Identification of Suvorexant in Blood Using LC–MS-MS: Important Considerations for Matrix Effects and Quantitative Interferences in Targeted Assays

2019 ◽  
Vol 44 (3) ◽  
pp. 245-255 ◽  
Author(s):  
Britni Skillman ◽  
Sarah Kerrigan
Keyword(s):  
Matrix Effects ◽  
Limit Of Detection ◽  
Chemical Properties ◽  
Internal Standard ◽  
Forensic Toxicology ◽  
Calibration Model ◽  
Physico Chemical ◽  
Limit Of Quantitation ◽  
Liquid Chromatography Tandem Mass ◽  
Electrospray Interface

Abstract Suvorexant (Belsomra®) is a novel dual orexin receptor antagonist used for the treatment of insomnia. The prevalence of suvorexant in forensic samples is relatively unknown, which demonstrates the need for robust analytical assays for the detection of this sedative hypnotic in forensic toxicology laboratories. In this study, suvorexant was isolated from whole blood using a simple acidic/neutral liquid–liquid extraction followed by analysis by liquid chromatography tandem mass spectrometry (LC–MS/MS). Matrix effects were evaluated qualitatively and quantitatively using various extraction solvents, proprietary lipid clean-up devices and source conditions. The method was validated in terms of limit of detection, limit of quantitation, precision, bias, calibration model, carryover, matrix effects and drug interferences. Electrospray is a competitive ionization process whereby compounds in the droplet compete for a limited number of charged sites at the surface. As such, it is capacity-limited, and LC–MS-based techniques must be carefully evaluated to ensure that matrix effects or coeluting drugs do not impact quantitative assay performance. In this report, we describe efforts to ameliorate such effects in the absence of an isotopically labeled internal standard. Matrix effects are highly variable and heavily dependent on the physico-chemical properties of the substance. Although there is no universal solution to their resolution, conditions at the electrospray interface can mitigate these issues. Using this approach, the LC–MS/MS assay was fully validated and limits of detection and quantitation of 0.1 and 0.5 ng/mL suvorexant were achieved in blood.

Determination of ketamine and norketamine in hair and evaluation of polydrug use in ketamine abusers using hair analysis in Korea

2020 ◽  
Author(s):  
Jongsook Rhee ◽  
Jihyun Kim ◽  
Moonhee Jang ◽  
Ilchung Shi ◽  
Sangki Lee
Keyword(s):  
Mass Spectrometry ◽  
Matrix Effects ◽  
Limit Of Detection ◽  
Gas Chromatography Mass Spectrometry ◽  
Controlled Drugs ◽  
Hair Samples ◽  
Limit Of Quantitation ◽  
Liquid Chromatography Tandem Mass ◽  
Forensic Service

Abstract This study evaluated hair samples from 28 subjects who had measurable ketamine levels among the samples requested from 2016 to 2017 into Seoul Institute National Forensic Service in Korea. Ketamine in the hair was extracted by using a solution of 1% hydrochloric acid in methanol for 16 h. Extracts were analyzed using gas chromatography mass spectrometry (GC-MS) or liquid chromatography tandem mass spectrometry (LC-MS-MS). LC-MS-MS method was validated by determining the limit of detection (LOD), limit of quantitation (LOQ), linearity, intra- and inter-accuracy, precision, and matrix effects. In 59 ketamine-positive hair or hair segments from 28 ketamine abusers, the ketamine concentration was found to be in the range of 0.011-335.8 ng/mg (mean, 13.6; median, 1.8), and the norketamine concentration was found to be in the range of 0.001-35.7 ng/mg (mean, 7.5; median, 0.44). The ratio of norketamine to ketamine concentration in hair was in the range of 0.01-1.46 (mean, 0.34; median, 0.26). The distribution of ketamine concentration in hair samples was as follows: 0.01-0.1 ng/mg in 11 samples (18.6%), 0.1-5 ng/mg in 33 samples (55.9%), 5-10 ng/mg in 4 samples (6.8%), 10-15 ng/mg in 2 samples (3.4%), 15-20 ng/mg in 4 samples (6.8%), 40-45 ng/mg in 2 samples (3.4%), 45-50 ng/mg in 1 samples 1.7%) and >100 ng/mg in only 2 samples (3.4%). In the hair of ketamine-abusers, 26 of 28 subjects had simultaneously ketamine with detectable levels of other controlled drugs, including MDMA (n=9), MA (n=3), MDMA/MA (n=3), MDMA/PMA (n=3), MDMA/PMA/MA (n=2), cocaine (n=1), and other drugs (n=5, propofol, zolpidem or benzodiazepines). In most of the hair samples were detected ketamine with other controlled drugs: MDMA (60.7%), MA (28.6%), PMA(17.9%), zolpidem (17.9%), and propofol (14.3%) in the frequency of abuse. In conclusion, most of the ketamine-abusers (92.9%) would be polydrug abusers, who were concomitantly abusing other controlled substances.

