Analysis of drugs in biological matrix
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Methods for analysis of drugs in biological matrix
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Analysis of drugs in biological matrix
- 1. Presented by: Facilitated to: Mr. L. Sanathoiba Singha M. Pharm 2nd Semester Department of Pharmaceutical Analysis. Mrs. Akkamma H. Godihal Assistant Professor Department of Pharmaceutical Analysis. A presentation on Karnataka College of Pharmacy Bengaluru-64, Karnataka. 14/8/2019 1:42 PM ‘Analysis of drugs in biological matrices’
- 2. 4/8/2019 1:42 PM 2 Biological Matrices Urine Blood Hair Saliva Nails Tears Meconium
- 3. 4/8/2019 1:42 PM 3 URINE AND BLOOD: • Urine is the sample of choice for the screening and identification of unknown drugs due to high concentration of drugs in urine. • Improvements in sample preparation, chromatography and in detector techniques have made blood accessible as a screening matrix. • Another great advantage is that drugs can be detected just after intake prior to metabolism and/or filtration. Sample pretreatment: • Conventional sample pretreatment techniques for drugs of abuse analysis in urine samples are liquid–liquid extraction (LLE) solid phase extraction (SPE).
- 4. 4/8/2019 1:42 PM 4 • However, they are rather laborious, time consuming and using large amounts of toxic solvents. Therefore, solventless sample preparation techniques such as Liquid-phase microextraction (LPME) Supercritical fluid extraction (SFE) Solid phase microextraction, had already been proposed. Liquid phase microextraction (LPME) Solid phase microextraction
- 5. 4/8/2019 1:42 PM 5 • In case of urine screening, immunoassays (IA) are used to differentiate between negative and presumably positive samples. • Enzyme-linked immunosorbent assay (ELISA) technique is utilized on whole blood samples for cannabinoids, amphetamines and opiates.
- 6. 4/8/2019 1:42 PM 6 • Capillary electrophoresis (CE) has been proven to have great utility in the analysis and detection of drugs of abuse. But CE is not sensitive enough to be applied to trace analysis due to low injection volumes and limited detection path lengths.
- 7. 4/8/2019 1:42 PM 7 ORAL FLUID SAMPLE (SALIVA): • Detection time of drugs in oral fluid (5–48 hrs) is similar to that in blood (1–2 days) whereas the detection times in urine can be much longer. • Due to short detection times, oral fluid is a feasible matrix for confirmation analysis of driving under the influence of drugs (DUID) cases, where indications of recent drug use is required.
- 8. 4/8/2019 1:42 PM 8 • Samples can be collected with the StatSure SalivaSamplerTM device. Sample pretreatment: Liquid–liquid-extraction (LLE) Solid-phase-extraction (SPE). • Samples were analyzed using gas chromatography–mass spectrometry (GC–MS) with the mass selective detector (MSD) operating in either electron ionization (EI) or negative-ion chemical ionization (NICI) mode. • The compounds analyzed included cannabis, cocaine, amphetamines, opioids, benzodiazepines and other psychoactive medicines.
- 9. 4/8/2019 1:42 PM 9 HAIR SAMPLE: • Drugs are fixed inside the hair matrix, therefore a digestion procedure is necessary before the extraction of drug from the matrix. • An automatic solid-phase extraction method is used. • The method is used along with analysis by gas-chromatography-mass spectrometry (GC/MS) in selected ion monitoring mode (SIM), for the following drugs: codeine, 6-monoacetylmorphine (6-MAM), morphine, cocaine, methadone, ecstasy (MDMA) and Eve (MDE).
- 10. 4/8/2019 1:42 PM 10 NAILS: • Nail samples are usually obtained by cutting the excess overhang of the nail plate using cosmetic nail clippers. • The four key steps in preparing nail samples are - 1. Decontamination; 2. Cutting into small segments; 3. Digestion/Hydrolysis (alkaline, acidic or methanolic) and, 4. Extraction (usually LLE). • Analyzed by GC. • A variety of licit drugs (b-blockers, sedatives, anticoagulant agents, antidepressants and antipsychotics) illicit drugs (cocaine, cannabinoids, morphine and AM related compounds, including their metabolites) has been detected and determined in nails.
