:true:
The Power of Precision
false
us
ZenoTOF 7600 system
TripleTOF 6600+ system
X500R system
X500B system
View all
SelexION device
SCIEX 7500 system
SCIEX Triple Quad 6500+ system
SCIEX 5500+ system
QTRAP 6500+ system
QTRAP 4500 system
Citrine system
4500MD system
Jasper system
View all mass spectrometers
Echo MS system
Biologics Quant solution
Biotransform solution
MPX 2.0 High Throughput Multiplexing system
OptiFlow Quant solution
View all integrated solutions
BioPhase 8800 system
CESI 8000 Plus system
PA 800 Plus system
P/ACE MDQ Plus system
GenomeLab GeXP system
View all capillary electrophoresis
Advance your research with front-end instruments designed to help you realize the full power of your mass spectrometer. SCIEX has the broadest portfolio of ESI-MS front-ends that can facilitate various flow rates, sample requirements and sensitivities. No one else offers the entire range of analytical flow, microflow, nanoflow LC-MS and even ultra-low flow CESI-MS.
ExionLC 2.0 series
Micro HPLC columns
M5 MicroLC system
View All
NanoLC 400 system
Ultra-low Flow CESI-MS
View all front-end HPLC MS
Differential mobility spectrometry (DMS) and ion mobility spectrometry are analytical techniques used to separate ions based on their gas phase mobility. Multiple types of ion mobility devices exist, such as drift tubes, traveling wave, and high-field asymmetric waveform devices. Learn how you can separate yourself with Differential Ion Mobility.
The latest ion sources from SCIEX enable enhanced sensitivity and robustness with greater desolvation range across all MS platforms, from Triple Quad to QTRAP and QTOF.
Turbo V ion source
OptiFlow Turbo V ion source
OptiFlow Interface
View all ion sources
vMethod applications are pre-configured and verified LC-MS/MS methods that reduce the need for method development – significantly cutting the time, effort and money to deploy a new assay. Every vMethod provides method conditions, recommended sample prep, LC and MS conditions, and details for applicable MS/MS library databases for key applications.
AA45/20 1.0
aTRAQ
Illicit drugs
Acrylamide
Allergens
Antiobiotics/veterinary drugs
Cannabinoids
Illegal dyes
Melamine
Mycotoxins
Pesticides
Industrial chemicals (bisphenol)
Industrial chemicals (ethanolamines)
Pesticides (herbicides)
Pesticides (Polar)
Pharmaceutical and personal care products (PPCPs)
Peptide and protein bioanalysis
Routine biologics characterization
Benzodiazepines
Blood screening
Drugs of abuse
Etg and ets
Nicotine
THC-COOH
Urine screening
Explore vMethod applications
Software navigator tool
Free software trials
Software downloads
Software activation
Software support policy
Software support plans
Software feature request portal
Analyst software
Analyst TF software
SCIEX OS software
Biologics explorer software
Cliquid software
DiscoveryQuant software
Molecule profiler software
OneOmics suite
Keystone software
View all software
High resolution and QTRAP libraries can dramatically enhance the quality of your analysis, giving you much improved confidence in your data. With a comprehensive library at your fingertips, you can easily create methods and process targeted and non-targeted screening data on your complex samples, faster and easier than ever before.
All in one library
SCIEX all-in-one HR-MS/MS library with NIST 2017
Antiobiotic Llbrary
Flurochemical library
Forensic library
Mycotoxin library
Natural products
Pesticide library
Wiley Libraries
Antibiotic library
Meta library
Explore the library selector tool
Boost the performance of your mass spectrometer and improve sensitivity, productivity, and data precision. iChemistry Solutions are the world's only reagents and consumables that are custom designed with your success in mind.