Novel and Nonroutine Benzodiazepines and Suvorexant by LC–MS-MS

2020 ◽  
Author(s):  
Luke Garcia ◽  
Nicholas B Tiscione ◽  
Dustin Tate Yeatman ◽  
Lauren Richards-Waugh
Keyword(s):  
Quantitative Analysis ◽  
Limit Of Detection ◽  
Sample Volume ◽  
Calibration Model ◽  
Tandem Mass ◽  
Palm Beach County ◽  
Chromatography Tandem Mass Spectrometry ◽  
Target Analytes ◽  
Limit Of Quantitation ◽  
Liquid Chromatography Tandem Mass

ABSTRACT Benzodiazepines are a commonly prescribed class of drugs that have the potential for abuse. The Palm Beach County Sheriff’s Office received drug seizure submissions that included novel and/or nonroutine benzodiazepines of increasing prevalence from 2017 to 2019. This prompted the development of a method of analysis for these compounds in biological specimens. The method tests for 16 novel and nonroutine benzodiazepines and suvorexant in whole blood by liquid chromatography–tandem mass spectrometry (LC–MS-MS). The target analytes included bromazepam, clobazam, clonazolam, clotiazepam, diclazepam, estazolam, etizolam, flualprazolam, flubromazepam, flubromazolam, loprazolam, lormetazepam, phenazepam, prazepam, suvorexant, tetrazepam and triazolam. The method uses 200 µL of sample, protein precipitation and an instrument run-time of 8 min. The limit of detection was either 1 or 5 ng/mL and the limit of quantitation was either 5 or 25 ng/mL depending on the analyte. The method was validated for quantitative analysis for 15 out of the 17 analytes. Flubromazepam and prazepam were validated for qualitative identification only. A quadratic calibration model (r2 > 0.990) with 1/x weighting was used for all analytes for quantitative analysis. The calibration range was either 5–100 or 25–500 ng/mL depending on the analyte. The coefficient of variation of replicate analyses was within 14% and bias was within ±14%. The method provides a sensitive, efficient and robust procedure for the quantitation and/or qualitative identification of select novel and nonroutine benzodiazepines and suvorexant using LC–MS-MS and a sample volume of 200 µL.

scholarly journals Method Development and Validation for Ultra-High Pressure Liquid Chromatography/Tandem Mass Spectrometry Determination of Multiple Prostanoids in Biological Samples

2013 ◽  
Vol 96 (1) ◽  
pp. 67-76 ◽  
Author(s):  
Rui Yu ◽  
Guiqing Zhao ◽  
John W Christman ◽  
Lei Xiao ◽  
Richard B van Breemen
Keyword(s):  
High Pressure ◽  
Method Development ◽  
Matrix Effects ◽  
Selected Reaction Monitoring ◽  
Calibration Model ◽  
Ultra High Pressure ◽  
Limit Of Quantitation ◽  
Liquid Chromatography Tandem Mass ◽  
Negative Electrospray Ionization ◽  
Development And Validation

Abstract Following oxygenation of arachidonic acid by cyclooxygenase to form prostaglandin H2 (PGH2), a variety of prostanoids can be generated with diverse physiologic effects on pain, inflammation, allergy, cardiovascular system, cancer, etc. To facilitate the quantitative analysis of prostanoids in human serum of cell culture, an ultra-high pressure LC (UHPLC)/MS/MS method was developed and validated for the measurement of six eicosanoids belonging to the cyclooxygenase pathway: PGE2, PGD2, 8-iso-PGF2α, PGF2α, 6-keto-PGF1α, and thromboxane B2 (TXB2 ). Selectivity, matrix effects, calibration model, precision, and accuracy (intraday and interday), lower limit of quantitation (LLOQ), recovery, stability, and sample dilution were evaluated. Fast UHPLC separation was carried out in only 0.5 min with isocratic elution, and each prostanoid was measured using negative electrospray ionization MS with collision-induced dissociation and selected reaction monitoring. UHPLC/MS/MS provided high throughput with peak widths of approximately 3 s and an LLOQ of 0.020 ng/mL for PGE2, 0.027 ng/mL for PGD2, 0.152 ng/mL for 8-iso-PGF2α, 0.179 ng/mL for PGF2α and 6-keto-PGF1α, and 0.013 ng/mL for TXB2.

