Determination of tobacco alkaloid enantiomers using reversed phase UPLC/MS/MS

Abstract

Nʹ-Nitrosonornicotine (NNN), a carcinogenic tobacco-specific Nʹ-nitrosamine (TSNA), is on the FDA list of harmful and potentially harmful constituents (HPHCs). Nornicotine, a product of the demethylation of nicotine, is the immediate alkaloid precursor for NNN formation. Nicotine, nornicotine and NNN are optically active. The accumulation of the isomers of nicotine, nornicotine, and NNN impacts their biological activity. In this paper, we report the determination of tobacco alkaloid enantiomers (including nicotine, nornicotine, anabasine, and anatabine) in samples of different tobacco lines using a reversed phase ultra-performance liquid chromatography-tandem mass spectrometer (UPLC/MS/MS) method. Current method demonstates excellent detection capability for all alkaloid enantiomers, with correlation coefficients (r 2 ) > 0.996 within their linear dynamic ranges. The limit of detection (LOD) and limit of quantitation (LOQ) of all analytes are less than 10 ng/mL and 30 ng/mL, respectively. In addition, their recovery and coefficient of variation (CV%) are within 100–115% and 0.2–3.7%, respectively. The method validated in this paper is simple, fast, and sensitive for the quantification of alkaloid enantiomers in tobacco leaf and has been applied to investigations of tobacco alkaloid enantiomer ratios in different tobacco lines and tobacco products.

A Pharmacological Comparison of Two Isomeric Nicotinic Receptor Agonists: The Marine Toxin Isoanatabine and the Tobacco Alkaloid Anatabine

“…An exception was the much lower efficacy of R-isoanatabine at α7 nAChRs ( Figure 3 and Table 2) relative to S-isoanatabine. S-anatabine accounts for approximately 85% of tobacco anatabine [33]. Anabasine also occurs predominantly as the S-form, as does nicotine [33].…”

“…S-anatabine accounts for approximately 85% of tobacco anatabine [33]. Anabasine also occurs predominantly as the S-form, as does nicotine [33].…”

“…Table 1 provides efficacy (current response, Imax) and potency (EC50, the concentration Over a century ago, anatabine was isolated from tobacco leaves, and its structure reported by Spath's laboratory [32]. It is the most abundant minor alkaloid in tobacco, representing about 4% of the alkaloids present in tobacco leaves; nicotine represents >90% of the alkaloids present [33]. Due to it is relative resistance to metabolism, it has become a useful biomarker of tobacco use [34].…”

Pyridine alkaloids with activity in the central nervous system

“… 48 Subsequent studies reported that anatabine exists as an 85:15 scalemic mixture of (−)- and (+)-enantiomers in Nicotiana species. 50 …”

Liquid chromatographic separation of alkaloids in herbal medicines: Current status and perspectives

“…Mobile phase of alkaline pH is applicable for analysis of enantiomers as well. Tobacco alkaloid enantiomers in different tobacco lines were separated in mobile phase with addition of ammonium formate and NH 4 OH [29].…”