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Simultaneous serum nicotine, cotinine, and trans-3'-hydroxycotinine quantitation with minimal sample volume for tobacco exposure status of solid organ transplant patients.

Abstract

Concentrations of nicotine and its metabolites in blood are indicative of patients' current tobacco exposure, and their quantifications have been clinically applied to multiple assessments including demonstration of abstinence prior to heart-lung transplantation. For the purpose of transplant evaluation, the laboratory work up is extensive; thereby an assay with minimal sample volume is preferred. We developed and validated a rapid LC-MS/MS assay to simultaneously quantitate nicotine and its major metabolites, Cotinine and trans-3'-OH-cotinine (3-OH-Cot), in serum. 100μL of serum was spiked with deuterated internal standards and extracted by Oasis HLB solid phase extraction cartridge. Nicotine and metabolites in the reconstituted serum extract were separated by Agilent Eclipse XDB-C8 3.5μm 2.1mm×50mm HPLC column within 4.7min, and quantified by MS/MS with positive mode electrospray ionization and multiple reaction monitoring. Ion suppression was insignificant, and extraction efficiency was 79-110% at 50ng/mL for all compounds. Limit of detection was 1.0ng/mL for nicotine and 3-OH-Cot, and <0.5ng/mL for Cotinine. Linearity ranges for nicotine, cotinine and 3-OH-Cot were 2-100, 2-1000, and 5-1000ng/mL with recoveries of 86-115%. Within-day and twenty-day imprecision at nicotine/cotinine/3-OH-Cot levels of 22/150/90, 37/250/150, and 50/800/500ng/mL were all 1.1-6.5%. The reconstituted serum extracts were stable for at least 7 days stored in the HPLC autosampler at 5°C. Our method correlates well with alternative LC-MS/MS methods. We successfully developed and validated an LC-MS/MS assay to quantitate concentrations of nicotine and its metabolites in serum with minimal sample volume to assess tobacco exposure of heart-lung transplant patients.

Authors+Show Affiliations

Department of Pathology and Genomic Medicine, The Methodist Hospital, Houston, TX 77030, USA. ishu@tmhs.orgNo affiliation info available

Pub Type(s)

Journal Article
Validation Study

Language

eng

PubMed ID

23632003

Citation

Shu, Irene, and Ping Wang. "Simultaneous Serum Nicotine, Cotinine, and Trans-3'-hydroxycotinine Quantitation With Minimal Sample Volume for Tobacco Exposure Status of Solid Organ Transplant Patients." Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, vol. 928, 2013, pp. 139-45.
Shu I, Wang P. Simultaneous serum nicotine, cotinine, and trans-3'-hydroxycotinine quantitation with minimal sample volume for tobacco exposure status of solid organ transplant patients. J Chromatogr B Analyt Technol Biomed Life Sci. 2013;928:139-45.
Shu, I., & Wang, P. (2013). Simultaneous serum nicotine, cotinine, and trans-3'-hydroxycotinine quantitation with minimal sample volume for tobacco exposure status of solid organ transplant patients. Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, 928, 139-45. https://doi.org/10.1016/j.jchromb.2013.03.032
Shu I, Wang P. Simultaneous Serum Nicotine, Cotinine, and Trans-3'-hydroxycotinine Quantitation With Minimal Sample Volume for Tobacco Exposure Status of Solid Organ Transplant Patients. J Chromatogr B Analyt Technol Biomed Life Sci. 2013 Jun 1;928:139-45. PubMed PMID: 23632003.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Simultaneous serum nicotine, cotinine, and trans-3'-hydroxycotinine quantitation with minimal sample volume for tobacco exposure status of solid organ transplant patients. AU - Shu,Irene, AU - Wang,Ping, Y1 - 2013/04/08/ PY - 2012/12/18/received PY - 2013/03/22/revised PY - 2013/03/28/accepted PY - 2013/5/2/entrez PY - 2013/5/2/pubmed PY - 2013/8/21/medline SP - 139 EP - 45 JF - Journal of chromatography. B, Analytical technologies in the biomedical and life sciences JO - J Chromatogr B Analyt Technol Biomed Life Sci VL - 928 N2 - Concentrations of nicotine and its metabolites in blood are indicative of patients' current tobacco exposure, and their quantifications have been clinically applied to multiple assessments including demonstration of abstinence prior to heart-lung transplantation. For the purpose of transplant evaluation, the laboratory work up is extensive; thereby an assay with minimal sample volume is preferred. We developed and validated a rapid LC-MS/MS assay to simultaneously quantitate nicotine and its major metabolites, Cotinine and trans-3'-OH-cotinine (3-OH-Cot), in serum. 100μL of serum was spiked with deuterated internal standards and extracted by Oasis HLB solid phase extraction cartridge. Nicotine and metabolites in the reconstituted serum extract were separated by Agilent Eclipse XDB-C8 3.5μm 2.1mm×50mm HPLC column within 4.7min, and quantified by MS/MS with positive mode electrospray ionization and multiple reaction monitoring. Ion suppression was insignificant, and extraction efficiency was 79-110% at 50ng/mL for all compounds. Limit of detection was 1.0ng/mL for nicotine and 3-OH-Cot, and <0.5ng/mL for Cotinine. Linearity ranges for nicotine, cotinine and 3-OH-Cot were 2-100, 2-1000, and 5-1000ng/mL with recoveries of 86-115%. Within-day and twenty-day imprecision at nicotine/cotinine/3-OH-Cot levels of 22/150/90, 37/250/150, and 50/800/500ng/mL were all 1.1-6.5%. The reconstituted serum extracts were stable for at least 7 days stored in the HPLC autosampler at 5°C. Our method correlates well with alternative LC-MS/MS methods. We successfully developed and validated an LC-MS/MS assay to quantitate concentrations of nicotine and its metabolites in serum with minimal sample volume to assess tobacco exposure of heart-lung transplant patients. SN - 1873-376X UR - https://www.unboundmedicine.com/medline/citation/23632003/Simultaneous_serum_nicotine_cotinine_and_trans_3'_hydroxycotinine_quantitation_with_minimal_sample_volume_for_tobacco_exposure_status_of_solid_organ_transplant_patients_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1570-0232(13)00204-3 DB - PRIME DP - Unbound Medicine ER -