Methamphetamine In Urine Samples Research Articles

LC-MS Method Development and Optimization for Small Drug Analysis in Urine

Both amphetamine and methamphetamine are considered to be illegal chemicals, and hence, the purchase, possession, and use of these drugs is forbidden in many nations. Within the fields of forensic and clinical toxicology, there has been a recent uptick in the detection and quantification of illicit substances within urine samples. Objective: To detect and quantify both drugs in urine samples utilizing caffeine as an internal standard with an optimized liquid-liquid extraction procedure. Methods: An alternative rapid and efficient method of liquid chromatography – electron spray ionization – Tandem mass spectrometry (LC – ESI – TMS) was developed and optimized. The chromatographic separation was carried out using an isocratic high-performance liquid chromatography (HPLC) system, and the eluent that was applied was a mixture of 20% acetonitrile and 80% buffer with a pH of 2.6 that included 10mM ammonium acetate and 0.1% trifluoroacetic acid. The run duration was 9 minutes, and the detection was accomplished at 210 nm with a flow rate of 1 mL/min utilizing triple quadruple MSMS to validate ionic transitions following direct infusion and fragmentation of analytes. Results: An excellent linearity was seen in the calibration curves of amphetamine and methamphetamine in urine samples across the concentration range of 0-10 mg/L, with a regression coefficient of 0.91 and 0.97, respectively, for each of these substances. Conclusions: More compounds are able to be identified in urine as chromatographic techniques, such as high-performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS), continue to improve in terms of their sensitivity

Analysis of Methamphetamine in Urine by HPLC with Solid-Phase Dispersive Extraction and Solid-Phase Fluorescence Derivatization.

We have developed a fluorescence detection-liquid chromatography (HPLC-FL) method that involves sample pretreatment by solid-phase dispersive extraction (SPDE) and solid-phase fluorescence derivatization for the simple and rapid analysis of methamphetamine (MA) in urine. This method uses a reversed-phase polymeric solid-phase gel to clean up analytes in SPDE, followed by fluorescence derivatization with 9-fluorenylmethyl chloroformate (FMOC) in the solid-phase. The optimal conditions for SPDE and solid-phase fluorescence derivatization were obtained when J-SPEC PEP was used as the solid-phase gel and 0.5 mmol/L FMOC in 50 mmol/L borate buffer solution (pH 10) was used as the fluorescence derivatization reagent. The recovery experiment of MA in urine yielded a clean chromatogram with no interfering peaks, demonstrating the validity of our method; the recoveries were 83.6% when spiked at a low concentration level (100 ng/mL) and 80.7% when spiked at a high concentration level (1000 ng/mL). Compared with the conventional liquid-phase method, the reaction product (FMOC-MA) of solid-phase fluorescence derivatization had higher stability. Reaction rate constants were calculated by changing the temperature conditions, and physicochemical parameters, including activation energy and activation entropy involved in the degradation reaction, were obtained from the Arrhenius plot and analyzed thermodynamically. Taken together, our results suggest that the HPLC-FL method with SPDE and solid-phase fluorescence derivatization for sample pretreatment provides a simple and rapid means of analyzing MA in urine samples.

Determination of Amphetamine and Methamphetamine in Human Hair by Gas Chromatography Mass Spectrometry

