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Validation of a high-throughput method for simultaneous determination of areca nut and tobacco biomarkers in hair using microwave-assisted extraction and isotope dilution liquid chromatography tandem mass spectrometry.
J Pharm Biomed Anal. 2022 Jul 15; 216:114775.JP

Abstract

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 (r2 > 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.

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Publisher Full Text (DOI)

Authors+Show Affiliations

Chen HC
Institute of Medicine, Chung Shan Medical University, Taichung 40201, Taiwan.
Chen YY
Department of Stomatology, Tungs' Taichung MetroHarbor Hospital, Taichung, Taiwan.
Chao MR
Department of Occupational Safety and Health, Chung Shan Medical University, Taichung 40201, Taiwan.
Chang YZ
Institute of Medicine, Chung Shan Medical University, Taichung 40201, Taiwan; Drug Testing Center, Department of Clinical Laboratory, Chung Shan Medical University Hospital, Taichung 40201, Taiwan. Electronic address: yzc@csmu.edu.tw.

MeSH

AlkaloidsArecaArecolineBiomarkersChromatography, LiquidCotinineHairHumansIsotopesMicrowavesNicotineNutsTandem Mass SpectrometryTobaccoTobacco Products

Pub Type(s)

Journal Article

Language

eng

PubMed ID

35490505

Citation

Chen, Hsiu-Chuan, et al. "Validation of a High-throughput Method for Simultaneous Determination of Areca Nut and Tobacco Biomarkers in Hair Using Microwave-assisted Extraction and Isotope Dilution Liquid Chromatography Tandem Mass Spectrometry." Journal of Pharmaceutical and Biomedical Analysis, vol. 216, 2022, p. 114775.
Chen HC, Chen YY, Chao MR, et al. Validation of a high-throughput method for simultaneous determination of areca nut and tobacco biomarkers in hair using microwave-assisted extraction and isotope dilution liquid chromatography tandem mass spectrometry. J Pharm Biomed Anal. 2022;216:114775.
Chen, H. C., Chen, Y. Y., Chao, M. R., & Chang, Y. Z. (2022). Validation of a high-throughput method for simultaneous determination of areca nut and tobacco biomarkers in hair using microwave-assisted extraction and isotope dilution liquid chromatography tandem mass spectrometry. Journal of Pharmaceutical and Biomedical Analysis, 216, 114775. https://doi.org/10.1016/j.jpba.2022.114775
Chen HC, et al. Validation of a High-throughput Method for Simultaneous Determination of Areca Nut and Tobacco Biomarkers in Hair Using Microwave-assisted Extraction and Isotope Dilution Liquid Chromatography Tandem Mass Spectrometry. J Pharm Biomed Anal. 2022 Jul 15;216:114775. PubMed PMID: 35490505.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Validation of a high-throughput method for simultaneous determination of areca nut and tobacco biomarkers in hair using microwave-assisted extraction and isotope dilution liquid chromatography tandem mass spectrometry. AU - Chen,Hsiu-Chuan, AU - Chen,Ya-Yi, AU - Chao,Mu-Rong, AU - Chang,Yan-Zin, Y1 - 2022/04/18/ PY - 2022/01/31/received PY - 2022/04/10/revised PY - 2022/04/15/accepted PY - 2022/5/2/pubmed PY - 2022/5/31/medline PY - 2022/5/1/entrez KW - Areca alkaloids KW - Hair KW - LC-MS/MS KW - Tobacco alkaloids SP - 114775 EP - 114775 JF - Journal of pharmaceutical and biomedical analysis JO - J Pharm Biomed Anal VL - 216 N2 - 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 (r2 > 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. SN - 1873-264X UR - https://www.unboundmedicine.com/medline/citation/35490505/Validation_of_a_high_throughput_method_for_simultaneous_determination_of_areca_nut_and_tobacco_biomarkers_in_hair_using_microwave_assisted_extraction_and_isotope_dilution_liquid_chromatography_tandem_mass_spectrometry_ DB - PRIME DP - Unbound Medicine ER -