Evaluating Endogenous GHB Variation in Hair with a Synthetic Hair Matrix

2020 ◽  
Vol 44 (4) ◽  
pp. 354-361 ◽  
Author(s):  
Erin W Lloyd ◽  
Jennifer L Thomas ◽  
Christopher C Donnelly ◽  
Madeline A Montgomery ◽  
Roman P Karas ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Human Head ◽  
Calibration Model ◽  
Head Hair ◽  
Drug Concentrations ◽  
Hair Samples ◽  
Limit Of Quantitation ◽  
Hair Matrix ◽  
Liquid Chromatography Tandem Mass ◽  
Multiple Donors

Abstract The variation in drug concentrations in human head hair from 22 donors was measured using a synthetic hair matrix (SMx™ hair). This matrix is being reported for the first time as a calibrator for an endogenous substance. In comparison to authentic hair or melanin, the synthetic hair provided a reliable batch-to-batch source of liquid matrix similar in composition to authentic hair, but without detectable concentrations of endogenous gamma-hydroxybutyric acid (GHB). Using the synthetic matrix for calibrator samples, validation of a liquid chromatography–tandem mass spectrometry (LC-MS/MS) quantitative method for GHB in human head hair was completed. Validation included the evaluation of the following parameters: accuracy, precision, calibration model, carryover, interferences, limit of detection (LOD), limit of quantitation (LOQ) and processed sample stability. The method was valid over a range of 0.4–12 ng/mg, and its LOD and LOQ were both experimentally estimated to be 0.4 ng/mg. After validation, the variation in endogenous GHB concentrations across multiple donors and locations in the vertex posterior region of the human head were evaluated. Results for 11 non-GHB users showed minimal variability (average 3.0% RSD) across the vertex posterior for hair samples taken from three different areas. There was also low variability (average 1.8% RSD) in repeat samples taken from the same location for 11 other non-users. Endogenous GHB concentrations from the LOD/LOQ to 5.60 ng/mg were determined for the 22 donors using the synthetic hair as a calibrator. These results demonstrate the successful application of a synthetic hair matrix in the analysis of GHB in human hair.

UPLC-MS/MS Method for Determination of Khasianine in Mouse Blood: Application for Its Pharmacokinetic Study

2020 ◽  
Vol 16 (6) ◽  
pp. 705-711
Author(s):  
Lianguo Chen ◽  
Qinghua Weng ◽  
Yijing Lin ◽  
Xiaojie Lu ◽  
Zuoquan Zhong ◽  
...  
Keyword(s):  
Lower Limit ◽  
Pharmacokinetic Study ◽  
Oral Absorption ◽  
Matrix Effects ◽  
Limit Of Detection ◽  
Multiple Reaction Monitoring ◽  
Internal Standard ◽  
Quantitative Detection ◽  
Mouse Blood ◽  
Limit Of Quantitation

Background: The aim of this study was to determine the concentrations of khasianine in mouse whole blood sample and its application for the pharmacokinetics by a rapid, selective and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method. Methods: The blood samples were preprocessed by one-step protein precipitation with acetonitrile. The study was performed on an ACQUITY I-Class UPLC system with a UPLC BEH column. Lannaconitine (internal standard, IS) and khasianine were gradient eluted by a mixture of acetonitrile and water with 0.1% formic acid at a flow rate of 0.4 mL/min. The mass spectrometer was equipped with an Electrospray Ionization (ESI) source in positive mode. The quantitative detection was performed in a multiple reaction monitoring modes at transitions m/z 722.4→70.7 for khasianine and m/z 585.3→119.9 for the corresponding IS. Results: The calibration curve was of good linearity ranging from 0.5 to 1000 ng/mL (r > 0.995). The Lower Limit of Detection (LLOD) and Lower Limit of Quantitation (LLOQ) were 0.2 and 0.5 ng/mL, respectively. The inter-day and intra-day precision (RSD%) were both less than 14%, and the accuracy ranged from 86.6% to 108.3%. The matrix effects were between 98.0% and 103.7%, and the average recovery was better than 67.4%. Conclusion: This assay established a sensitive, rapid, selective UPLC-MS/MS method which was successfully used for the pharmacokinetic study of khasianine in mouse blood, and the absolute availability of khasianine was 0.78% which exhibited a poor oral absorption.

scholarly journals Development of a method for the measurement of dehydroepiandrosterone sulphate by liquid chromatography-tandem mass spectrometry