This article develops a combined solid phase extraction (SPE) and gas chromatography – mass spectrometry (GC-MS) procedure for determining amphetamine-type stimulants Amphetamine (AM) and Methamphetamine (MA) in human hair. Hair samples were incubated in methanol containing 1% hydrochloric acid in 18 hours and then subjected to SPE. The obtained extracts were evaporated to dryness, derivatized with heptafluorobutyric anhydride (HFBA) at 70 °C for 30 minutes prior to GC–MS analysis. Gas chromatography mass spectrometry was run on HP5-MS column (30 m × 0.25 mm × 0.25 µm) with detector MS 5975C. Experimentally, the proposed method proved sensitive, simple and time-saving, but quite accurate with a low limit of detection (LOD = 0.05ng/mg) and quantitation (LOQ = 0.15ng/mg).
 Keywords:
 SPE, GC – MS, hair samples, amphetamine, methamphetamine.
 References
 [1] Ming-Ren Fuh, Ti-Yu Wu and Tzuen-Yeuan Lin, Determination of amphetamine and methamphetamine in urine by solid phase extraction and ion-pair liquid chromatography–electrospray–tandem mass spectrometry Talanta, 68 (3) (2006), 987-991. https://doi.org/10.1016/j.talanta.2005.06.057[2] Naresh C. Jain, Thomas C. Sneath, and Robert D. Budd, Rapid Gas-Chomatographic Determination of Amphetamine and Methamphetamine in urine, Clinical Chemistry, 20 (11) (1974) 1460-1462. https://doi.org/10.1093/clinchem/20.11.1460.[3] Dong-liang Lin, Rea-Ming Yin, Ray H. Liu, Gas Chromatography-Mass Spectrometry (GC-MS) Analysis of Amphetamine, Methamphetamine, 3,4-Methylenedioxy- amphetamine and 3,4-Methylenedioxymethamphetamine in Human Hair and Hair Sections, Journal of Food and Drug Analysis, 13(2) (2005) 193-200. https://doi.org/10.38212/2224-6614.2526[4] María Jesús Tabernero, Maria Linda Felli, Ana María Bermejo, Marcello Chiarotti, Determination of ketamine and amphetamines in hair by LC/MS/MS, Anal Bioanal Chem, 395(2009), 2547–2557. https://doi.org/10.1007/s00216-009-3163-4.[5] D.V. Doan, D.Q. Huy, N.D. Hue, T.M. Tri, Determination of methamphetamine in urine samples by gas chromatography mass spectrometry combined with solid phase extraction technique, Journal of Science and Technology 47 (6) (2009) 53-58 (in Vietnamese).[6] Rodger L. Foltz, Allison F. Fentiman, Ruth B. Foltz, GC/MS Assays for Abused Drugs in Body Fluids, National Institute on Drug Abuse, Maryland, 1980.[7] AOAC, Appendix F: Guidelines for Standard Method Performance Requirements, AOAC official methods of analysis, Maryland, 2016.[8] Eunyoung Han, Martin P. Paulus, Marc Wittmann, Heesun Chung, Joon myong Song, Hair analysis and self-report of methamphetamine use by methamphetamine dependent individuals, Journal of Chromatography B, 879 (2011) 541–547. https://doi.org/10.1016/j.jchromb.2011.01.002.
 
 

In situ electrosynthesis of a copper-based metal-organic framework as nanosorbent for headspace solid-phase microextraction of methamphetamine in urine with GC-FID analysis.

For the first time, a fiber coating based on copper metal-organic framework was fabricated on an anodized stainless steel wire by an in situ electrosynthesis approach. The fiber was used for the preconcentration and determination of methamphetamine by headspace solid-phase microextraction followed by gas chromatography-flame ionization detection. The electrosynthesis of thefiber coating was performed under a constant potential of - 1.7V by controlling the electrogeneration of OH- in a solution containing sodium nitrate as the probase, 1,2,4,5-benzenetetracarboxylate acid as the ligand and copper nitrate as the cation source. The coating was characterized using field emission scanning electron microscopy, energy-dispersive X-ray spectroscopy, Fourier transform infrared spectroscopy, and thermogravimetric analysis. The effective parameters on the electrosynthesis, extraction, and desorption processes were thoroughly optimized. Under the optimized conditions, metamphetamine (MAP) was quantified over a linear range of 0.90-1000.0ngmL-1 with R2 > 0.997. A limit of detection of 0.1ngmL-1 was achieved, and intra- and inter-day relative standard deviations were found within the range 3.0-4.4% and 2.8-3.9%, respectively. Finally, the method was successfully applied to determination of MAP in urine samples with good recoveries in the range 85.0-102.5%. Graphical abstract Schematic representation of the in-situ electrochemical synthesis of a Cu-based metal-organic framework and its application in a headspace SPME procedure for the measuring methamphetamine in urine samples followed by GC-MS analysis.

Electrospun polyamide/graphene oxide/polypyrrole composite nanofibers: an efficient sorbent for headspace solid phase microextraction of methamphetamine in urine samples followed by GC-MS analysis

A novel polyamide/graphene oxide/polypyrrole nanofiber was fabricated with the aid of the electrospinning technique and applied in headspace solid phase microextraction.

Determination of amphetamines in biological samples using electro enhanced solid-phase microextraction-gas chromatography.

