Determination Of Lysergic Acid Diethylamide Research Articles

Determination of Lysergic Acid Diethylamide and 2-Oxo-3-Hydroxy-LSD in Blood: Validation and Comparison of Two Liquid Chromatography–Tandem Mass Spectrometry Methods

Lysergic acid diethylamide (LSD) is a powerful hallucinogen. Its detection is limited by its low dosage; moreover, LSD is rapidly metabolized into 2-oxo-3-hydroxy-LSD (O-H-LSD). In this study we validated two methods for determination of LSD and O-H-LSD in blood. Method #1 consisted in the upgrade of a previously developed procedure for detection of 163 compounds. Method #2 was specific for LSD and O-H-LSD. Analyses were performed through LC-MS/MS by dynamic (#1) and/or MRM mode (#2), in positive ionization. Transitions were: 324→223,208 m/z for LSD; 356→237,222 m/z for O-H-LSD. Validations were performed following the AAFS’s guidelines. Linearity was good for both methods. Sensitivity was in line with previously validated methods with LOQs at 0.0375 (#1) and 0.025 (#2) ng/mL for LSD and 0.01875 (#1) and 0.0125 (#2) ng/mL for O-H-LSD. Bias and %CV always met the acceptance criteria. RRs were >83%, except for O-H-LSD with method #1. The methods were successfully applied to two real cases. Method #1 proved to be useful for screening purposes, while method #2 can represent a sensitive and reliable tool for confirmation procedures.

The detection and determination of new LSD derivatives in biological material using the LC-MS/MS technique

Psychedelics derived from lysergic acid diethylamide (LSD) represent one of the groups of new psychoactive substances (NPS) that have been gaining popularity in recent years. Due to their very high potency, they are taken in small doses and their identification in biological material is problematic. The compounds from this group, which appear in low concentrations in biological material and metabolise rapidly, require sensitive and selective analytical methods if they are to be detected. A method for the detection and determination of LSD and its new derivatives (ALD-52, 1P-LSD, 1cP-LSD, and 1B-LSD) in whole blood using the LC-MS/MS technique was therefore developed and validated. The analytes were isolated from the blood (0.2 mL) by liquid-liquid extraction with ethyl acetate and acetate buffer (pH 5.5). The method developed was linear in the range of 0.5−10 ng/mL. The limit of detection (LOD) was 0.01 ng/mL for each analyte and the lowest point on the calibration curve was taken as the limit of quantification (LOQ). The method developed is sensitive and samples can be prepared for analysis quickly and easily. The procedure can be applied widely in the analysis of LSD derivatives in biological material –for forensic or clinical purposes, for example –and can easily be expanded for use with further LSD derivatives.

Electrochemistry and analytical determination of lysergic acid diethylamide (LSD) via adsorptive stripping voltammetry

Lysergic acid diethylamide (LSD) is hardly detectable and quantifiable in biological samples because of its low active dose. Although several analytical tests are available, routine analysis of this drug is rarely performed. In this article, we report a simple and accurate method for the determination of LSD, based on adsorptive stripping voltammetry in DMF/tetrabutylammonium perchlorate, with a linear range of 1–90ngL−1 for deposition times of 50s. LOD of 1.4ngL−1 and LOQ of 4.3ngL−1 were found. The method can be also applied to biological samples after a simple extraction with 1-chlorobutane.

THE IDENTIFICATION AND DETERMINATION OF LYSERGIC ACID DIETHYLAMIDE IN NARCOTIC SEIZURES.

Abstract Methods are described by which lysergic acid diethylamide (LSD) can be identified and determined in the presence of heroin, other narcotics and controlled drugs. Thin layer chromatography and an Ehrlich’s-reagent spray is applied to the LSD-containing material. For confirmation, thin layer chromatography after ultra-violet irradiation and acid hydrolysis should be carried out where ergot alkaloids are present. Spectrophotofluorometric analysis of LSD can be used in the presence of heroin, other narcotics and controlled drugs with a standard error of ±2%. If ergot alkaloids are present, thin layer separation, elution and subsequent fluorometric analysis are recommended.

