Cherstniakova Svetlana A, Garcia Gregory E, Strong John, Bi Daoqin, Weitz Julie, Roy Michael J, Cantilena Louis R
Division of Clinical Pharmacology and Medical Toxicology, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD 20814, USA.
J Anal Toxicol. 2006 Jan-Feb;30(1):21-6. doi: 10.1093/jat/30.1.21.
A rapid and highly sensitive gas chromatography-mass spectrometry (GC-MS) method for simultaneous determination of N,N-diethyl-m-toluamide (DEET) and permethrin with (2)H(10)-phenanthrene (98 atom %) as an internal standard and a separate external standard high-performance liquid chromatography (HPLC) method for pyridostigmine bromide (PB) determination in human plasma were developed and validated. The GC-MS method for DEET and permethrin quantification utilizes a one-step extraction with tert-butylmethylether. The HPLC method for PB quantification involves a solid-phase extraction and UV detection. The range of the analytical method for DEET and permethrin was 1 ng/mL to 100 ng/mL and for PB was 5 ng/mL to 100 ng/mL. Recovery from plasma proved to be more than 80%. The intraday precision ranged from 1.3% to 8% for DEET, from 2.1% to 11.4% for permethrin, and from 3.0% to 4.8% for PB. The interday precision was 3% for DEET, ranged from 5% to 9% for permethrin, and from 5% to 9% for PB. The accuracy for the limit of quantification was 92% +/- 8% relative standard deviation (RSD) for DEET, 112% +/- 11% RSD for permethrin, and 109% +/- 5% RSD for PB. All 3 compounds were stable in human plasma at -80 degrees C for at least 12 months and after 2 freeze-thaw cycles with RSD values ranging from 7.1% (DEET, 80 ng/mL) to 8.1% (DEET, 8 ng/mL), from 2.3% (permethrin, 80 ng/mL) to 11.6 % (permethrin, 8 ng/mL), and from 0.2% (PB, 80 ng/mL) to 3.6% (PB, 8 ng/mL). Both methods were successfully applied to pharmacokinetic/ pharmacodynamic studies of combined exposure of DEET (skin application), permethrin (treated uniforms), and PB (30 mg orally three times/day for four doses) in healthy volunteers (n = 81).
建立并验证了一种快速且高灵敏度的气相色谱 - 质谱联用（GC - MS）方法，用于同时测定N，N - 二乙基 - m - 甲苯胺（避蚊胺，DEET）和氯菊酯，以内标物（2）H（10）-菲（98原子％）进行定量，并建立了一种单独的外标高效液相色谱（HPLC）方法用于测定人血浆中的溴吡斯的明（PB）。用于DEET和氯菊酯定量的GC - MS方法采用叔丁基甲基醚一步萃取。用于PB定量的HPLC方法包括固相萃取和紫外检测。DEET和氯菊酯分析方法的线性范围为1 ng/mL至100 ng/mL，PB的线性范围为5 ng/mL至100 ng/mL。血浆回收率超过80％。日内精密度方面，DEET为1.3％至8％，氯菊酯为2.1％至11.4％，PB为3.0％至4.8％。日间精密度方面，DEET为3％，氯菊酯为5％至9％，PB为5％至9％。定量限的准确度方面，DEET的相对标准偏差（RSD）为92％±8％，氯菊酯为112％±11％，PB为109％±5％。所有3种化合物在 - 80℃的人血浆中至少稳定12个月，经过2次冻融循环后，RSD值范围为7.1％（DEET，80 ng/mL）至8.1％（DEET，8 ng/mL），2.3％（氯菊酯，80 ng/mL）至11.6％（氯菊酯，8 ng/mL），以及0.2％（PB，80 ng/mL）至3.6％（PB，8 ng/mL）。这两种方法均成功应用于健康志愿者（n = 81）中DEET（皮肤涂抹）、氯菊酯（处理过的制服）和PB（口服30 mg，每日3次，共4剂）联合暴露的药代动力学/药效学研究。