2005 ◽  
Vol 42 (6) ◽  
pp. 468-474 ◽  
Author(s):  
C A Chadwick ◽  
L J Owen ◽  
B G Keevil
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Lower Limit ◽  
Internal Standard ◽  
Tandem Mass ◽  
Dehydroepiandrosterone Sulphate ◽  
Chromatography Tandem Mass Spectrometry ◽  
Limit Of Quantitation ◽  
Liquid Chromatography Tandem Mass

Background: Dehydroepiandrosterone sulphate (DHEAS) is a steroid that is increasingly being recognized as a potential drug of abuse in many countries. This is due to its reputation as a hormone that may be able to retard the ageing process. The measurement of DHEAS is useful in the diagnosis of medical conditions such as congenital adrenal hyperplasia and polycystic ovary syndrome. Thus, a liquid chromatography-tandem mass spectrometry method has been developed to determine DHEAS concentrations in human serum. Method: The chromatography was performed using a WatersTM 2795 Alliance HT LC system coupled to a Mercury Fusion-RP column fitted with a SecurityGuardTM column. Results: DHEAS and the internal standard, deuterated DHEAS, both had a retention time of 1.5 min. The transition determined by the Micromass QuattroTM tandem mass spectrometer for DHEAS was m/z 367.3>96.7 and for the internal standard m/z 369.3>96.6. The method was linear up to 20 µmol/L; the lower limit of detection and the lower limit of quantitation were both 1 µmol/L. The intra- and interassay imprecision were <11% over a concentration range of 1-18 µmol/L for the in-house quality control and <12% for the intra- and interassay imprecision for the Bio-Rad Lyphocheck QC. Conclusion: The measurement of DHEAS by liquid chromatography-tandem mass spectrometry is robust and has a simple sample preparation procedure with a rapid cycle time of only 4 min.

scholarly journals Development and Application of an UHPLC-MS/MS Method for Comparative Pharmacokinetic Study of Eight Major Bioactive Components from Yin Chen Hao Tang in Normal and Acute Liver Injured Rats

2018 ◽  
Vol 2018 ◽  
pp. 1-12 ◽  
Author(s):  
Yun Wang ◽  
Xinrui Xing ◽  
Yan Cao ◽  
Liang Zhao ◽  
Sen Sun ◽  
...  
Keyword(s):  
Lower Limit ◽  
Limit Of Detection ◽  
Injury Model ◽  
Relative Standard ◽  
Limit Of Quantitation ◽  
Liquid Chromatography Tandem Mass ◽  
Bioactive Ingredients ◽  
Aloe Emodin ◽  
Plasma Pharmacokinetics ◽  
Ultrahigh Performance Liquid Chromatography

Yin Chen Hao Tang (YCHT) is one of the most famous hepatoprotective herbal formulas in China, but its pharmacokinetic investigation in model rats has been rarely conducted. In this study, the hepatic injury model was caused by intraperitoneal injections of carbon tetrachloride (CCl4), and YCHT was orally administered to the model and normal rats. An ultrahigh performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) method was established to analyze the plasma pharmacokinetics of eight major bioactive ingredients from YCHT in both the normal and liver injured rats. The calibration curves presented good linearity (r > 0.9981) in the concentration range. The relative standard deviation (RSD%) of inter- and intraday precision was within 9.55%, and the accuracy (RE%) ranged from -10.72% to 2.46%. The extraction recovery, matrix effect, and stability were demonstrated to be within acceptable ranges. The lower limit of detection (LLOD) and lower limit of quantitation (LLOQ) were around 0.1 ng/mL and 0.5 ng/mL, respectively, which were much lower than those in other related researches. Results reveal that there are significant differences in the pharmacokinetics of scoparone, geniposide, rhein, aloe-emodin, physcion, and chrysophanol in hepatic injured rats as compared to those in control except for scopoletin and emodin. Our experimental results provide a meaningful reference for the clinical dosage of YCHT in treating liver disorders, and the improvement of LLOD and LLOQ can also broaden the range of our method’s application, which is very suitable for quantitating these eight compounds with low levels.

scholarly journals Validation of a New Sensitive Method for the Detection and Quantification of R and S-Epimers of Ergot Alkaloids in Canadian Spring Wheat Utilizing Deuterated Lysergic Acid Diethylamide as an Internal Standard

Toxins ◽  
2021 ◽  
Vol 14 (1) ◽  
pp. 22
Author(s):  
Jensen Cherewyk ◽  
Taylor Grusie-Ogilvie ◽  
Barry Blakley ◽  
Ahmad Al-Dissi
Keyword(s):  
Spring Wheat ◽  
Matrix Effects ◽  
Limit Of Detection ◽  
Ergot Alkaloids ◽  
Internal Standard ◽  
Lysergic Acid ◽  
Lysergic Acid Diethylamide ◽  
Relative Standard ◽  
Validation Parameters ◽  
Sensitive Method

