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Eas 2011 Fei Invited Sers Talk Drugs In Saliva
 

Eas 2011 Fei Invited Sers Talk Drugs In Saliva

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    Eas 2011 Fei Invited Sers Talk Drugs In Saliva Eas 2011 Fei Invited Sers Talk Drugs In Saliva Presentation Transcript

    • Rapid Detection and Identification of Drugs in Saliva by Surface-Enhanced Raman Spectroscopy Frank E. Inscore, Chetan Shende, Atanu Sengupta, Hermes Huang and Stuart Farquharson Focus: SERS Applications in Biofluids Screening Abused Drugs in Saliva at Road-Side & Emergency-Room for Driver Impairment & Overdose Relevant R&D Funding to RTA: NIH CN: 1R43CA94457-01 NSF CN: DMI-0215819 NASA CN: NNC05CA09C Jet Propulsion Laboratories (Dr. Eric Wong) UK Road Policing Technologies Home Office Scientific Development Branch (Dr. Helen Turner, Dr. Audrey Carmichael)www.rta.biz860-635-9800 EAS 2011inscore@rta.biz Providing Chemical Information When & Where You Need It RTA Booth #106
    • The Need & Challenge: Drugs in salivaAnalysis of chemo-drugs and metabolites for dosage control• Dosage critical (to little noneffective, to much kills patient)• Current analysis requires large sample volume (10-20 ml blood and/or urine)• Traditional lab methods are labor intensive and time consuming• Viable alternative is saliva, parent drugs/metabolites adequately represented (but at lower levels)• Advantages of saliva, non-invasive (no needles) & 100X less potential interferents (99.5% water)Road-side screening for determining drug induced driver impairment National Highway Traffic Safety Administration Stats:• 2007: 11% of drivers stopped tested positive for drugs• 2009: 18% of driver fatalities tested positive for drugsEmergency room assessment of overdose and drug abuse U.S. Drug-Related Emergency Room (ER) Stats:• 2004: 2.4 million visits• 2009: 4.6 million visits 2.1 million attributed to illicit drugs Cocaine, Heroin, Methamphetamine, PCP, MDMA, LSD, Methadone 2.3 million attributed to prescription & some to over-the-counter (OTC) drugs Oxycodone, Diazepam & Acetaminophen Providing Chemical Information When & Where You Need It
    • Challenge: Drugs in salivaCritical need to rapidly identify offending drug(s), so that impaired drivers can bearrested or appropriate medical care can be administered.The analyzer must typically provide the following criteria: • Specificity – Identify and Discriminate Drugs (with No False Positives!) • Sensitivity – Detect ~10-8 M or less (e.g. 30 ppb cocaine is threshold) • Reproducibility – Accurate and Repeatable (with No False Negatives!) • Speed – Rapid Response and Analysis within 8-10 minutes • Field Usable – Battery Operated and Rugged In an effort to meet this need we have been investigating the ability of surface- enhanced Raman spectroscopy (SERS) to detect and identify numerous drugs of abuse in saliva at ng/mL concentrations within 10 minutes. We present successful measurement of representative illicit, prescribed, and over-the-counter drugs in saliva by SERS, with a focus on cocaine. Providing Chemical Information When & Where You Need It
    • The Solution: SERSSpecificity: all chemicals (drugs/metabolites) produce a unique Raman spectrumallowing unequivocal identification (no false-positives).Sensitivity: Ag and Au nanoparticle substrates used to generate SERS amplifyRaman signals (increase scattering efficiency) by 1 million times or moreallowing required detection of 10-8 M (ppb) (no false-negatives). CH3 + H Cl - N CO2 CH3 Raman: O2 C Pure Cocaine Cocaine Hydrochloride Au Gold SERS: Ag 1 ppm Cocaine Providing Chemical Information When & Where You Need It
    • How it works: RamanLight Chemical virt H H Transmittedhνo H H hνvib Absorbed (IR) hνo hνscat H H Raman Raman Scattered hνscat vib1 Rayleigh hνvib vib0 Laser light directed at a chemical generates Raman light. Providing Chemical Information When & Where You Need It
    • Surface-enhanced Raman SpectroscopyRaman, although weak effect, provides molecular specificity hν Ag BUT, when a molecule is within a laser induced plasmon field, H H N N H N N H SERS N Hthe efficiency of Raman scattering can Plasmon Field provides increase by 106 i.e. 1 million times! Surface-Enhanced Single Molecule Detection Raman PhotonSub part-per million detection possible some argue this requires enhancement factor (EF) ALSO, chemical contribution ofcan provide additional 103 enhancement 1012 -1014Sub part-per-billion detection becomes possible with SERS Providing Chemical Information When & Where You Need It