LC-ESI-MS/MS on an ion trap for the determination of LSD, iso-LSD, nor-LSD and 2-oxo-3-hydroxy-LSD in blood, urine and vitreous humor

A method has been developed for the simultaneous determination of lysergic acid diethylamide (LSD), its epimer iso-LSD, and its main metabolites nor-LSD and 2-oxo-3-hydroxy LSD in blood, urine, and, for the first time, vitreous humor samples. The method is based on liquid/liquid extraction and liquid chromatography-multiple mass spectrometry detection in an ion trap mass spectrometer, in positive ion electrospray ionization conditions. Five microliter of sample are injected and analysis time is 12 min. The method is specific, selective and sensitive, and achieves limits of quantification of 20 pg/ml for both LSD and nor-LSD in blood, urine, and vitreous humor. No significant interfering substance or ion suppression was identified for LSD, iso-LSD, and nor-LSD. The interassay reproducibilities for LSD at 20 pg/ml and 2 ng/ml in urine were 8.3 and 5.6%, respectively. Within-run precision using control samples at 20 pg/ml and 2 ng/ml was 6.9 and 3.9%. Mean recoveries of two concentrations spiked into drug free samples were in the range 60-107% in blood, 50-105% in urine, and 65-105% in vitreous humor. The method was successfully applied to the forensic determination of postmortem LSD levels in the biological fluids of a multi drug abuser; for the first time, LSD could be detected in vitreous humor.

Alternative Method for Forensic Determination of Lysergic Acid Diethylamide and Related Compounds in Body Fluids by Liquid?Liquid Extraction and HPLC with Fluorescence Detection

An alternative and practical method is described for simultaneous detection and quantification of the potent hallucinogen lysergic acid diethylamide (LSD) and related compounds in urine and serum samples. The procedure is based on liquid–liquid re-extraction with ethyl acetate and reversed-phase liquid chromatography coupled with fluorescence detection (HPLC–FLD). With detection limits in urine and serum samples of ca 0.07 ng mL−1 for LSD, nor-LSD, and iso-LSD, respectively, the method is well suited to forensic investigations. Application of the method to clinical samples and autopsy material enable selective identification and accurate quantification of LSD and related compounds. Comparison of results with those obtained from an LSD immunoassay (EMIT II) emphasize the need for chromatographic confirmation.

Determination of lysergic acid diethylamide (LSD) in mouse blood by capillary electrophoresis/ fluorescence spectroscopy with sweeping techniques in micellar electrokinetic chromatography.

The separation and on-line concentration of lysergic acid diethylamide (LSD) in mouse blood was achieved by means of capillary electrophoresis/fluorescence spectroscopy using sodium dodecyl sulfate (SDS) as the surfactant. Techniques involving on-line sample concentration, including sweeping micellar electrokinetic chromatography (sweeping-MEKC) and cation-selective exhaustive injection-sweep-micellar electrokinetic chromatography (CSEI-sweep-MEKC) were applied; the optimum on-line concentration and separation conditions were determined. In the analysis of an actual sample, LSD was found in a blood sample from a test mouse (0.1 mg LSD fed to a 20 g mouse; approximately 1/10 to the value of LD(50)). As a result, 120 and 30 ng/mL of LSD was detected at 20 and 60 min, respectively, after ingestion of the doses.

Determination of lysergic acid diethylamide (LSD) by application of online 77 K fluorescence spectroscopy and a sweeping technique in micellar electrokinetic chromatography

The principal advantage of the use of Shopl'skii effect (low temperature spectrum) is that spectral sharpening occurs both in absorption and emission. However, thus far using the technique of capillary electrophoresis/low temperature fluorescence spectroscopy (CE/LTFS) either at 77 or 4.2 K remains difficult to obtain an on-line spectrum, if the analyte is present at low concentration. This paper examines the feasibility of combining the techniques of online concentration and CE/LTFS to identify LSD and related compounds in urine at 77 K. To improve sensitivity, sweeping-micellar electrokinetic chromatography (sweeping-MEKC) and cation-selective exhaustive injection-sweep-micellar electrokinetic chromatography (CSEI-sweep-MEKC) were used for on-line concentration which resulted in detection limits of ∼20 ∼60 ppt, respectively.