In this work, an ordered mesoporous carbon (OMC)/Nafion coated fiber for solid-phase microextraction (SPME) was prepared and used as the working electrode for electro-enhanced SPME (EE-SPME) of amphetamines. The EE-SPME strategy is primarily based on the electro-migration and complementary charge interaction between fiber coating and ionic compounds. Compared with traditional SPME, EE-SPME exhibited excellent extraction efficiency for amphetamine (AP) and methamphetamine (MA) with an enhancement factor of 7.8 and 12.1, respectively. The present strategy exhibited good linearity for the determination of AP and MA in urine samples in the range of 10-1000ngmL(-1) and 20-1000ngmL(-1), respectively. The detection limits were found to be 1.2ngmL(-1) for AP and 4.8ngmL(-1) for MA. The relative standard deviations were calculated to be 6.2% and 8.5% for AP and MA, respectively. Moreover, the practical application of the proposed method was demonstrated by analyzing the amphetamines in urine and serum samples with satisfactory results.

Rapid extraction and determination of amphetamines in human urine samples using dispersive liquid–liquid microextraction and solidification of floating organic drop followed by high performance liquid chromatography

A novel, rapid, simple and sensitive dispersive liquid–liquid microextraction method based on the solidification of floating organic drop (DLLME-SFO) combined with high-performance liquid chromatography–ultraviolet detection (HPLC–UV) was used to determine amphetamine and methamphetamine in urine samples. The factors affecting the extraction efficiency of DLLME-SFO such as the kind and volume of the extraction and the disperser solvents, effect of concentration of K2CO3 and extraction time were investigated and the optimal extraction conditions were established. Under the optimum conditions (extraction solvent: 30.0μl 1-undecanol; disperser solvent: 300μl acetonitrile; buffer concentration: 2% (w/v) K2CO3 and extraction time: 1min), calibration curves are linear in the range of 10–3000μgl−1 and limit of detections (LODs) are in the range of 2–8μgl−1. The relative standard deviations (RSDs) for 100μgl−1 of amphetamine and methamphetamine in diluted urine are in the range of 6.2–7.8% (n=7). The method was successfully applied for the determination of amphetamine and methamphetamine in the actual urine samples. The relative recoveries of urine samples spiked with amphetamine and methamphetamine are 87.8–113.2%. The obtained results show that DLLME-SFO combined with HPLC–UV is a fast and simple method for the determination of amphetamine and methamphetamine in urine.

Surfactant-assisted dispersive liquid–liquid microextraction followed by high-performance liquid chromatography for determination of amphetamine and methamphetamine in urine samples

A novel surfactant-assisted dispersive liquid–liquid microextraction (SADLLME) based on solidification of floating organic drop (SFO) combined with high-performance liquid chromatography-ultraviolet detection (HPLC-UV) has been proposed for extraction and determination of amphetamine (AM) and methamphetamine (MA) in urine samples. 1-undecanol and sodium dodecyl sulfate (SDS) were used as extracting solvent and disperser, respectively. Face-centered central composite design (FCCCD) was used for optimization of several factors affecting extraction recovery. The optimal conditions were pH = 6.4, volume of extracting solvent V = 31.0 μL, 0.08 CMC as surfactant concentration, and the amount of NaCl for ionic strength was 0.3% (W/V). The limits of detection (LODs, S/N = 3) of the extraction method were 2 for AM and 3 μg L−1 for MA, with the enrichment factors (EFs) of 56 and 48 folds, respectively. The relative standard deviation (RSD %) for n = 3 was below 5.6% and a good linearity (R2 > 0.991) in the range of 10–2000 μg L−1 was observed. The application feasibility of SFO-SADLLME-HPLC-UV in real sample was investigated by analyzing different real samples and satisfactory results were obtained.

Determination of enantiomeric amphetamines as metabolites of illicit amphetamines and selegiline in urine by capillary electrophoresis using modified β-cyclodextrin

The determination of enantiomeric amphetamine and methamphetamine in urine samples is important in order to distinguish use of the prescription drug selegiline (metabolized to R(−)-A and R(−)-MA) from the illicit use of S(+)-A and S(+)-MA. For the analysis of enantiomeric amphetamine (A) and methamphetamine (MA) in biological samples, the optimization of analytical condition was performed by capillary electrophoresis using chiral selectors including β-cyclodextrin, carboxymethyl-β-cyclodextrin and 2-hydroxypropyl-β-cyclodextrin. We have examined the factors to obtain the best chiral resolutions, separation efficiency and sensitivity, and wide concentration linearity. Optimum resolutions were achieved using 100 m M phosphate buffer, pH 2.5, containing 10 m M of carboxymethyl-β-cyclodextrin. This method was applied for the quantitative determination of enantiomeric amphetamine and methamphetamine in urine samples obtained from patients taking illicit amphetamines or from rats and patients taking selegiline. Acceptable quantitative results in terms of resolution, precision, sensitivity and linearity were obtained from the real urine samples containing wide-ranging concentrations of A and MA by using two concentrations of internal standards, α(+)- (1 μg/ml) and β-phenylethylamine (50 μg/ml).