    • SERS-Active Media Traditional: • Electrochemically Roughened Electrodes • Metal Colloidal Hydrosols • Metal Islands or Nanoparticles on Solid Supports • Metal Coated Surface Structures • Self-Assembled Monolayers (SAMs) Recent: • Metal-Doped Porous Media • Periodic Apertures in Metals • Metal Shells • Fiber Optic Tips Providing Chemical Information When & Where You Need It
    • Commercial SERS Substrates: Benzenethiol benzenethiol Conc. EF (A) LOW SENSITIVITY SLOW RESPONSE 10-3M 102 BUT REPRODUCIBLE (B) LESS REPRODUCIBLE 10-5M 10 4 SLOW RESPONSE BUT MODERATE SENSITIVITY (C) HIGH SENSITIVITY AND FAST RESPONSE 10-8M 107 (~10ppb) AND OK REPRODUCIBLE RTA: LMC ~10-11M (0.01 ng/mL or 10 ppt) Providing Chemical Information When & Where You Need It
    • Approach: RTA SERS Patented Sampling Systems provide instant response in seconds as opposed to 30-min or more! 2001: Simple SERS Sample Vials Molecules Sol-Gel Matrix Raman in Solution Scattering Laser Adsorbed Molecules Metal Particle silver gold 2003: SERS Microplate 2004: SERS-Active Capillary 1 10High Throughput Screening Extraction and Pre-Concentration U.S. Patents for RTA6,623,977; 6,943,031; 6,943,032; 7,312,088; 7,393,691; 7,393,692; 7,462,492; and 7,462,493 Providing Chemical Information When & Where You Need It
    • 2007: Functionalized Sol-Gel SERS Capillary (affords greater selectivity and sensitivity) PC Std chromatographic media OTC Providing Chemical Information When & Where You Need It
    • R&D: SERS Lab-On-ChipDifferent wafer, glass and plastic Lab-on-Chip designs used Providing Chemical Information When & Where You Need It
    • RTA’s SERSID - Trace Chemical Analyzer’s for Field and Lab Use 2010 2011 Patents: 6623977, 6943031, 6943032, 7312088, 7393691, 7393692, 7462492, 7462493, 7713914 Providing Chemical Information When & Where You Need It
    • The Proposal: The Device The proposed SERS-RSSD-DD (Road Side Screening Device for Drug Detection) and SERS-ERSD-DD(Emergency Room Screening Device for Drug Detection) will extract, identify, and quantify the presence of drugs (and metabolites) in driver or patient saliva at ~10-8M within 8-10 minutes. 6 Providing Chemical Information When & Where You Need It
    • UK Road-Side Screening: Required Drugs 35 drugs & metabolites were measured31 active on gold, 4 (barbiturates) active on silver Spectra search worked for all drugs Providing Chemical Information When & Where You Need It
    • 150 Drugs in Expanded Gold SERS Library: Applicable to the ER See recent RTA paperPharmaceutics 2011, 3, 425-439doi:10.3390/pharmaceutics3030425Providing Chemical Information When & Where You Need It
    • SERS of Representative Illicit, Prescription & OTC Drugs in Expanded Gold Spectral Library (150) PCP Diazepam Methamphetamine Ritalin MDMA Demerol LSD Hydrocodone Heroin Oxycodone Acetaminophen Acetylsalicylic acid Ibuprofen Providing Chemical Information When & Where You Need It
    • Library Search: Measure Priority Drugs Spectra search and identification worked for all drugs Hit Quality Name 1 0.001 Nordiazepam 2 0.010 Methadone 3 0.010 Oxazepam 4 0.010 Temazepam 5 0.012 Norcodeine Hit Quality Name 1 0.036 Nordiazepam 2 0.357 Temazepam 3 0.363 Diazepam 4 0.373 Methadone 5 0.392 Oxazepam Providing Chemical Information When & Where You Need It
    • Spectral Match Results for Mixtures500ppmnordiazepam /diazepamDiazepam RefNordiazepam Ref 500ppm cocaine/diazepam Diazepam Ref Cocaine Ref
    • Cocaine: Static Concentration Data 50 ppb Cocaine Background = Luminescence25 ppb = 7x10-8M from glass capillary It can be subtracted. Providing Chemical Information When & Where You Need It
    • Sensitivity: Static ROC Curves of Cocaine 50 ppb 100 ppb 25 ppb 75 ppb 50 ppb 25 ppb 0 ppb Conc. # of Mean Std Mean Std Log C at LMC at substrates peak hgt Deviation Deviation (α) Conc. C K value K=3.29 95 % Blank 10 0.002 0.0014 0.004 25 ppb 2.263 confidence 25 ppb 9 0.0113 0.006 50 ppb 3.343 50 ppb 10 0.0157 0.003 -7.31583 48 ppb New Modified Gold Sol-Gel ROC Curves 95% Confidence = 48 ppb (10 capillaries measured at 9 spots per concentration) Providing Chemical Information When & Where You Need It
