 mL of 2M acetate buffer, pH
4.8. Urine was checked and adjusted, where
necessary, for accurate pH.
• The sample was added to the HyperSep Verify CX
cartridge and a slight vacuum was applied to achieve,
for example, approximately one drop per second
elution rate.
• Interference elution was done with a mixture of 1 mL
water + 0.1% formic acid, total volume 3 mL, followed
by a mixture of 1 mL MeOH/water 50:50 + 0.1% formic
• acid, total volume 3 mL.
• The cartridge was dried after interference elution with
strong vacuum.
• Elution was performed with a mixture of methanol and
5% ammonia solution (95:5), pH 9, two times 0.5 mL.
• The sample was evaporated under nitrogen until
dryness at 65 °C.
• The sample was reconstituted with 50 μL MeOH and
placed in 50 μL MS certified vials.
The vials were subsequently centrifuged for precipitating
the particles before putting them in the autosampler.

16
Technical Note 10592

• Thermo Scientific™ HyperSep™ Verify CX Cartridges, 200 mg/3 mL; 50 pack (P/N 60108-777)
• 24-port Vacuum Manifold (P/N 60104-233)
• Vacuum Pump (P/N 60104-233)
• Analyte Elution 4 mL Vial
• Thermo Scientific™ Reacti-Vap™ Evaporator (P/N TS-18826)
• Thermo Scientific™ SureStop™ MS Certified Vials, 100 μL reservoir (P/N MSCERT5000-36LVW)
• Thermo Scientific™ Marathon™ Injection Port Septa (P/N 313P3240)
• Thermo Scientific™ LinerGOLD™ GC Liners (P/N 453A-1255-UI)
• Thermo Scientific™ TRACE™ TR-DoA 35MS column, 15 m, 0.25 mm ID, 0.25 μm (P/N 26AF130P)

17
Technical Note 10592
Compound separation and detection was achieved
using a Thermo Scientific™ TRACE™ 1310 GC coupled
with a Thermo Scientific™ ISQ™ 7000 mass spectrometer
Sample introduction was performed using a Thermo
Scientific™ TriPlus™ 100 LS autosampler, injecting 1 μL into
the Thermo Scientific™ Instant Connect Split/Splitless
(SSL) injector module.
Tables 1 and 2 list the method parameters.

18
19
20
ISQ 7000 single quadrupole GC-MS system

Ultimate sensitivity with Electron Ionization source

Incredible uptime with inherent robustness and NeverVent™ technology

Routine ease of use from method development to daily operation

True scalability for growing laboratory requirements

21
Thermo Scientific GC Sampling Systems
Thermo Scientific™
Thermo Scientific™ Thermo
Scientific™ TriPlus™ RSH™
TriPlus™ 100 LS
TriPlus™ 500 Autosampler
Headspace
Autosampler

Sample Throughput
Automation

120/240 vial
configuration

Thermo Scientific™
AI/AS 1310 Series
Autosampler 12 vial TriPlus 500 HS
configuration + AI/AS 1310

Liquid Injection Headspace Injection All in One

22
Thermo Scientific Chromeleon CDS software

Chromeleon CDS software

• Control your entire chromatography lab. It is fully scalable from a single
workstation to an enterprise-wide installation

• Control more than 350 modules from Thermo Fisher Scientific™ and many

other vendors

• Quantitative mass spectrometry workflows for all separation

techniques and MS variants, all using the same intuitive user interface

• Boost laboratory efficiency with operational simplicity and intelligent

functionality

23
Expanded Sympathomimetic Amines by GC-MS

HyperSep™ Verify-CX solid phase extraction

columns (200 mg, 10 mL, P/N: 60108-742) were
used for sample extraction, and extracts were
derivatized with trifluoroacetic acid anhydride
(TFAA). Ethyl acetate was used to bring the final
extract volume to 100 μL.

24
THC Metabolite in Urine by GC/MS

All validation studies were prepared in urine using a 3 mL

sample size and derivatized with BSTFA with 1% TMCS
(Thermo Scientific, P/N: TS-38834). HyperSep™ Verify- TraceGOLD TG-DoA35MS column
CX solid phase extraction columns (200 mg, 10 mL, P/N:
60108-742) were used for sample extraction. THCA-D9
was used as the internal standard.