RNA 9000 Purity & Integrity kit
aTRAQ kit for amino acid analysis of hydrolysates
aTRAQ kit for amino acid analysis of physiological fluids
Automated protein digestion solution
Biologics analysis kit for CE-SDS
Chemical standard kits
CYP450 protein assay - human induction kit
CZE rapid charge variant analysis kit
Fast glycan analysis and labeling for the PA 800 plus
iDQuant standards kit for pesticide analysis
Lipidyzer platform kits
roQ auEChERS extraction and dispersive kit
Ampliflex diene reagent
Ampliflex keto reagent
Cleavable ICAT reagent
iTRAQ reagent
mTRAQ reagent
View all consumables
QTOF – Quadrupole Time of Flight
QTRAP® – Triple Quad Linear Ion Trap
SWATH® – Data Independent Acquisition
SelexION® – Differential Mobility Separation
MicroLC – Microflow Chromatography
Ultra Low-Flow CESI-MS Technology
iCIEF-MS Technology
Nominal Mass LC-MS-MS
Acoustic Ejection Mass Spectrometry
View All Technology
From substantiating new discoveries through to end-to-end validated workflows for high-throughput biomarker testing in the clinic, thousands of scientists worldwide depend upon mass spec innovation from SCIEX to advance their work in the fields of clinical, research, omics and diagnostics.
Clinical Omics
Lab Developed Tests
View All Clinical
PFAS
Pesticides & herbicides
PPCP
Disinfection by-products
Soil and biota
Ethanolamine
Synthetic polymers
Exposome
Suspect screening
Nanomaterials
View All Environmental Testing
How do you protect your reputation and meet today’s global food safety standards? Whether you are a commercial lab or a food manufacturer, the quality of your food testing data is vital to your business. SCIEX solutions help you meet maximum residue limits (MRLs) with high-quality data that you can genuinely count upon. With a portfolio of applications, your lab can quickly and easily react to diverse market needs.
Pesticide Testing
Mycotoxins Testing
Antibiotics Testing
Potency Testing
Mycotoxin Testing
Terpenes Profiling
Meat Speciation Testing
Food Fraud Analysis
Food Adulterant Testing
Food Dye Testing
Food Omics
Allergen Testing
Ingredient Authenticity & Profiling Analysis
Packaging & Food Contact Substance Analysis
View All Food and Beverage Testing
How do you ensure the integrity of your results in an industry that is never constant? By accurately detecting even the smallest compound angles you can deliver evidence that stands. SCIEX forensic analysis solutions deliver fast, highly accurate data across a multitude of compounds and biomarkers, from the known to the new and novel.
Forensic Drug Testing
Designer Drug Testing
View All Forensics Testing
Discovery Proteomics
Next-Generation Proteomics
Targeted Proteomics
Discovery Lipidomics
Targeted Lipidomics
Discovery Metabolomics
Targeted Metabolomics
OneOmics
Gene Expression Analysis
DNA Sequencing
Genotyping and SNP Analysis
STR Analysis
AFLPs
View All Life Science Research
Transform the capacity and capability of your biologics pipeline with complete end-to-end solutions that make your lab more productive, and more successful. With a longstanding track record in pharma discovery, development and manufacturing, our unparalleled application knowledge with best-in-class hardware, software and support all integrate to revolutionize your lab.
Small Molecule Quant
Peptide Protein Quant
Oligonucleotide Quant
Comprehensive Metabolite Identification
Biotherapeutic Catabolism
Native Mode Analysis
High Throughput ADME
Intact Protein Analysis
Peptide Mapping
Protein Glycosylation/Microhetrogeneity
Purity/Heterogeneity Analysis
Identity/Charge Heterogeneity
Host Cell Proteins
Capsid Analysis
Nucleic Acid and Plasmid Analysis
SNP Analysis
Gene Expression Profiling
View All Pharma and Biopharma
Citrine™ Mass Spec
4500MD Mass Spec
Jasper™ HPLC System
Analyst® MD Software
Cliquid® MD Software
MultiQuant™ MD Software
ClearCore™ MD Software
Clinical Mass Spec Operators
Clinical Method Developers
Clinical Lab Managers
View All Diagnostics
The SCIEX Now Learning Hub offers the most diverse and flexible learning options available, with best-inclass content that helps you to get the most out of your instrument and take your lab to the next level. Available personalized learning paths based on the latest memory science ensure better knowledge retention, and automated onboarding and enrollment means you’ll get up and running faster.
SCIEX Now Learning Hub offers the most diverse and flexible learning options available, with best-in-class content that helps you to get the most out of your instrument and take your lab to the next level. Available personalized learning paths based on the latest memory science ensure better knowledge retention, and automated onboarding and enrollment means you’ll get up and running faster.