Determination of LSD in urine with high-performance liquid chromatography--mass spectrometry.

A rapid, specific, and sensitive high-performance liquid chromatography/mass spectrometry method has been developed for routine determination of lysergic acid diethylamide (LSD) in urine. It includes sample purification by extraction into an organic solvent and back-extraction to an acetate buffer, reversed-phase high-performance liquid chromatography, and detection with a single quadrupole mass spectrometer equipped with an atmospheric pressure ionization electrospray interface. Trideuterated LSD was used as internal standard. The limit of detection was 0.02 ng/mL and the calibration curve was linear from 0.05 to 10 ng/mL. Within-and between-day coefficients of variation were 3.5% and 4.0% respectively and extraction recovery was 91%.

Quantitative determination of LSD and a major metabolite, 2-oxo-3-hydroxy-LSD, in human urine by solid-phase extraction and gas chromatography-tandem mass spectrometry.

An assay has been developed for quantitative determination of lysergic acid diethylamide (LSD) and a major metabolite of LSD in human urine at concentrations as low as 10 pg/mL. In most LSD-positive urine samples the metabolite, 2-oxo-3-hydroxy-LSD, is present at higher concentrations than LSD and can be detected for a longer time than LSD after ingestion of the drug. Urine samples are extracted using Varian Bond Elut Certify extraction cartridges. Confirmatory identification is accomplished by trimethylsilylation of LSD and 2-oxo-3-hydroxy-LSD, followed by gas chromatography-tandem mass spectrometry analysis using positive ion chemical ionization and selected reaction monitoring. Commercially available lysergic acid methylpropylamide and 2-oxo-3-hydroxy-LAMPA are used as internal standards. With selected reaction monitoring, both compounds gave linear calibration curves from 10 pg/mL to 5000 pg/mL. Forty-nine human urine samples that had previously been shown to contain LSD were reanalyzed by the new method. These samples showed an average LSD concentration of 357 pg/mL and an average 2-oxo-3-hydroxy-LSD concentration of 3470 pg/mL. Additional experiments using clinical samples in which two subjects were dosed with LSD support the conclusion that analysis for 2-oxo-3-hydroxy-LSD can permit identification of LSD users for a longer period following ingestion than analysis for the parent drug.

Determination of Lysergic Acid Diethylamide in Body Fluids by High-Performance Liquid Chromatography and Fluorescence Detection—A More Sensitive Method Suitable for Routine Use

A new method for determination of lysergic acid diethylamide (LSD) in body fluids by high-performance liquid chromatography and fluorescence detection was developed based on previously published methods. The new method is suitable for confirmation of samples tested positive by immunoassay, avoiding loss of LSD by absorption to surfaces. The reduced loss of LSD results in improved sensitivity. This is achieved by adding ethylene glycol to the samples, which cover glass surfaces. This principle can similarly be used to improve analysis of other drugs. Body fluids for analysis included urine and whole blood. An internal standard was applied for quantification of LSD. The new method offers satisfying precision data and has a detection limit of less than 0.05 ng/nL.

High-flow ion spray liquid chromatography/mass spectrometry

This paper describes the application of high-flow (up to 2 mL/min) ion spray to the LC/MS determination of compounds of pharmaceutical and environmental interest with standard (HPLC) columns without established postcolumn splitting of the flow. It also illustrates the use of a nonvolatile mobilephase buffer, potassium phosphate, for the LC/MS determination of lysergic acid diethylamide (LSD). To achieve practicality and analytical ruggedness of the system, a liquid shield is placed between the sprayer and the ion sampling capillary, resulting in full-scan low-nanogram determination of quaternary ammonium drugs. The system also allows the use of gradients which is demonstrated for the analysis of carbamate pesticides and in particular for mexacarbate in pond water.

