Dr. Lorna A. Nisbet Course Leader & Senior Lecturer

Transcription

1 Saliva Sampling of volunteers at London nightclubs Dr. Lorna A. Nisbet Course Leader & Senior Lecturer

2 Prevalence 2017/18 Data 19.8% of adults aged 16 to 24 had taken a drug in the last year, more than double that of the wider age group. Class A drug use increased from 6.2% in 2011/12, to 8.4% in 2017/18, Mainly driven by an increase in powder cocaine and ecstasy use. Men more likely to report (averaging double).

3 Changing Patterns

4 Oral Fluid Consists of saliva, gingival crevicularfluid, cellular debris & blood. Detection window relates to administration. Anti mortem sample used in POCT.

5 Classic Devices Until now oral fluid samples have been collected in one of two ways: 1. Raw oral fluid obtained through spitting into a collection chamber. 2. Oral fluid collected in situ onto a swab, then the swab subsequently immersed into an aqueous buffer solution then transported for laboratory analysis.

6 Oral Fluid Limitations Non-invasive sample. Longer limits of detection. (Heroin) Difficult to adulterate. Can see strong plasma: oral fluid correlation (Ethanol). Good for pharmacokinetic studies Passive contamination. Less fluid than urine. Longer sample prep time. Buffers Dry mouth

7 Issues With Oral Fluid - Buffers Stability The buffers can affect the stability of the drugs. Buffers Aqueous buffer based swab collectors were designed for now mostly redundant screening techniques such as ELISA.

8 The Device

9 Gravimetric Work 4 3,5 Saliva absorbed by swan (ml) 3 2,5 2 1,5 1 2,981 0,5 0 0,834 1,052 New device NeoSal (0.7 ml) Quantisal (1mL)

10 Extraction Efficiency

11 London Club Project-Aims 1. To match toxicological analysis of saliva samples with self-reported recreational drug use, 2. To determine the reliability of user self-report in determining the drugs that individuals are using, 3. To collect data on current patterns of drug use and awareness via self-reporting. Additionally, 4. To determine the feasibility of this new device.

12 Ethics, Health & Safety Ethical Approval - Kings College London (HR-16/ ). Ethical Approval -Anglia Ruskin's Faculty research Ethics Panel. Health & Safety Approval Anglia Ruskin Biological Safety Committee.

13 The Club Open Thursday Monday morning. Typical hours are 11pm-8am. Gay Super Club -capacity of 600. Male majority.

14 Recruitment Protocol Set up outside in the smoking area. Initially participants approached us. All recruitment done by Consultants. >5 years experience. Questionnaire. Drug use within 48 hours then sample requested. N= 31.

15 Sampling Protocol

16 Elution Protocol DOA. 2 x 2.5 ml of DCM/IPA Synthetic Cannabinoids. 2 x 2.5 ml of Hexane/EtOAc Evaporate to dryness Evaporate to dryness BSTFA PFPA:EtOAc BSTFA

17 GC-MS Instrument: Shimadzu GCMS-QP2010 Column: Zebron Capillary GC Column: ZB5 (30m x 0.32mm i.d.; film thickness 0.25µm) Carrier gas: He Injection port: 225 C Transferline: 280 C Oven temperature: 60 C Program: Initial Temperature 60 C (2 min hold) Ramp 25 C /min to 295 C (3 min hold) Total Run-time min

18 Results Cocaine Cocaine (BZE) Nicotine No Substances

19 Results Analysis Mostly male participants Sunday / Monday morning recruitment. After a bank holiday weekend Bad weather Low Participant number.

20 Limitations Fridge Storage Samples No Red Spot Mould Blood Gum No Issues

21 New Device Retained on column? Retained on swab? Sensitivity? Drugs Stability Metabolites Duration in the body

22 Future Work Match analysis with questionnaires Validate the new device. Conduct more real-life testing. Test for additional drugs. Look for adulterants.

23 Acknowledgements Duncan Carmichael Paul Dargan David Wood Andrew Scourfield Daniel Lauritzen The Volunteers

24 Questions?