SCIEX Learning Manager provides you with the infrastructure to assign, monitor and report on your staff's competency through a single digital platform. Effectively manage the training process for new hires, ensure continuous staff development and access information with a single log-in to your SCIEX account.
You can browse, filter, or search our extensive list of training offerings. Choose from over 100 self-paced eLearnings or search for an instructor-led course near you. Once you select the course you want to take, you will be directed to Learning Hub for enrollment (login required).
SCIEX Now Online is the Everything, Anytime destination for all your SCIEX support needs. You can keep track of activities that matter most to you and manage your lab in the most efficient way possible. Extensive self-help resources like our deep Knowledge Base, enable you to solve many problems on your own. SCIEX Now is available 24/7 and your new SCIEX instruments are automatically added to your profile when you purchase.
Support Cases
SCIEX Now Learning Hub
Instruments
Manage My Instruments
Registered Software
Activate Software
Resource Library
My Notifications
Request Support
Course Catalog
Software Downloads
SCIEX Store
SCIEX Now New Feature Request
Software New Feature Request
Online ordering solutions
Log In
Don't have an account? Create One
Visit your SCIEX Now™ Dashboard
No one understands your needs and can support your systems better than we can. Our mission is to help you be successful, whether it's to repair your instrument, assist with your workflows, or help you maximize productivity in your lab. Whatever your challenge, global SCIEX Service and Support personnel are subject matter experts who are focused on mass spec and capillary electrophoresis, so you'll be able to achieve your scientific goals quickly and efficiently.
LC-MS Service Plans
Protect Plus Suite for Your New LC-MS
CE Service Plans
Clinical Service Plan
StatusScope Remote Monitoring
Laboratory Optimization Services
Compliance Consultancy
Qualification and Validation
On-Site Applications Support
Professional Relocation Services
View All Instrument Service & Support
Software Accelerator Program
Software Support Plans
Software Support Policy
Premium Access Content
View All Support Tools
SCIEX Now Learning Hub is much more than online courses. For the most comprehensive option, you can select Success Programs: personalized, blended online and in-person courses. If your needs are met by a visit from one of our training experts, you can choose multiple Onsite Training days to get your lab running. Visit a SCIEX training center in North America, Europe or Asia for intensive classroom and laboratory training. And your online courses are available any time, from anywhere, right here.
Login to SCIEX Now Learning Hub
Success Programs at Your Site
Online Course Catalog
Clinical Knowledge Center
Application Scientist Training at Your Site
China
Europe
German CE Courses
India
Japan
Korea
North America
UK
Visit all Training
If you have CE, LC or mass spec questions, then SCIEX has the answers. SCIEX support is the single destination for your system questions. We aim to fully assist you with virtually every application of our instruments, helping you to get the most out of your lab resources and assets.
Frequently Asked Questions
View All Request Support
As a life science researcher, you need the tools and support to help you create the scientific foundation in pursuit of expanding the knowledge-base, whether it is understanding fundamental biology, finding new biomarkers, discovering ways to improve our quality of life, or other areas of research. We are committed to the same goals and put the very same dedication into our work to help you address your most significant research challenges.
Academic Partnership Program
Academic Partners
View All Partnership Programs
Regulatory compliance is as paramount for us as it is for you. That’s why we have made it easy for you to freely reference all relevant technical and product regulatory documents. To give you confidence that, with SCIEX, you will fully comply with legislation, adhere to your laboratory protocol and meet industry standards.
Declaration of Conformance
Safety Data Sheets
Certificates of Analysis
View All Regulatory Documents
SCIEX supplies extensive documentation to help you prepare for, use, and maintain your SCIEX hardware and software products, and we update this documentation regularly. On the Customer Documents page, you can search for and download the latest documents for your product.
Customer Documents
Join the SCIEX community today to interact with your peers, share and exchange ideas, develop your knowledge, stay up-to-date with the latest products, post insights and questions, comment on others and receive support. This community is designed to help you, our customers, move science forward and get the answers you need. We’re committed to engaging with and listening to you, to create the best customer experience possible and to contribute to the success of your work.
