Quantitative Analysis of 26 Opioids, Cocaine, and Their Metabolites in Human Blood by Ultra Performance Liquid Chromatography―Tandem Mass Spectrometry

• Published in: Therapeutic drug monitoring Vol. 35; no. 4; pp. 510 - 521
• Main Authors: RAMIREZ FERNANDEZ, María del Mar, WILLE, Sarah M. R, KUMMER, Nathalie, DI FAZIO, Vincent, RUYSSINCKX, Evi, SAMYN, Nele
• Format: Journal Article
• Language: English
• Published: Hagerstown, MD Lippincott Williams & Wilkins 01-08-2013; Lippincott Williams & Wilkins Ovid Technologies
• Subjects: Analgesics, Opioid - blood; Analgesics, Opioid - metabolism; Biological and medical sciences; Blood; Chromatography; Chromatography, High Pressure Liquid - methods; Cocaine; Cocaine - blood; Cocaine - metabolism; Humans; Mass spectrometry; Medical sciences; Metabolites; Pharmacology. Drug treatments; Plasma; Sensitivity and Specificity; Substance Abuse Detection - methods; Substance-Related Disorders - blood; Tandem Mass Spectrometry - methods; Performance evaluation; Human; Biological fluid; CNS stimulant; Psychotropic; Metabolite; Opiates; Liquid chromatography; Blood; Ester; Mass spectrometry MS/MS; UPLC-MS/MS; Drug of abuse; Cocaine; Quantitative analysis; opioids
• ISSN: 0163-4356; 1536-3694
• DOI: 10.1097/FTD.0b013e31828e7e6b

A sensitive and selective ultra performance liquid chromatographic-tandem mass spectrometric method was developed and fully validated for the simultaneous determination of (in order of chromatographic elution) methylecgonine, pholcodine, morphine, hydromorphone, oxymorphone, norcodeine, codeine, dihydrocodeine, oxycodone, 6-Monoacetylmorphine (6-MAM), hydrocodone, ethylmorphine, norfentanyl, benzoylecgonine, tramadol, normeperidine, meperidine, cocaine, pentazocine, cocaethylene, fentanyl, norbuprenorphine, 2-ethylidine-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), buprenorphine, propoxyphene, and methadone in blood. The matrixes analyzed during the validation experiments were as follows: citrated blank plasma for calibrators, fluoride blank plasma for internal quality control (QC), lyophilized serum for external QC, fluoride plasma and whole blood for authentic samples, and lyophilized serum and whole blood for proficiency testing schemes. Samples were extracted with cation exchange solid-phase extraction cartridges. The target drugs were separated and quantified in a chromatographic run of 8.1 minutes using 0.1% formic acid in water and methanol (with 0.1% formic acid) as mobile phase. The limit of quantification ranged from 0.5 to 2.5 ng/mL depending on the compound and the therapeutic concentration. The intra- and interassay precision was less than 15% for all the compounds (except for pentazocine and EDDP, which was <20%) determined with 2 internal and 2 external QC samples, and the bias was within ±15% (except for methylecgonine, which was <20%). Extraction efficiency was greater than 70% for all the compounds except for EDDP. Matrix effects were evaluated with authentic blood samples (n = 10), and they ranged from 47 to 95%, but they were compensated for most analytes using deuterated analogs as internal standards. Prepared samples were stable for 62 hours in the autosampler. This method was successfully applied to authentic samples (n = 120), involving the use of heroin, cocaine, tramadol, and methadone, and to proficiency testing schemes.