    • Improve sub-ppb Detection via Pre-concentrationflowed 5 mL of Sensitivity and Reproducibility25 ppb cocaine in water achieved with flowing less than 5 mL of 25 ppb (10-8M) cocaineHCl in HPLC water on new gold SG4(3H); A detected at 95% confidence (K > 3.29) in under 5-min! B Also static 25 ppb detected 25 ppb cocaineHCl on gold SG4 after extracted cocaine in water from 1 mL HPLC water with SPE capillary column 8-min; intensity 40x the static signal and is equivalent to a 1ppm signal as observed on the static concentration curve. extracted 5 mL of 25 ppb (7.3x10-8M) cocaineHCl 25 ppb cocaine in water 12.5 ppb (9.2x10-8M) amphetamine static 25 ppb 25 ppb (8.7x10-8M) diazepam cocaine in water Providing Chemical Information When & Where You Need It
    • Develop Saliva Extraction Method 1. Collect 0.5 ml in 1 min 2. Expel sample into buffer 3. Filter sample4. Draw sample into microSPE 5. Elute cocaine from SPE 6. Inject sample in SERS Capillary 7. Measure spectrum Providing Chemical Information When & Where You Need It
    • Cocaine Extracted from Saliva25ppb cocaine in 0.5mL saliva extracted & detected in 8-min (from start to finish) 25 ppb Cocaine Amphetamine in 0.5 ml Saliva Diazepam Methadone PCP 25 ppb Cocaine in 0.5 ml Water500 750 1000 1250 1500 1750 Wavenumbers (cm-1) Above drugs at 1ppm extracted from 0.5 mL saliva in 8-min Providing Chemical Information When & Where You Need It
    • Spectral Search and Match Results for Drugs in Saliva 50 ppb cocaine 100ppm Oxycodone 1ppm PCP Lib. Oxycodone 1ppm diazepam 10ppm acetaminophen Lib. Hydrocodone Unknown Oxycodone Cocaine PCP Diazepam Acetaminophen (0.1 mg/mL water) (50 ng/mL) (1 mcg/mL) (1 mcg/mL) (10 mcg/mL) Rank HQI Chemical HQI Chemical HQI Chemical HQI Chemical HQI Chemical 1 0.073 Oxycodone 0.287 Cocaine 0.019 PCP 0.022 Diazepam 0.276 Acetaminophen 2 0.734 Hydrocodone 0.348 Ethyl- benzoyl-ecgonine 0.315 Fentanyl 0.317 Temazepam 0.639 Sulfadoxine 3 0.800 Trazadone 0.349 Benzoyl-ecgonine 0.325 EMDP 0.328 Nor-diazepam 0.704 Serotonin Providing Chemical Information When & Where You Need It
    • Further Test: 50ppb Drug in Saliva Samples Cocaine 50ppb Hit Quality Name 1 0.236 Cocaine Diazepam 50ppb 1 0.171 Diazepam PCP 50ppb 1 0.171 PCP Methadone 50ppb 1 0.066 Methadone Providing Chemical Information When & Where You Need It
    • Drug in Saliva Extraction: Potential Interferents 50ppb in saliva via SPE method Caffeine Quinine vitC Mannitol50ppb equal mixturecocaine and caffeineextracted from 0.5mLsaliva in 8-min 30ppb cocaine and 50ppb mannitol mixture extracted from 0.5mL saliva in 8-minblank saliva extracted as control Correctly Identified Cocaine by Spectral Match! Providing Chemical Information When & Where You Need It
    • Preliminary LOC Designs Plastic Cover SPE PMMA with Ag/Au Channels Glass PlateProviding Chemical Information When & Where You Need It
    • Preliminary LOC Results: in Saliva at 50 ppb amphetamine 50ppb diazepam 50ppb cocaine 50ppb PCP 50ppb methadone 50ppb All Drugs Correctly Identified by Spectral Match! Providing Chemical Information When & Where You Need It
    • Next Generation LOC Design Buffered 3mL syringe saliva sample Lab-on-chip SPE capillary SERS capillary 0.2µm filter Elution solvent Valve reservoir Valve Valve Solvent waste reservoir Vacuum Sample waste reservoir Providing Chemical Information When & Where You Need It
    • SummaryCritical need to rapidly identify offending drug(s) in impaired drivers and overdose in ER.Demonstrated ability of surface-enhanced Raman spectroscopy (SERS) to detect and identifynumerous drugs in saliva at ng/mL concentrations within 8 minutes. Identification is provided by matching measured spectra to a SERS library comprised of over 150 different drugs, each of which possess a unique spectrum. Trace detection is provided by gold nanoparticles trapped within a porous glass matrix that generate SERS. Speed is provided by a syringe driven sample system that extracts the drugs from saliva AND provides SERS-activity. Spectral collection is provided by a portable Raman analyzer. Providing Chemical Information When & Where You Need It