Gheddar L, Raul J S, Kintz P
Institut de médecine légale, laboratoire de toxicologie, Strasbourg, France.
Drug Test Anal. 2019 Jan;11(1):157-161. doi: 10.1002/dta.2445. Epub 2018 Jul 17.
An athlete contested an adverse analytical finding involving hydrochlorothiazide, and requested our laboratory for testing his hair. As there is no reference in the literature about identification of hydrochlorothiazide in hair, several volunteers were first enrolled (4 after a single 25 mg administration and 10 with daily therapeutic treatment). A specific method was developed by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS. Hair samples were decontaminated with dichloromethane and 30 mg were incubated in buffer at pH 7.0 for 15 hours at 50°C. Then, 5 mL ethyl acetate was added for extraction. Linearity was observed for hydrochlorothiazide concentrations ranging from 5 to 2000 pg/mg. The limit of quantification was 5 pg/mg. The coefficients of variation (CVs) of repeatability and matrix effect were lower than 20%. Analysis of the 0-2-cm segment of the 4 volunteers having received a single dose, collected 1 month after administration, was negative at the limit of quantification. The hair of the 10 patients (proximal 2 cm) on daily treatment was positive with concentrations ranging from 12 to 1845 pg/mg, with no correlation between daily dose and concentration. The athlete's hair tested positive for hydrochlorothiazide at 36 pg/mg in the segment corresponding to the period of the urinary control. Since a single exposure to hydrochlorothiazide is not detectable in hair and based on the results of the patients on daily treatment, the concentration found in the athlete has been interpreted as corresponding to repeated exposures, where it was not possible to establish the dosage and the frequency.
一名运动员对一项涉及氢氯噻嗪的不利分析结果提出异议，并请求我们实验室对其头发进行检测。由于文献中没有关于头发中氢氯噻嗪鉴定的参考资料，首先招募了几名志愿者（单次服用25毫克后4名，接受每日治疗的10名）。通过超高效液相色谱 - 串联质谱法（UPLC-MS/MS）开发了一种特定方法。头发样品用二氯甲烷去污，30毫克在pH 7.0的缓冲液中于50°C孵育15小时。然后加入5毫升乙酸乙酯进行萃取。观察到氢氯噻嗪浓度在5至2000 pg/mg范围内呈线性。定量限为5 pg/mg。重复性和基质效应的变异系数（CVs）低于20%。对单次给药后1个月收集的4名志愿者0 - 2厘米段的分析，在定量限处为阴性。10名接受每日治疗患者的头发（近端2厘米）呈阳性，浓度范围为12至1845 pg/mg，每日剂量与浓度之间无相关性。在与尿液检测期相对应的段中，该运动员的头发氢氯噻嗪检测呈阳性，浓度为36 pg/mg。由于在头发中无法检测到单次接触氢氯噻嗪，并且基于每日治疗患者的结果，在该运动员身上发现的浓度被解释为对应于重复接触，在此无法确定剂量和频率。