- 11. 4/8/2019 1:42 PM 11 TEARS: • Sampling tears is the chief problem to producing precise, reproducible analytical results. • two main procedures for collecting tears are: 1. direct sampling and 2. indirect sampling. Direct sampling comprises collecting tears with capillary tubes and requires previous stimulation. Indirect sampling uses absorbing supports that are very similar to Schrimer strips (classically used to diagnose dry-eye syndrome).
- 12. 4/8/2019 1:42 PM 12 REFERENCES: • Mali, N, Karpe, M, Kadam, V. A review on biological matrices and analytical methods used for determination of drug of abuse .Journal of Applied Pharmaceutical Science. 2011, 01(06). 58-65. • Prabu, S. L, Suriyaprakash, T. N. K. Extraction of Drug from the Biological Matrix: A Review. Applied Biological Engineering - Principles and Practice. 2012, 479-502.
Editor's Notes
- Matrix is the material or tissue in which more specialized structures are embedded.
- Solid-phase extraction (SPE) is a sample preparation process by which compounds that are dissolved or suspended in a liquid mixture are separated from other compounds in the mixture according to their physical and chemical properties
- Liquid-Phase Microextraction. Liquid-Phase Microextraction (LPME) is a methodology that was developed by miniaturising Liquid–Liquid Extraction (LLE), greatly reducing the volume of solvent to just a few microlitres. A supercritical fluid is any substance at a temperature and pressure above its critical point, where distinct liquid and gas phases do not exist. It can effuse through solids like a gas, and dissolve materials like a liquid. Many pressurized gases are actually supercritical fluids. For example, nitrogen has a critical point of 126.2 K (−147 °C) and 3.4 MPa (34 bar). Therefore, nitrogen (or compressed air) in a gas cylinder above this pressure is actually a supercritical fluid. Solid-phase microextraction (SPME) is a technique for separating mixtures of compounds without the use of solvents. SPME uses a fibre coated with a polymer or sorbent extracting phase that extracts chemical compounds from liquid or gas phases.
- Enzyme-linked Immunosorbent Assays (ELISAs) combine the specificity of antibodies with the sensitivity of simple enzyme assays, by using antibodies or antigens coupled to an easily-assayed enzyme.
- Capillary electrophoresis is an analytical technique that separates ions based on their electrophoretic mobility with the use of an applied voltage. The electrophoretic mobility is dependent upon the charge of the molecule, the viscosity, and the atom's radius.
- Electron ionization is an ionization method in which energetic electrons interact with solid or gas phase atoms or molecules to produce ions. Chemical ionization (CI-MS) method uses a reagent gas that is ionized by the electron beam and gives rise to a set of ions that in turn can react with sample. Chemical ionization is a softer technique, resulting in less fragmentation and is often most useful for very fragile compounds fragmenting excessively under EI conditions.
- ecstasy, is a psychoactive drug primarily used as a recreational drug. The desired effects include altered sensations and increased energy, empathy, and pleasure.
- The samples of each person examined are pooled and stored (e.g., in sealed plastic bags) at room temperature with limited light exposure until required for analysis. For surface decontamination, nails are usually washed in an appropriate mixture of reagents (e.g., water, methanol, acetone or surfactant SDS) using an ultrasonic bath.
- The main disadvantages of direct sampling are major dilution of tears induced by stimulation, lack of a standardized time required to collect a sufficient volume of tears and the difficulty of collecting samples from specific sites (e.g., under the eyelid). Dry eye syndrome (DES), also known as keratoconjunctivitis sicca (KCS), is the condition of having dry eyes. Other associated symptoms include irritation, redness, discharge, and easily fatiguedeyes. ... Dry eye occurs when either the eye does not produce enough tears or when the tears evaporate too quickly.