In 2019, nearly 3000 U.S. residents developed severe lung injury associated with recent use of e-cigarette or vaping products. The Centers for Disease Control and Prevention (CDC) responded to the outbreak, which was formally defined as e-cigarette, or vaping, product use–associated lung injury (EVALI). CDC Laboratory rapidly developed assays to analyze potentially harmful and addictive substances in bronchoalveolar lavage (BAL) fluid collected from EVALI case patients. This report describes the development and validation of a high-throughput isotope dilution HPLC-MS/MS method for measuring two nicotine biomarkers, cotinine (COT) and trans-3ʹ-hydroxycotinine (HCT), in BAL fluid samples. COT and HCT are the major metabolites of nicotine, the addictive alkaloid presents in tobacco products. This method had good specificity and sensitivity. The limit of detection is 0.033 and 0.0165 ng/mL for COT and HCT, respectively, using only 200 µL of sample volume. The within-run and between-run precision was 2-10%. The overall accuracy, calculated from recovery in three different sample matrices spiked at three concentrations, was 94.8% and 93.6% for COT and HCT, respectively. This novel HPLC-MS/MS method was utilized to characterize recent tobacco exposure in EVALI case patients. This method is useful for characterizing tobacco exposure that may be related to acute and chronic lung injury. Keyword: e-cigarette, vaping, EVALI, nicotine, hydroxycotinine, cotinine

中文翻译：

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用于测量电子烟或电子烟产品使用相关肺损伤 (EVALI) 患者支气管肺泡灌洗液中可替宁和反式-3ʹ-羟基可替宁的 HPLC-MS/MS 方法

2019 年，近 3000 名美国居民因近期使用电子烟或电子烟产品而出现严重肺损伤。美国疾病控制与预防中心 (CDC) 对此次疫情做出了回应，该疫情被正式定义为电子烟或电子烟产品使用相关肺损伤 (EVALI)。CDC 实验室迅速开发了检测方法，用于分析从 EVALI 病例患者收集的支气管肺泡灌洗 (BAL) 液中的潜在有害和成瘾物质。本报告描述了高通量同位素稀释 HPLC-MS/MS 方法的开发和验证，该方法用于测量 BAL 液体样品中的两种尼古丁生物标志物：可替宁 (COT) 和反式 3ʹ-羟基可替宁 (HCT)。COT 和 HCT 是尼古丁的主要代谢物，尼古丁是烟草制品中存在的令人上瘾的生物碱。该方法具有良好的特异性和敏感性。仅使用 200 µL 样品体积，COT 和 HCT 的检测限分别为 0.033 和 0.0165 ng/mL。批内和批间精度为 2-10%。根据加标三种浓度的三种不同样品基质的回收率计算，COT 和 HCT 的总体准确度分别为 94.8% 和 93.6%。这种新颖的 HPLC-MS/MS 方法用于表征 EVALI 病例患者近期的烟草暴露情况。该方法可用于表征可能与急性和慢性肺损伤相关的烟草暴露。关键词： 电子烟、vaping、EVALI、尼古丁、羟基可替宁、可替宁