Biopharma
Clinical
Environmental / Industrial
Food and Beverage
Forensics
Life Science Research
Pharma
Technology
Knowledge Base Articles
SCIEX Now Feature Requests
Software Feature Requests
Newsletter Archive
Featured Content
FAQs
View All Community
About SCIEX
About Danaher
Customer Profiles
Our History
Our favorite papers
Meet our executives
Career opportunities
Contact us
Press releases
In the news
Awards
You've got questions. We've got experts who can help. Contact us to find out more, talk to a specialist, explore our solutions or get expert support.
Talk to a specialist
Request more information
Request a quote
Request support
SCIEX success network
Frequently asked questions
SCIEX community
Request hosted catalog
Request punchout
Global public relations
508-782-9484
Country/Region Canada Mexico United States
Country/Region Argentina Brazil Chile Colombia Costa Rica Ecuador El Salvador Guatemala Peru Uruguay Venezuela
Country/Region Germany Albania Austria Belgium Bosnia and Herzegovina Bulgaria Croatia Cyprus Czech Republic Denmark Estonia Finland France Germany Greece Hungary Ireland Italy Latvia Lithuania Luxembourg Macedonia Montenegro Netherlands Norway Poland Portugal Romania Serbia Slovakia Slovenia Spain Sweden Switzerland United Kingdom
Country/Region Bangladesh Brunei Darussalam Cambodia 中国 Hong Kong India Indonesia 日本 한국 Singapore Sri Lanka Taiwan Thailand Viet Nam
Country/Region Bahrain Iran Iraq Israel Jordan Kuwait Lebanon Oman Pakistan Palestine Qatar Saudi Arabia Syria Turkey United Arab Emirates Yemen
Country/Region Algeria Angola Botswana Burundi Egypt Ethiopia Kenya Liberia Libya Morocco Rwanda South Africa Tunisia Uganda United Republic of Tanzania Zambia Zimbabwe
Country/Region Armenia Azerbaijan Belarus Georgia Kazakhstan Kyrgyzstan Moldova Russia Tajikistan Turkmenistan Ukraine Uzbekistan
Country/Region Australia Micronesia New Zealand
AB Sciex is doing business as SCIEX. © 2010-2018 AB Sciex. The trademarks mentioned herein are the property of the AB Sciex Pte. Ltd. or their respective owners. AB SCIEX™ is being used under license. Beckman Coulter® is being used under license. Product(s) may not be available in all countries. For information on availability, please contact your local representative. For research use only. Not for use in diagnostic procedures.
Download tech note (PDF)
Using the SCIEX Triple Quad™ 7500 LC-MS/MS System – QTRAP® Ready, powered by SCIEX OS Software
Pierre Negri1 and Ian Moore2 1SCIEX, USA; 2SCIEX, Canada
The SCIEX Triple Quad 7500 System is a robust quantitative platform with exceptional performance in speed, linear dynamic range and sensitivity for the accurate quantification of THC-COOH in oral fluid. The increased selectivity of MRM3 provided a 5 fold increase in sensitivity of detection over MRM analysis due to the enhanced selectivity of the method.
With over 192 million consumers worldwide in 2018, cannabis is the most commonly abused recreational drug.1 Detection of its use can be performed in several biological matrices such as blood, urine, hair and oral fluid. In recent years, oral fluid has gained considerable attention as a quicker and less invasive means of monitoring cannabis use. More specifically, the use of this matrix for drug testing benefits from ease of sampling, observed collection and difficulty of sample adulteration.
Δ9-tetrahydrocannabinol (THC) has previously been used as the marker of choice to monitor cannabis consumption. However, the detection and quantification of THC in drug screening can result in a high-number of false positives since this analyte can be found in subjects passively exposed to cannabis. 11-nor-9-carboxy-THC (THC-COOH) was proposed as a marker of cannabis intake since it is not detected in oral fluid collected from subjects passively exposed to cannabis.2 The low (pg/mL) concentrations of THC-COOH in oral fluid of active users together with the abundance of matrix interferences present in the stabilization buffer and oral fluid itself pose a considerable analytical challenge. In this technical note, the SCIEX 7500 System is shown to provide the required levels of sensitivity, robustness and performance for the accurate quantification of pg/mL levels of THC-COOH in oral fluid without the need for derivatization.
