Linus Pauling Institute and Department of Pharmaceutical Sciences, Oregon State University, 2900 SW Campus Way, Corvallis, OR, 97331 USA; UIC/NIH Center for Botanical Dietary Supplements Research, University of Illinois at Chicago, 833 S. Wood St., Chicago, IL 60612 USA.
Linus Pauling Institute and Department of Pharmaceutical Sciences, Oregon State University, 2900 SW Campus Way, Corvallis, OR, 97331 USA; UIC/NIH Center for Botanical Dietary Supplements Research, University of Illinois at Chicago, 833 S. Wood St., Chicago, IL 60612 USA.
J Pharm Biomed Anal. 2020 Feb 5;179:112983. doi: 10.1016/j.jpba.2019.112983. Epub 2019 Nov 10.
To evaluate the potential for interactions between botanical dietary supplements and drug metabolism, Phase I clinical pharmacokinetics studies are conducted using an oral cocktail of probe substrates of cytochrome P450 (CYP) enzymes. A sensitive, specific, and fast ultra-high performance liquid chromatography/tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated for determination of caffeine (probe of CYP1A2), tolbutamide (probe of CYP2C9), dextromethorphan (probe of CYP2D6), and alprazolam (probe of CYP3A4/5) in human serum. Stable isotope-labelled analogs were used as internal standards, and sample preparation involved only rapid protein precipitation and centrifugation. The method of standard addition was used for the measurement of caffeine, because commercially available pooled human serum contains caffeine. Out of 18 lots of pooled human serum tested, caffeine was detection in all lots, alprazolam was detected in 13 lots, 8 lots contained dextromethorphan, and no tolbutamide was detected. Only serum prepared from the blood of select individuals was determined to be drug-free. The analytical method was validated with respect to linearity, accuracy and precision, recovery, stability, and matrix effects. The calibration curves were linear over the range of 25-12,000 ng/mL for caffeine, 75-36,000 ng/mL for tolbutamide, 0.05-30 ng/mL for dextromethorphan, and 0.1-60 ng/mL for alprazolam. The intra-assay and inter-assay coefficients of variation (%CV) and %Bias were <13 % (<17 % at the lower limit of quantitation). The recovery of each probe substrate ranged from 84.2%-98.5 %. All analytes were stable during sample storage and handling. Matrix effects were minimized by using stable isotope-labeled internal standards. The method was successfully applied to clinical studies investigating the pharmacokinetic alterations of probe substrates caused by chronic consumption of botanical dietary supplements.
为了评估植物性膳食补充剂与药物代谢之间相互作用的潜力，采用 CYP 酶的口服探针底物混合物进行 I 期临床药代动力学研究。建立并验证了一种灵敏、特异、快速的超高效液相色谱/串联质谱（UHPLC-MS/MS）法，用于测定人血清中的咖啡因（CYP1A2 探针）、甲苯磺丁脲（CYP2C9 探针）、右美沙芬（CYP2D6 探针）和阿普唑仑（CYP3A4/5 探针）。采用稳定同位素标记的类似物作为内标，样品制备仅需快速蛋白沉淀和离心。由于市售混合人血清中含有咖啡因，因此采用标准加入法测定咖啡因。在测试的 18 批混合人血清中，所有批次均检出咖啡因，13 批次检出阿普唑仑，8 批次检出右美沙芬，未检出甲苯磺丁脲。只有从特定个体的血液中制备的血清被确定为无药物。该分析方法在线性、准确度和精密度、回收率、稳定性和基质效应方面进行了验证。咖啡因的校准曲线在 25-12,000ng/mL 范围内呈线性，甲苯磺丁脲在 75-36,000ng/mL 范围内呈线性，右美沙芬在 0.05-30ng/mL 范围内呈线性，阿普唑仑在 0.1-60ng/mL 范围内呈线性。日内和日间变异系数（%CV）和%偏差（%Bias）<13%（定量下限处<17%）。每个探针底物的回收率在 84.2%-98.5%之间。所有分析物在样品储存和处理过程中均稳定。通过使用稳定同位素标记的内标最大限度地减少了基质效应。该方法成功应用于临床研究，研究了慢性食用植物性膳食补充剂对探针底物药代动力学的改变。
