Diagnosis of time of drug exposure from concentrations of original drug and its metabolites

从原药及其代谢物的浓度诊断药物暴露时间

• 批准号: 10670391
• 负责人: KUDO Keiko
• 金额: $ 1.73万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670391/
• 关键词: Time of drug exposure; Triazolam; α -hydroxytriazolam; 4-hydroxytriazolam; Tetracaine; GC-MS; p-butylaminobenzoic acid

In order to diagnose the time of drug exposure from concentrations of original drug and its metabolites in human samples, we first tried to establish the sensitive method to determine original drug and its metabolites simultaneously in triazolam and tetracaine.Triazolam and its major metabolites, α -hydroxytriazolam and 4-hydroxytriazolam were effectively extracted using 3-step solvent extraction procedure followed by tert-butyldimethylsilyl derivatization, and subjected to CG-MS in the negative ion chemical ionization mode. Estazolam was served as internal standard. The calibration curve for each compound was linear in the range from 0.5 to 100 ng/g, and the lower limit of detection was 0.1 ng/g for whole blood and 0.2 ng/g for urine. Tetracaine and its metabolite, p-butylaminobenzoic acid, in the tissues were successfully extracted by a liquid-liquid extraction procedure, and analyzed by CG-MS as free base for tetracaine and as tert-butyldimethylsilyl derivative for the metabolite. Dibucaine and p-dimethylaminobenzoic acid were used as internal standards. The calibration curve for each compound was linear in the range from 10 to 1000 ng/0.5g, and the lower limit of detection was 10ng/g for tetracaine and 0.6 ng/g for the metabolite.A preliminary study was carried out by analyzing concentrations of triazolam and its metabolites in human urine collected every 2 hours from a volunteer who took 1 tablet of triazolam (0.25 mg). α -hydroxytriazolam was found to be excreted to urine faster than 4-hydroxytriazolam, thus the possibility of determining the time of exposure of triazolam by using the concentration ratio of α -hydroxytriazolam to 4-hydroxytriazolam was suggested.

为了从人体样本中原药及其代谢物的浓度来诊断药物暴露时间，我们首先尝试建立同时测定三唑仑和丁卡因中原药及其代谢物的灵敏方法。采用三步溶剂萃取法有效地提取了三唑仑及其主要代谢物α-羟基三唑仑和4-羟基三唑仑 然后进行叔丁基二甲基甲硅烷基衍生化，并在负离子化学电离模式下进行CG-MS。艾司唑仑用作内标。每种化合物的校准曲线在 0.5 至 100 ng/g 范围内呈线性，全血检测下限为 0.1 ng/g，尿液检测下限为 0.2 ng/g。通过液-液萃取程序成功提取了组织中的丁卡因及其代谢物对丁基氨基苯甲酸，并通过 CG-MS 分析丁卡因的游离碱和代谢物的叔丁基二甲基甲硅烷基衍生物。使用地布卡因和对二甲氨基苯甲酸作为内标。每种化合物的校准曲线在 10 至 1000 ng/0.5g 范围内呈线性，丁卡因的检测下限为 10ng/g，代谢物的检测下限为 0.6 ng/g。通过分析每 2 小时从服用 1 片三唑仑的志愿者收集的人体尿液中三唑仑及其代谢物的浓度，进行了初步研究 （0.25 毫克）。发现α-羟基三唑仑比4-羟基三唑仑更快地排泄到尿液中，因此提出利用α-羟基三唑仑与4-羟基三唑仑的浓度比来确定三唑仑暴露时间的可能性。

项目成果

Kudo K et al.: "Simultaneous determination of triazolam and its metabolites in human blood and urine by gas chromatography/mass spectrometry"Legal Med. 1. 159-164 (1999)

Kudo K等人：“通过气相色谱/质谱法同时测定人血液和尿液中的三唑仑及其代谢物”Legal Med。

Kudo K, Ikeda N, Hino Y.: "Simultaneous determination of triazolam and its metabolites in human blood and urine by gas chromatopraphy/mass spectrometry."Legal Med. 1. 159-164 (1999)

Kudo K、Ikeda N、Hino Y.：“通过气相色谱/质谱法同时测定人体血液和尿液中的三唑仑及其代谢物。”Legal Med。

Hino Y, Ikeda N, Kudo K, Tsuji A.: "Sensitive and selective determination of tetracaine and its metabolites in human samples by gas chromatography-mass spectrometry."J Anal Toxicol. (in press.).

Hino Y、Ikeda N、Kudo K、Tsuji A.：“通过气相色谱-质谱法灵敏、选择性地测定人体样品中的丁卡因及其代谢物。”J Anal Toxicol。

Kudo K et al: "Simultaneous determination of triazolam and its metabolites in human blood and urine by gas chromatography/mass spectrometry"Legal Med. 1. 159-164 (1999)

Kudo K等：“气相色谱/质谱法同时测定人血液和尿液中的三唑仑及其代谢物”Legal Med。

Hino Y et al.: "Sensitive and selective determination of tetracaine and its metabolites in human samples by gas chromatography-mass spectrometry."J Anal Toxicol:. (in press).

Hino Y 等人：“通过气相色谱-质谱法灵敏、选择性地测定人体样品中的丁卡因及其代谢物。”J Anal Toxicol：。