MSACL 2016 US Abstract

Investigation of a Large Pain Profile from Fingerstick Capillary Blood

Jeffrey Enders (Presenter)
Ameritox, Ltd.

Bio: B.S./M.S. degree in chemistry from Drexel University, Undergraduate research advisor: Dr. Kevin Owens PhD in bioanalytical chemistry from Vanderbilt University under Dr. John McLean. Studied the complexation of ion mobility mass spectrometry with cell-trapping microfluidics to detect real time cellular responses.

Authorship: Jeffrey R. Enders (1), Monika Kansal (2), Yibo Guo (2), Stuart Kushon (2), Gregory L. McIntire (1)
(1) Ameritox, Ltd., 486 Gallimore Dairy Road, Greensboro, NC 27409 (2) Neoteryx, 411 Madrid Avenue, Torrance, CA 90501

Short Abstract Blood-based analysis has historically been the standard of forensic drug testing. Factors which have previously prevented blood from being the matrix of choice in pain management drug analysis, have recently been attenuated by a new sampling device which is able to collect a specific volume of blood through a fingerstick, thus negating the necessity of a phlebotomist. Commonly, fingerstick blood has been associated with various endogenous-based assays, such as glucose, hemoglobin, and genetic testing. However, in this work it will be shown that capillary blood from a fingerstick can be used to assess a patient’s exogenous compound profile.

Long Abstract

Blood-based analysis has historically been the standard of forensic drug testing. Forensic literature is replete with bloodborne metabolomics studies and statistics. Although the matrix of blood can be more difficult to prepare for typical LC-MS analyses, it is the more primary indicator of a patient’s current state when compared with the secondary excretory matrix of urine or the passive-diffusion matrix of oral fluid. Factors which have previously prevented blood from being the matrix of choice in pain management drug analysis, have recently been attenuated by a new sampling device which is able to collect a specific volume of blood through a fingerstick, thus negating the necessity of a phlebotomist. Commonly, fingerstick blood has been associated with various endogenous-based assays, such as glucose, hemoglobin, and genetic testing. However, in this work it will be shown that capillary blood from a fingerstick can be used to assess a patient’s exogenous compound profile.

Standards were prepared in donor blood using analyte Cerilliant (Round Rock, Texas) stocks. A Mitra microsampling device from Neoteryx (Torrance, CA) was used to collect exactly 10 μL of blood from either prepared standards or actual patients. On the sampling device a small absorptive bulb uses capillary action to wick the blood from the sampling surface, thus precisely collecting the target blood volume. This device was shipped (and consequently dried while on the way) to the laboratory for analysis. Sample preparation was performed by submerging the bulb in 300 μL of a methanol based internal standard. Rigorous shaking/vortexing is used to extract the compounds of interest from the bulb. 150 μL of this 300 μL extraction solvent is removed, dried down, and then reconstituted in LC starting conditions. Analysis was performed on a SCIEX 4500 triple-quad with an Agilent 1290 LC stack, using a Phenomenex (Torrence, CA) 2.6 μm phenyl-hexyl 50 x 4.6 mm column (00B-4495-E0). The fairly extensive pain panel included the following 37 compounds: Codeine, Morphine, Hydrocodone, Hydromorphone, Norhydrocodone, Oxycodone, Oxymorphone, Noroxycodone, 6-MAM, Amphetamine, Methamphetamine, Phentermine, PCP, MDA, MDEA, MDMA, Cocaine, Benzoylecgonine, Nicotine, Cotinine, 7-aminoclonazepam, Alprazolam, Clonazepam, Diazepam, Lorazepam, Nordiazepam, Oxazepam, Triazolam, Fentanyl, Norfentanyl, Tramadol, N-Desmethyl-Tramadol, O-Desmethyl-Tramadol, Buprenorphine, Norbuprenorphine, Methadone, EDDP.

Results indicated that most analytes were quantifiable down to 1 or 5 ng/mL in blood using the above detailed process and method. Matrix studies were conducted whereby, a clean standard was run to evaluate the process efficiency of the extraction. Additionally, standards were prepared at three varying hematocrit levels to assess its effect on quantitation.

The Mitra microsampling device, using only 10 μL of blood, was able to provide more than satisfactory quantitation of a large pain profile from capillary blood. Because the device is dried it minimizes the dangers inherent in handling human liquid blood and also can be shipped under ambient conditions. The ability to handle such samples in a urine drug testing facility without handling liquid blood samples affords the opportunity to provide new patient screens, illicit testing, and with further comparison testing/validation, therapeutic drug monitoring.

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