25
Codeine and Morphine

26
Codeine and Morphine by GC-MS

27
Ephedrine and Pseudoephedrine by GC-MS

28
Benzoylecgonine in Urine by GC-MS

Thermo Scientific HyperSep Verify-CX solid phase

extraction columns (200 mg, 10 mL, P/N: 60108-742)
were used for sample extraction. Samples were
derivatized with hexafluoroisopropanol (HFIP) and
pentafluoropropionic acid (PFPA or PFAA).

29
Testing Drugs in Hair

The hair sample must be collected a minimum of 4-6

weeks after the alleged incident. This period is adequate
to ensure that the hair shaft incorporating the drug
emerges from the bulb area in the follicle to a height
above the skin surface sufficient for collection.

The victim must avoid cosmetic treatments or haircuts

until the investigation has been concluded.

30
Hair Sample Preparation by GC-MS (UNODC General Method)

Decontamination Clean-up
Procedure with organic solvent Treat 30-50 mg of washed hair by extraction or digestion/extraction as
1. Take a hair strand (~ 100 mg). described above. Extensively centrifuge the extracts
2. Wash with 5 ml dichloromethane for 2 min.
3. Dry with adsorbent paper. SPE clean-up
4. Wash again in 5 ml dichloromethane for 2 min. 1. Condition an SPE column with methanol (2 ml) and phosphate
5. Dry again. buffer 0.1 M, pH 6 (2 ml).
2. Transfer the supernatant hair extract to the column, taking care to
Extraction avoid transferring the precipitate.
With methanol or aqueous acids or buffered solutions. Incubation in 3. Wash with 2 ml of water.
aqueous 0.01-0.50 M HCl or in 1 M phosphate buffer at pH 6.4-7.6 4. Wash with 3 ml of 0.1 M HCl.
is usually performed at 56°C or 60°C overnight. 5. Dry the column at maximum vacuum for 10 minutes.
6. Wash with 5 ml of methanol.
Digestion 7. Elute with 2 ml CH2Cl2/Isopropanol (80:20) at 2 % of NH4OH.
With diluted NaOH, 1 M NaOH is added to hair and incubation is typically 8. Dry the organic eluate under nitrogen at 40°C; if gas
performed for one hour at 80°C, or overnight at 60°C. It is suitable only for chromatographymass spectrometry (GC-MS) is planned (see
drugs that are stable under alkaline conditions. below), derivatize by adding N-Methyl-N-
(trimethylsilyl)trifluoroacetamide (MSTFA) + 1 % of
trimethylchlorosilane (TMCS) at 75°C for 20 minutes).

31
Application Note 10493

32
Application Note 10493

33
Application Note 10493

34
Technical Note 10592

Over the course of several weeks, over 700 urine samples were analyzed

35
Technical Note 10592

36
Technical Note 10592

37
Technical Note 10592

38
Thermo Scientific HyperSep SPE Applications Example

39
GC-MS for Forensic Analysis

• Used extensively for decades in analytical forensic toxicology laboratories, “gold standard”
• All analytes eluting from column are hit with a beam of electrons at 70 eV so fragmentation pattern is
identical on all instruments
• Published in a searchable full scan GCMS libraries (NIST, SWGDRUS, AAFS, Maurer Wiley)
• Analysis performed in a gas phase
• To maximize sensitivity, targeted analysis can be conducting using SIM (selected ion monitoring)
• Straight forward, simple operation
• Sensitivity ug/mL to ng/mL
• Cheaper than LC-MS/MS or LC-HRMS

40
Comparison of GC-MS, LC-MS/MS, and LC-HRMS

GC-MS LC-MS/MS LC-HRMS

Screening Full Scan, Targeted Targeted Full Scan, Targeted
Analytes Small, volatile, Small or large, non- Small or large, non-
thermally stable, volatile, thermally volatile, thermally
non-polar unstable and polar unstable and polar
Resolution 1 amu 1 amu 0.0001 amu
Libraries Large, transferable Smaller, non- Smaller, non-
transferable transferable
Analyte EI full scan spectra MS/MS Exact mass, isotope
identification and retention time fragmentation and pattern,
retention time fragmentation and
retention time
Cost Rp RpRp RpRpRp

41
Library (GC-MS and LC-HRMS)

LC-HRMS (Orbitrap)

GC-MS

42