Concheiro Marta, Shakleya Diaa M, Huestis Marilyn A
Chemistry and Drug Metabolism, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, Baltimore, MD 21224, USA.
Forensic Sci Int. 2009 Jul 1;188(1-3):144-51. doi: 10.1016/j.forsciint.2009.04.005. Epub 2009 Apr 29.
A LCMS method was developed and validated for the simultaneous determination of buprenorphine (BUP), norbuprenorphine (NBUP), buprenorphine-glucuronide (BUP-Gluc) and norbuprenorphine-glucuronide (NBUP-Gluc) in human umbilical cord. Quantification was achieved by selected ion monitoring of precursor ions m/z 468.4 for BUP; 414.3 for NBUP; 644.4 for BUP-Gluc and 590 for NBUP-Gluc. BUP and NBUP were identified by MS(2), with m/z 396, 414 and 426 for BUP, and m/z 340, 364 and 382 for NBUP. Glucuronide conjugates were identified by MS(3) with m/z 396 and 414 for BUP-Gluc and m/z 340 and 382 for NBUP-Gluc. The assay was linear 1-50 ng/g. Intra-day, inter-day and total assay imprecision (%RSD) were <14.5%, and analytical recovery ranged from 94.1% to 112.3% for all analytes. Extraction efficiencies were >66.3%, and process efficiency >73.4%. Matrix effect ranged, in absolute value, from 3.7% to 7.4% (CV<21.8%, n=8). The method was selective with no endogenous or exogenous interferences from 41 compounds evaluated. Sensitivity was high with limits of detection of 0.8 ng/g. In order to prove method applicability, an authentic umbilical cord obtained from an opioid-dependent pregnant woman receiving BUP pharmacotherapy was analyzed. Interestingly, BUP was not detected but concentrations of the other metabolites were NBUP-Gluc 13.4 ng/g, BUP-Gluc 3.5 ng/g and NBUP 1.2 ng/g.
开发并验证了一种液相色谱-质谱联用（LCMS）方法，用于同时测定人脐带中丁丙诺啡（BUP）、去甲丁丙诺啡（NBUP）、丁丙诺啡葡萄糖醛酸苷（BUP-Gluc）和去甲丁丙诺啡葡萄糖醛酸苷（NBUP-Gluc）。通过对BUP的前体离子m/z 468.4、NBUP的m/z 414.3、BUP-Gluc的m/z 644.4和NBUP-Gluc的m/z 590进行选择离子监测来实现定量。BUP和NBUP通过MS(2)进行鉴定，BUP的m/z为396、414和426，NBUP的m/z为340、364和382。葡萄糖醛酸苷结合物通过MS(3)进行鉴定，BUP-Gluc的m/z为396和414，NBUP-Gluc的m/z为340和382。该测定法在1-50 ng/g范围内呈线性。日内、日间和总测定不精密度（%RSD）<14.5%，所有分析物的分析回收率在94.1%至112.3%之间。提取效率>66.3%，过程效率>73.4%。基质效应的绝对值范围为3.7%至7.4%（CV<21.8%，n=8）。该方法具有选择性，在所评估的41种化合物中没有内源性或外源性干扰。灵敏度很高，检测限为0.8 ng/g。为了证明该方法的适用性，对从接受BUP药物治疗的阿片类药物依赖孕妇获得的一根真实脐带进行了分析。有趣的是，未检测到BUP，但其他代谢物的浓度分别为NBUP-Gluc 13.4 ng/g、BUP-Gluc 3.5 ng/g和NBUP 1.2 ng/g。
