Definitive Drug and Metabolite Screening in Urine by UPLC-MS-MS Using a Novel Calibration Technique

被引:9
|
作者
Rosano, Thomas G. [1 ,2 ]
Ohouo, Patrice Y. [2 ]
LeQue, John J. [2 ]
Freeto, Scott M. [3 ]
Wood, Michelle [4 ]
机构
[1] Albany Med Ctr Hosp & Coll, Dept Pathol & Lab Med, Forens Toxicol Lab, 43 New Scotland Ave, Albany, NY 12208 USA
[2] Ctr Med Sci, Natl Toxicol Ctr, Clin & Forens Toxicol Lab, 150 New Scotland Ave, Albany, NY 12208 USA
[3] Waters Corp, Cummings Ctr 100, Suite 407N, Beverly, MA USA
[4] Waters Corp, Hlth Sci Org, Altrincham Rd, Wilmslow, Cheshire, England
关键词
TANDEM MASS-SPECTROMETRY; PERFORMANCE LIQUID-CHROMATOGRAPHY; TIME-OF-FLIGHT; LABELED INTERNAL STANDARD; LC-MS/MS; WHOLE-BLOOD; ABUSE; VALIDATION; MATRIX; TOXICOLOGY;
D O I
10.1093/jat/bkw050
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Drug screening is an essential analytical tool for detection of therapeutic, illicit and emerging drug use. Presumptive immunoassay screening is widely used, while initial definitive testing by chromatography-coupled mass spectrometry is hampered due to complex pre-analysis steps, long chromatography time and matrix effects. The aim of this study is to develop and validate a definitive test for rapid and threshold accurate screening of 33 drugs or metabolites (analytes) in urine. Sample preparation in a 96-well plate format involves rapid glucuronidase hydrolysis followed by dilution, filtration and ultra-performance liquid chromatography-MS-MS analysis. Chromatographic separation, on an ACQUITY UPLCA (R) BEH phenyl column is optimized for a 3-min MS-MS ion acquisition. Matrix effect was normalized by an innovative technique called threshold accurate calibration employing an additional analysis with an analyte spike as an internal standard undergoing the same matrix effect as an analyte in a drug-positive donor specimen. Accuracy and precision, at above and below threshold concentrations, were determined by replicate analysis of control urine pools containing 50, 75, 125 and 150% of threshold concentrations. Accuracy and selectivity were further demonstrated by concordant findings in proficiency and confirmatory testing. The study shows the applicability of definitive testing as an alternative to immunoassay screening and demonstrates a new approach to normalization of matrix effect.
引用
收藏
页码:628 / 638
页数:11
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