Figure 1. High sensitivity on the SCIEX 7500 System enabled trace level detection of THC-COOH in oral fluid. XICs showing the quantifier ion traces for the matrix blank and THC-COOH at the LLOQ for both the MRM (top) and MRM3 (bottom) workflows.
Stock solution preparation: Stock solutions of THC-COOH and THC-COOH-d3 were prepared separately in methanol at 1000 ng/mL. Blank oral fluid was spiked with the stock solution of THC-COOH to create calibrator solutions at 100000, 50000, 10000, 5000, 1000, 500, 100, 50 and 10 pg/mL. All sample concentrations are expressed as pg of THC-COOH per mL of blank oral fluid or fg of THC-COOH on column throughout this technical note, regardless of the subsequent dilutions performed.
Sample preparation: A dilute and shoot sample preparation approach was used in this experiment. In brief, 250 µL of each of the spiked oral fluid calibrator solutions was mixed with 750 µL of Quantisal stabilization buffer. The resulting solutions were then further diluted 10 times to minimize matrix effects by taking 100 µL of each solution and adding 25 µL of the 1000 ng/mL internal standard stock solution and 875 µL of a 20% methanol solution. The samples were thoroughly vortexed, centrifuged and 40 µL of the supernatant were injected for analysis. No further sample clean-up nor extraction was performed.
Liquid chromatography: HPLC separation was performed on an ExionLC™ System using a Phenomenex C18 column (50 × 2.1 mm, 2.6 µm, 00D-4475-AN) held at 40ºC equipped with a Phenomenex KrudKatcher Ultra fit (AF0-5727). Mobile phases used consisted of water, methanol and modifiers. The LC flow rate was 0.5 mL/min and the total run time was 6 min. The injection volume was 40 µL.
Mass spectrometry: A SCIEX 7500 System equipped with an OptiFlow Pro Ion Source and E Lens Technology was used. The ionization source was operated in negative electrospray ionization (ESI) mode. A looped MS experiment was performed. The first experiment consisted of five MRM transitions of 100 msec dwell time each: three for THC-COOH and two for THC-COOH-d3. The second experiment was an MRM3 scan using a scan speed of 10000 Da/s with 200 msec fill time, 25 msec excitation time and an 8V Q3 entry barrier. MRM3 data was collected as full scan data from 180 to 250 Da to simultaneously monitor the 343.1 -> 299.2-> 244.9 and the 343.1 -> 299.2 -> 191.1 MRM3 transitions. The Guided MRM3 automated compound optimization feature in SCIEX OS Software was used for optimizing the parameters for the MRM3 experiment. Six replicates of each sample were injected to build a data analysis processing method.
Data analysis: Data processing was performed using SCIEX OS Software. Quantitative analysis was performed using AutoPeak Algorithm in the Analytics module of the software where calibration curves, concentration calculations, assay precision and accuracy statistics were generated.
Accurate quantification of meaningful (pg/mL) THC-COOH levels in human oral fluid is paramount to confirm THC consumption. However, the low concentration of THC-COOH found in oral fluid of active THC users and the complexity of this matrix make detection of this metabolite challenging. As a result, developing a sensitive workflow that can accurately quantify trace levels of THC-COOH in oral fluid is critical to confirm active cannabis consumption.
The ability to accurately detect low concentration levels of THC-COOH extracted from human oral fluid was assessed using the SCIEX 7500 System. Six replicates of each of the processed calibrator samples were injected to determine the lower limit of quantification (LLOQ) value for THC-COOH and to assess the overall robustness and reproducibility of the instrument. The LLOQ value for THC-COOH was determined to be the lowest concentration calibration level meeting the following analytical performance requirements: signal-to-noise ratio (S/N) ≥10, calculated concentration accuracy within 15% of 100%, precision (%bias) below 10%, and calibrators falling on a linear calibration curve with an R2 value of at least 0.99. Detection and integration of the peaks was automatically performed using the AutoPeak Algorithm in the Analytics module of SCIEX OS Software. Analyte concentration and ion ratio were calculated automatically in the software.
