Chen Hsiu-Chuan, Chen Ya-Yi, Chao Mu-Rong, Chang Yan-Zin
Institute of Medicine, Chung Shan Medical University, Taichung 40201, Taiwan.
Department of Stomatology, Tungs' Taichung MetroHarbor Hospital, Taichung, Taiwan.
J Pharm Biomed Anal. 2022 Jul 15;216:114775. doi: 10.1016/j.jpba.2022.114775. Epub 2022 Apr 18.
For people with habits of chewing betel nuts and smoking, the probability of suffering from oral cancer is ten to a hundred times higher than others. Due to the serious health consequences of areca nut and tobacco, a reliable cessation program is needed. Hair is the best option to document long-term exposure. Unfortunately, the research on betel nut in hair did not attract much attention. In this study, a high-throughput method based on microwave-assisted extraction (MAE) and isotope dilution liquid chromatography tandem mass spectrometry (LC-MS/MS) was developed to measure the four biomarkers of betel nuts and cigarettes, including areca alkaloids (arecoline), tobacco alkaloids (nicotine), and their metabolites (arecaidine and cotinine). The hair sample was washed, cut, weighed, and incubated for 3 min MAE with methanol/trifluoroacetic acid, then evaporated and reconstituted for LC-MS/MS analysis. The total experiment time was 50 min. The lower limits of quantification (LOQ) were 5-10 pg/mg. The intra-day and inter-day precision were 2.2-7.6%. Intra-day and inter-day accuracy were - 6.1-8.2%. The method showed good linearity (r > 0.995) over LOQ - 1000 pg/mg concentration ranges. It was successfully applied to analyze 11 subjects of regular areca nut chewers, also smokers. Eight samples were black hair; three samples were naturally black hair with partially gray hair. Measured concentrations in black hair were in the range 56.9 pg/mg to 3.2 ng/mg for arecoline, 12.8 pg/mg to 222.2 pg/mg for arecaidine, 3.8 ng/mg to 33.4 ng/mg for nicotine and 1.1 ng/mg to 6.1 ng/mg for cotinine. The results showed lower levels in gray hair. This method was utilized successfully to analyze pg/mg levels of arecoline, arecaidine, nicotine, and cotinine, and good recoveries were obtained. The mean concentration of arecaidine and cotinine in hair was 15% and 20% of arecoline and nicotine, respectively. A good positive correlation was found between the concentrations of these compounds and self-report. This method improved extraction speed, concentration, and analysis of samples and is useful for monitoring betel nut and smoking cessation programs.
对于有嚼槟榔和吸烟习惯的人来说，患口腔癌的概率比其他人高10到100倍。由于槟榔和烟草对健康有严重影响，因此需要一个可靠的戒烟计划。头发是记录长期接触情况的最佳选择。不幸的是，关于头发中槟榔的研究并未引起太多关注。在本研究中，开发了一种基于微波辅助萃取（MAE）和同位素稀释液相色谱串联质谱（LC-MS/MS）的高通量方法，用于测定槟榔和香烟的四种生物标志物，包括槟榔生物碱（槟榔碱）、烟草生物碱（尼古丁）及其代谢物（去甲槟榔碱和可替宁）。头发样本经过清洗、切割、称重，然后用甲醇/三氟乙酸进行3分钟的微波辅助萃取孵育，接着蒸发并复溶用于LC-MS/MS分析。整个实验时间为50分钟。定量下限（LOQ）为5-10 pg/mg。日内和日间精密度为2.2-7.6%。日内和日间准确度为-6.1-8.2%。该方法在LOQ-1000 pg/mg浓度范围内显示出良好的线性（r>0.995）。它成功应用于分析11名经常嚼槟榔且吸烟的受试者。8个样本为黑发；3个样本为自然黑发且部分头发花白。黑发中测得的槟榔碱浓度范围为56.9 pg/mg至3.2 ng/mg，去甲槟榔碱浓度范围为12.8 pg/mg至222.2 pg/mg，尼古丁浓度范围为3.8 ng/mg至33.4 ng/mg，可替宁浓度范围为1.1 ng/mg至6.1 ng/mg。结果显示白发中的含量较低。该方法成功用于分析槟榔碱、去甲槟榔碱、尼古丁和可替宁的pg/mg水平，并获得了良好的回收率。头发中去甲槟榔碱和可替宁的平均浓度分别为槟榔碱和尼古丁的15%和20%。这些化合物的浓度与自我报告之间存在良好的正相关。该方法提高了样品的萃取速度、浓缩和分析效率，有助于监测槟榔咀嚼和戒烟计划。