Ergot sclerotia effect cereal crops intended for consumption. Ergot alkaloids within ergot sclerotia are assessed to ensure contamination is below safety standards established for human and animal health. Ergot alkaloids exist in two configurations, the R and S-epimers. It is important to quantify both configurations. The objective of this study was to validate a new ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method for quantification of six R and six S-epimers of ergot alkaloids in hard red spring wheat utilizing deuterated lysergic acid diethylamide (LSD-D3) as an internal standard. Validation parameters such as linearity, limit of detection (LOD), limit of quantification (LOQ), matrix effects, recovery and precision were investigated. For the 12 epimers analyzed, low LOD and LOQ values were observed, allowing for the sensitive detection of ergot epimers. Matrix effects ranged between 101–113% in a representative wheat matrix. Recovery was 68.3–119.1% with an inter-day precision of <24% relative standard deviation (RSD). The validation parameters conform with previous studies and exhibit differences between the R and S-epimers which has been rarely documented. This new sensitive method allows for the use of a new internal standard and can be incorporated and applied to research or diagnostic laboratories.

scholarly journals Rapid detection of dimethyl yellow dye in curry by liquid chromatography-electrospray-tandem mass spectrometry

2010 ◽  
Vol 28 (No. 5) ◽  
pp. 427-432 ◽  
Author(s):  
F. Tateo ◽  
M. Bononi ◽  
F. Gallone
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Limit Of Detection ◽  
Tandem Mass ◽  
Mass Spectral ◽  
Limit Of Quantitation ◽  
Liquid Chromatography Tandem Mass ◽  
High Degree ◽  
Positive Ion

An accurate and rapid method, was devised for the identification and quantitation of dimethyl yellow dye in curry, based on liquid chromatography-tandem mass spectrometry interfaced with electrospray. Mass spectral acquisition was done in positive ion mode applying two fragmentation transitions to provide a high degree of selectivity. The extraction system provided a very high recovery (100.0% to 105.8%) and good results were obtained for the limit of detection (5 &mu;g/kg) and limit of quantitation (16 &mu;g/kg). The applicability of the method to identifing and quantifing the unauthorised dimethyl yellow dye in curry was demonstrated.

Quantification and Stereochemical Composition of R-(−) and S-(+)-Clenbuterol Enantiomers in Bovine Urine by Liquid Chromatography–Tandem Mass Spectrometry

2019 ◽  
Vol 44 (3) ◽  
pp. 237-244
Author(s):  
Benjamín Velasco-Bejarano ◽  
Jahir Bautista ◽  
Martha E Rodríguez ◽  
Raquel López-Arellano ◽  
Roberto Arreguín-Espinosa ◽  
...  
Keyword(s):  
Extraction Efficiency ◽  
Limit Of Detection ◽  
Internal Standard ◽  
High Specificity ◽  
Wilcoxon Test ◽  
Adrenergic Agonist ◽  
Limit Of Quantification ◽  
Bovine Urine ◽  
Enantiomeric Analysis ◽  
Liquid Chromatography Tandem Mass

Abstract Clenbuterol (4-amino-α-[(tert-butylamino)methyl]-3,5-dichlorobenzylalcohol) is a β2-adrenergic agonist. The consumption of meat contaminated with clenbuterol can lead to increased heart rate, blood pressure, anxiety, palpitations and skeletal muscle tremors. Several analytical methods have been developed to identify and quantify clenbuterol in different biological matrices. In this report, we have developed a specific and sensitive analytical method for quantifying clenbuterol and performed an in-depth enantiomeric analysis in bovine urine. The method was evaluated in accordance with international guidelines, and we used an isotopically labeled analog as an internal standard. The extraction efficiency for clenbuterol in bovine urine was &gt; 98%, the limit of detection was 0.05 ng/mL and the limit of quantification was 0.10 ng/mL. Our assay showed high specificity, no carryover was observed and the assay was linear in the range 0.10–8.0 ng/mL. Fifteen bovine urine samples were analyzed (containing clenbuterol), and an enantiomeric analysis was performed. The clenbuterol concentration range was 0.10–10.56 ng/mL across these samples. The levorotatory enantiomer was detected at greater concentrations than the dextrorotatory enantiomer, the ratio being 1.7 ± 0.6 (n = 15), and a statistical difference was observed (P &lt; 0.05) using the Wilcoxon test.

Sign in / Sign up

Export Citation Format

Share Document