Figure 2 shows the extracted ion chromatogram (XIC) traces and resulting calibration curves for two of the MRM transitions monitored for THC-COOH. The two series of XIC traces for both the quantifier and qualifier ions of THC-COOH showed a high level of sensitivity and precision across concentrations ranging from 50 to 100000 pg/mL. Eight levels of calibrators were used to determine the ion ratio criteria for the quantifier and qualifier ions of THC-COOH. The results demonstrated excellent correlation of the generated regression curves covering concentration ranges meeting the requirements of pg of THC-COOH per mL of blank oral fluid: lower limits of quantification (LLOQ) was determined to be 50 pg/mL (50 fg on column) for the quantifier ion. In addition, excellent linearity was observed across the concentration range from 50 to 100000 pg/mL for the quantifier ion and from 100 to 100000 pg/mL for the qualifier ion, with R2 values of 0.99870 and 0.99868, respectively.
Figure 2. High sensitivity and linearity using MRM for the detection of THC-COOH in oral fluid. (A) Calibration curves resulting from the calibration series for THC-COOH from 50 to 100000 pg/mL and 100 to 100000 pg/mL for the quantifier and qualifier ions, respectively. (B) Selected XIC traces of the quantifier and qualifier ion for the matrix blank and THC-COOH at 100, 1000, 10000 and 100000 pg of THC-COOH per mL of blank oral fluid. A total of six injections were performed for each calibrator sample.
A series of six replicate injections were performed to evaluate the robustness and overall performance of the SCIEX 7500 System. Table 1A summarizes the MRM results from the average (n=6) injections of each of the calibrator solutions spiked with THC-COOH and extracted from human oral fluid. The assay showed excellent precision and accuracy for concentrations ranging from 50 to 100000 pg/mL for the two THC-COOH MRM transitions monitored. All quantified samples had a %CV value below 10% and accuracy values ranging between 91.38 and 111.38%. These results demonstrate that the SCIEX 7500 System can provide robust and reproducible results even for challenging forensic workflows requiring detection of extremely low concentration levels of analytes. The performance showcased by the SCIEX 7500 System prove to be adequate to achieve the required levels of sensitivity, robustness and reproducibility required by the current workflow to accurately quantify THC-COOH down to pg/mL levels.
Table 1. Statistical results table generated in SCIEX OS Software showing accuracy and precision of THCOOH for the series of calibrator samples using (A) MRM and (B) MRM3. Average (n=6) results generated in Analytics show high levels of precision (<10%) and accuracy (within 15% of 100%) for concentrations across the respective calibration curves for the quantifier and qualifier ions for both experiments. Reported concentrations are pg of THC-COOH per mL of blank oral fluid without accounting for the subsequent dilutions performed during the sample preparation procedures.
Detecting low amounts of a metabolite present in complex biological specimen without extensive or time-consuming sample cleanup is often prone to high background and matrix interferences. In such cases, accurate quantification of specific analytes using MRM can be impaired due to the high matrix background. What typically ensues is a compromised LLOQ as the detection of these analytes is limited by signal-to-noise rather than raw instrument response. The addition of a third MS stage combining normal LIT functions with normal quadrupole functions using a novel QQQ/LIT workflow coined MRM3 can significantly reduce these background interferences while providing increased selectivity and sensitivity at the low end of the calibration curve.3 The principle of MRM3 analysis as performed on the SCIEX Triple Quad 7500 System – QTRAP Ready is shown in Figure 3.
The MRM3 quantification assay was performed on the same set of calibrator samples. Two secondary product ions of the 343.1à299.2 transition were isolated, fragmented in the linear ion trap, extracted using a width of 1 Da and summed together to produce the MRM3 signal. Figure 4 shows detection of THC-COOH at 50 pg/mL using MRM (top left) and MRM3 (top right). However, due to higher background when running the MRM assay, no signal was observable at 10 pg/mL THC-COOH (bottom left) in either of the two MRM transitions monitored. MRM3 showed much better specificity and sensitivity than the MRM data at this concentration (bottom right). As seen in Figure 4, the use of the MRM3 workflow provided much higher sensitivity compared to MRM alone due to the additional level of selectivity obtained by monitoring secondary product ions for THC-COOH in oral fluid.