Determination of lysergic acid diethylamide (LSD), iso-LSD, and N-demethyl-LSD in body fluids by gas chromatography/tandem mass spectrometry.

Procedures for detection and quantitation of lysergic acid diethylamide (LSD), iso-LSD, and N-demethyl-LSD by capillary chromatography/tandem mass spectrometry (GC/MS/MS) are presented. Several methods for derivatization, sample introduction, and ionization, in combination with mass spectrometry/mass spectrometry (MS/MS), have been evaluated for overall ionization efficiency and product-ion sensitivity and specificity. Fragmentation pathways derived from low-energy collision-induced dissociation (CID) spectra of protonated LSD, and the protonated trimethylsllyl derivatives of LSD (LSD-TMS) and deuterium-labeled analogs of LSD, have been proposed. Principal dissociations primarily involve the amide and piperidine-ring moieties in which losses of CH3 radical, CH3NH2, CH3NCH2, diethylamine, diethylformamide, and N,N-diethylpropenamide from MH+ are observed. Positive-ion ammonia chemical ionization and subsequent MS/MS analysis of the protonated molecules (MH+) of the trimethylsilyl (TMS) derivatives of LSD, iso-LSD, and N-demethyl-LSD provide a high degree of specificity for identification of these compounds in urine or blood at low-pg/mL concentrations. Negative-ion chemical ionization and GC/MS/MS analysis of the molecular anion (M-) of the trifluoroacetyl (TFA) derivative is well suited for trace-level identification of N-demethyl-LSD, a metabolite of LSD.

Determination of Lysergic Acid Diethylamide (LSD) in Urine by Instrumental High-Performance Thin-Layer Chromatography

An instrumental high-performance thin-layer chromatographic (HPTLC) technique for the determination of lysergic acid diethylamide (LSD) in urine was developed. Before chromatographic separation, a single-step extraction with alkaline wash is performed. The procedure can detect less than 1 microgram LSD/L urine. Results of HPTLC determination are compared with those from a radioimmunoassay (RIA) procedure.

Gas chromatographic-electron-impact mass fragmentometric determination of lysergic acid diethylamide in urine

A sensitive method for the detection and quantitation of lysergic acid diethylamide (LSD) in urine was developed. After initial solvent extraction, the compound was further purified by liquid-liquid extraction or by solid-phase extraction. The trimethylsilyl derivative of LSD was detected by gas chromatography-mass spectrometry (GC-MS) operated in the electron-impact mode with selected-ion monitoring. The presence of LSD was confirmed by comparing retention times and relative abundances of ions of unknowns with that of a standard. The recovery of this procedure was > 89%. The intra-run and inter-run coefficients of variation were 5% and 7%, respectively. This procedure allows detection of LSD concentrations as low as 29 pg/ml. Quantitation of LSD was linear over the concentration range 50–2000 pg/ml.

The determination of LSD in human plasma following oral administration.

A spectrofluorimetric method is described for the assay of lysergic acid diethylamide (LSD) at nanogram levels in human plasma in the presence of a variable blank. The procedure makes use of the observation that ultra-violet light catalyses the hydration of LSD to the non-fluorescent luini-derivative. The difference in fluorescence of plasma extracts before and after intense ultra-violet irradiation is a measure of LSD concentration. This technique has been used for the assay of LSD in plasma following the oral administration of the drug at a dose of approximately 2 μg per kg body weight.

Lysergic Acid Diethylamide: Review and Collaborative Study

Abstract A method for identification and determination of lysergic acid diethylamide (LSD) was studied collaboratively by 11 collaborators on three samples prepared by the Associate Referee. The presumptive test is a valuable tool for rapid preliminary examination. Racemization of the LSD adds further to its identification by paper chromatography. Assay recovery, based on Sandoz LSD-25 reference standard, was 98.0%. Recovery by the same assay, but based on Ergonovine Maleate USP Reference Standard, was 96.5%. The method is recommended for adoption as official, first action.