Amphetamine and methamphetamine determination in urine by reversed-phase high-performance liquid chromatography with simultaneous sample clean-up and derivatization with 1,2-naphthoquinone 4-sulphonate on solid-phase cartridges.

A liquid-solid procedure is proposed for sample clean-up and derivatization of amphetamine and methamphetamine in urine samples. The reagent was 1,2-naphthoquinone 4-sulphonate, and a commercial C18 packing cartridge was used. The samples derivatized at room temperature were chromatographed on a 5-microns Hypersil ODS (250 x 4 mm I.D.) with an elution gradient of acetonitrile-water containing propylamine. Under these conditions, the amines were eluted with short retention times. The procedure was used to determine amphetamine, or methamphetamine with its metabolite amphetamine, in spiked urine samples. The detection limit (at a signal-to-noise ratio of 3) for amphetamine (0.1 microgram/ml) was similar to that obtained with liquid-liquid derivatization and to those obtained with immobilized reagents on a polymeric solid support. The detection limit for methamphetamine (0.4 microgram/ml) was higher than with the liquid-liquid procedure because of the lower reactivity on the cartridge. The precision and accuracy of the method were also studied.

Determination of methamphetamine in urine samples with sodium 1,2-naphthoquinone-4-sulphonate

Optimal conditions have been studied for the determination of methamphetamine in urine samples by an extractive-spectrophotometric method with sodium 1,2-naphthoquinone-4-sulphonate (NQS) as reagent. These conditions are: NaHCO3 pH 10, NQS 6.3 × 10−3 mol/l and heating for 5 min at 45°C. The accuracy and precision of the method were tested. The detection limits were 0.2 mg/l in the standard and 0.9 mg/l when 5 ml of urine sample were taken. The standard deviation of blank urine was evaluated from 12 different samples. The relative errors found in the determination of methamphetamine in urine were lower than 10% if the methamphetamine-amphetamine ratio was higher than 4.

A biotin-avidin based screening test for methamphetamine in urine

A biotin—avidin based screening test for methamphetamine in urine samples has been developed. The assay method utilizes the immunoprecipitin reaction between an antibody to methamphetamine and a conjugate prepared by complexing avidin with a biotinyl amphetamine derivative. The rate of the immunoprecipitin reaction is monitored on the Beckman ARRAY® 360 nephelometer. Methamphetamine inhibits the precipitin reaction, and the extent of inhibition allows the quantitation of methamphetamine in the urine samples. Using a cut-off value of 0.7 μg ml −1, the assay correctly predicted 83 of 84 samples (98.8%) confirmed to be positive by GC—MS (>500 ng ml −1). Of 59 GC—MS confirmed negative samples, 46 samples were found to be negative by this method as compared to 34 samples determined with the EMIT® assay. Within-run and between-run relative standard deviations near the cut-off value were less than 4%. Cross-reactivity with amphetamine was <7%.

A screening method for urinary methamphetamine - latex agglutination inhibition reaction test

Methamphetamine in urine samples from abusers was detected by the latex agglutination inhibition reaction test with latex-antibody (Latex-Ab) and latex-methamphetamine (Latex-MA) reagents. Anti-methamphetamine antibody was produced in rabbits by immunization with bovine serum albumin (BSA)-methamphetamine conjugate. Latex particles were coated with antibodies or with rabbit serum albumin (RSA)-methamphetamine conjugate to obtain Latex-Ab and Latex-MA reagents, respectively. The results are read at 4–5 min after mixing the latex reagents. The sensitivity of this method for methamphetamine was 0.4 μg/ml urine. Methamphetamine analogs (methylephedrine, amphetamine, phentermine, methoxyphenamine, ephedrine, β-phenylethylamine, OH-methamphetamine, OH-amphetamine, and OH-ephedrine) all cross-react in varying degrees, while glucosiurea and albuminurea give false positive results in the tests. Though attention must be paid to these effects this simple and rapid test is suitable for the mass screening of urine samples.