Figure 3. MRM3 scan mode for quantitative analysis by LC-MS. Analyte’s precursor ions are selected in the Q1 quadrupole, fragmented in the Q2 collision cell before being collected in the linear ion trap (LIT) in Q3 where a suitable fragment ion is successively being isolated then fragmented using resonance excitation. These second generation product ions are then collected out of the LIT and scanned by the detector, enabling higher specificity analyte detection with better sensitivity from highly complex biological samples.4
Figure 4. Comparison of MRM vs. MRM3 for the detection of THC-COOH in oral fluid. (Top) Both MRM (left) and MRM3 (right) workflows enabled detection and quantification of THC-COOH at 50 pg/mL in oral fluid. However, the MRM3 workflow enabled significant reduction of background caused by matrix interference, significantly improving selectivity and resulting in a better LLOQ. (Bottom) Using MRM, THC-COOH was not detectable at 10 pg/mL due to higher background (left) however the MRM3 approach provided good detection of THC-COOH at this level (right).
The LLOQ for the MRM3 was also determined by processing the MRM3 data in SCIEX OS Software. Figure 5 shows the extracted ion chromatogram (XIC) traces and resulting calibration curves for the two secondary product ions monitored for THC-COOH. The LLOQ was determined to be 10 pg/mL (10 fg on column) for the quantifier ion. Calibration curves were also generated for the MRM3 workflow and showed excellent linearity across the concentration range from 10 to 10000 pg/mL for the quantifier ion and from 50 to 10000 pg/mL for the qualifier ion, with R2 values of 0.99782 and 0.99648, respectively. Overall, a 5-fold improvement in LLOQ was observed for the MRM3 experiment (10 pg/mL) compared to the MRM experiment (50 pg/mL). While MRM detection suffered from high background caused by matrix interference at low concentrations, MRM3 detection resulted in more selectivity and improved the detection sensitivity. Table 1B summarizes the MRM3 results from the average (n=6) injections of each of the calibrator solutions. The precision values (%CV) were <10% with accuracies between 87.43 and 109.49 for concentrations ranging from 10 to 10000 pg/mL for the quantifier ion and from 50 to 10000 pg/mL for the qualifier ion, respectively.
Figure 5. Significant improvement in LLOQ using MRM3 for the detection of THC-COOH in oral fluid. A) Calibration curves resulting from the calibration series for THC-COOH from 10 to 10000 pg/mL and 50 to 10000 pg/mL for the quantifier and qualifier ions, respectively. (B) Selected XIC traces of the quantifier and qualifier ion for the matrix blank and THC-COOH at concentrations at the low end of the curve (10, 50, 100 and 500 pg of THC-COOH per mL of blank oral fluid). A total of six injections were performed for each calibrator sample.
A highly sensitive workflow for the detection of THC-COOH in oral fluid is described using the SCIEX Triple Quad 7500 System - QTRAP Ready.4 The use of a rapid and simple dilute and shoot approach without the need for sample pre-treatment or derivatization provided a comprehensive extraction method that is suitable for high-throughput oral fluid drug testing in the forensic toxicology laboratory. The high sensitivity of the SCIEX 7500 System enabled accurate and sensitive quantification of THC-COOH in oral fluid with an LLOQ of 50 pg/mL (50 pg on column). This unparalleled sensitivity was achieved with excellent linearity and without any sacrifice or compromise in data quality, as demonstrated by the excellent precision (<10%) and accuracy (between 91.38 and 111.38%) across the calibration range (50 to 10000 pg/mL). The MRM3 workflow was shown to increase selectivity by significantly reducing background and matrix interference, resulting in a 5-fold improvement in LLOQ compared to the MRM approach. The results of the MRM3 experiment also demonstrated excellent linearity, precision and accuracy across the calibration range.
Overall, these results demonstrate that the levels of sensitivity and robustness achieved by the SCIEX 7500 System enable accurately quantification of low levels of THC-COOH in oral fluid without the need for time-consuming sample preparation or derivatization. Therefore, the presented workflow is readily adaptable for high-throughput toxicology testing and provides a reliable and sensitive means of differentiating passive environmental exposure from active cannabis consumption.
We use cookies to optimize site functionality and give you the best possible experience.