Simultaneous determination of nicotine and its nine metabolites in rat blood utilizing microdialysis coupled with UPLC–tandem mass spectrometry for pharmacokinetic application

Abstract

To develop a simple and rapid method for the simultaneous determination of nicotine and its nine metabolites in rat blood, an in vivo microdialysis sampling technique coupled with ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was established for quantitation and characterization of the pharmacokinetics of nicotine and its metabolites. Microdialysis probes were inserted into the jugular vein of Sprague Dawley rats, and dialysates were collected after nicotine (0.5 mg/kg, i.p.) administration. Target analytes and corresponding deuterated internal standards were separated on a hydrophilic interaction liquid chromatography column (HILIC BEH 2.1. × 150 mm, 1.7 μm) and detected by UPLC-MS/MS under multiple reaction monitoring mode. The limits of quantification for nicotine and its nine metabolites ranged from 0.039 to 0.46 ng/mL. Intra- and inter-day precision and accuracy were well within the predefined limits of acceptability (<11 %). Pharmacokinetic results showed that the mean half-lives of nicotine, cotinine, nornicotine, norcotinine, nicotine-N'-oxide, cotinine-N'-oxide, trans-3'-hydroxy-cotinine, nicotine-N-glucuronide, cotinine-N-glucuronide, and trans-3'-hydroxy-cotinine-O-glucuronide in rat plasma were 63, 291, 175, 440, 251, 451, 322, 341, 488, and 516 min, respectively. The blood concentration-time profiles of nicotine and its nine metabolites indicate that nicotine is rapidly consumed after the administration and subsequently cotinine is generated as the main metabolite; meanwhile, cotinine and other eight minor metabolites exhibit longer retention times in rat body.

Simultaneous determination of plasma nicotine and cotinine by UHPLC–MS/MS in C57BL/6 mice and its application in a pharmacokinetic study

“…Methanol failed to obtain good peak shapes simultaneously for nicotine and cotinine. In accordance with four published studies, good peak shapes were simultaneously obtained for both analytes using acetonitrile (containing 0.1% formic acid) and ammonium formate (Abdallah et al, ; Dobrinas et al, ; Loukotkova et al, ; Mao et al, ).…”

“…The quantitative and qualitative ion pairs for both two analytes and IS were similar to those reported in previous methods (Abdallah et al, ; Dobrinas et al, ; Liachenko et al, ; Loukotkova et al, ; Mao et al, ; Miller et al, ; Shakleya & Huestis, ; Stolker et al, ; Yuan et al, ). The 10 μL sample volume was comparable with that in two published methods (10–20 μL) (Kaisar et al, ; Loukotkova et al, ) and much lower than those used in other studies (200–1000 μL) (Baumann et al, ; Dobrinas et al, ; Liachenko et al, ; Miller et al, ; Shakleya & Huestis, ; Stolker et al, ; Yuan et al, ).…”

“…The 10 μL sample volume was comparable with that in two published methods (10–20 μL) (Kaisar et al, ; Loukotkova et al, ) and much lower than those used in other studies (200–1000 μL) (Baumann et al, ; Dobrinas et al, ; Liachenko et al, ; Miller et al, ; Shakleya & Huestis, ; Stolker et al, ; Yuan et al, ). The 1 μL injection volume was lower than those used in all previous studies (2–30 μL) (Abdallah et al, ; Abu‐Awwad et al, ; Baumann et al, ; Dobrinas et al, ; Kaisar et al, ; Loukotkova et al, ; Mao et al, ; Shakleya & Huestis, ; Stolker et al, ; Yuan et al, ). In fact, a 2 μL injection volume will result in significant column overload and appearance of a leading peak.…”

“…Various methods have been developed to determine nicotine and cotinine in plasma, including HPLC, HPLC–MS/MS, and GC–MS (Abdallah, Hammell, Stinchcomb, & Hassan, ; Abu‐Awwad, Arafat, & Schmitz, ; Baumann, Regenthal, Burgos‐Guerrero, Hegerl, & Preiss, ; Dobrinas et al, ; Earla, Ande, McArthur, Kumar, & Kumar, ; Jacob et al, ; Kaisar et al, ; Liachenko, Boulamery, & Simon, ; Loukotkova, VonTungeln, Vanlandingham, & da Costa, ; Mao et al, ; Miller, Norris, Rollins, Tiffany, & Wilkins, ; Onoue, Yamamoto, Seto, & Yamada, ; Shakleya & Huestis, ; Stolker, Niesing, Hogendoorn, Bisoen Rambali, & Vleeming, ; Xu et al, ; Yuan et al, ; Yuan, Kosewick, & Wang, ). However, there are many deficiencies in these published methods, such as the large sample volume requirement (200–1000 μL; Baumann et al, ; Dobrinas et al, ; Liachenko et al, ; Miller et al, ; Shakleya & Huestis, ; Stolker et al, ; Yuan et al, ), complicated and time‐consuming sample pretreatment (Abdallah et al, ; Shakleya & Huestis, ), the long analysis time (5.5–20 min; Abdallah et al, ; Baumann et al, ; Dobrinas et al, ; Loukotkova et al, ; Mao et al, ; Miller et al, ; Onoue et al, ) and poor peak shape (Baumann et al, ). Therefore, this study aims to develop and validate a fast, accurate and robust ultra‐high‐performance liquid chromatography tandem mass/mass spectrometry (UHPLC–MS/MS) method for simultaneous determination of plasma nicotine and cotinine in mice, and apply it to a pharmacokinetic study.…”

Analysis of monofluoroacetic acid in urine by liquid chromatography-triple quadrupole mass spectrometry and preparation of the positive sample by the bioconversion from monofluoroacetamide to monofluoroacetic acid in vitro

“…Ion pair reagent in the mobile phase would present negative effects to MS measurements. Hydrophilic columns were reported to be good at separation of polar compounds without ion pair reagents [16][17][18][19][20][21]. Amide column, which behaves hydrophilic function for chromatographic separation, was introduced to separate FAcOH in urine in this study.…”