Novel layered nanofiber mat solid-phase extraction coupled with HPLC-MS/MS for determining 15 biomarkers in plasma and urine of rats after exposure to heat-not-burn tobacco products

Abstract

Biomarkers of smoke exposure (BoEs) are often used to assess the safety performance of tobacco products; however, there are few methods available for the simultaneous detection of multiple BoEs. A novel dual-layering packed fibers solid phase extraction (DL-PFSPE) was first developed by stacking two layers of nanofiber mats consisting of polystyrene (PS) nanofibers and polypyrrole (PPy) nanofibers separately, which can simultaneously extract polar and non-polar BoEs from plasma and urine samples. Coupled with HPLC-MS/MS, fifteen BoEs including cotinine, 4-(methyl nitrosamine)-1-(3-pyridyl) butanol (NNAL), N-acetyl-S-(2-carbamoyl ethyl)-L-cysteine (AAMA), N-acetyl-S-(3,4-dihydroxy butyl)-L-cysteine (DHBMA), N-acetyl-S-(3-hydroxypropyl)-L-cysteine (3-HPMA), N-acetyl-S-(3-carboxy-1-methylpropyl)-L-cysteine (3-HMPMA), and phenylmercuric acid (SPMA), 8-isomeric prostaglandin F2α (8-iso-PGF2α), 1-Hydroxy naphthalene (1-OH Nap), 2-hydroxy naphthalene(2-OH Nap), 1-hydroxy phenanthrene (1-OH Phe), 3-hydroxy phenanthrene (3-OH Phe), 2-hydroxy fluorene (2-OH Flu), 9-hydroxy fluorene (9-OH Flu), and 1-hydroxy pyrene (1-OH Pyr) in the plasma and urine of experimental rats exposed to short-term (2 h) smoke exposure can be analyzed. The coefficient of determination (R2) of the working curves for blood and urine samples was greater than 0.99, and the RSDs were 3.4 % (2-OH Nap) to 7.2 % (2-OH Ful) in plasma, 3.3 % (cotinine) to 7.6 % (2-OH Nap) in urine samples. Intra-day and inter-day coefficients of variation were all less than 10 %. The analysis of 15 BoEs in the plasma and urine samples from experimental rats after short-term (2 h) exposure to heat-not-burn tobacco products revealed a significant increase of some BoEs in rats’ bodies. The proposed analytical strategy not only establishes a novel preparation of extraction cartridge but also confirms it as a promising alternative method for the accurate determination of BoEs in plasma and urine samples with good stability and reproducibility.