Extraction from Moist Snuff with Artificial Saliva of Benzo[a]pyrene and Other Polycyclic Aromatic Hydrocarbons

Abstract

SummaryThe present study evaluated in vitro extractability of various polycyclic aromatic hydrocarbons (PAHs) from moist snuff, when the extracting agent was water or artificial saliva. The extraction was performed on nine brands of moist snuff samples that are commercially available and were purchased from the market in January 2018. The moist snuff brands were selected to represent brands with different tobacco cut size descriptors and flavors. For the measurement of PAHs, two different analytical methods were used, an HPLC (High Performance Liquid Chromatography) method for measuring only benzo[a]pyrene (BaP) and a GC/MS/MS (Gas Chromatography Tandem Mass Spectrometry) method for measuring 21 PAHs (including BaP). These methods were modifications of preexistent methods reported in the literature. The results for BaP indicated that by extracting 500 mg of freeze-dried moist snuff with 6 portions of 20 mL water (120 mL), or with 4 portions of 20 mL artificial saliva, followed by two portions of 20 mL water, the BaP remains close to 100% in the solid material and it is not detected in the extracting solution. PAHs with a molecular weight similar or heavier than BaP also showed no extractability. Lighter PAHs such as fluorene, phenanthrene, anthracene, and 5-methylanthracene showed a relatively good extractability. An intermediate group including fluoranthene, pyrene, and benz[a]anthracene showed some extractability in the conditions of this in vitro experiment. This study is not a substitute for clinical studies regarding PAH uptake in human users of moist snuff. However, the results indicate very limited bioavailability of BaP and heavier PAHs from moist snuff. Higher, but variable bioavailability was indicated for lighter PAHs. Important implications of these findings are that: 1) measurably different BaP content of two moist snuff products is unlikely to result in any meaningfully different consumer exposure to BaP; and 2) biomarkers for one PAH cannot necessarily be used as a reliable indicator of exposure to another PAH, particularly if the molecular weights of the precursor PAHs differ since their bioavailabilities can be very different. [Beitr. Tabakforsch. Int. 28 (2019) 214–223]

Extraction of Benzo[a]pyrene from Moist Snuff with Water or Artificial Saliva (Part 2)

“…The R 2 value for the linear calibration was 0.9999. The method of HPLC analysis of BaP in moist snuff samples was very similar to the one previously reported (6) and no additional validation of the HPLC procedure has been made. The extraction was assumed to be 100%, previous work (6) indicating that no detectable BaP is found using a second extraction of the samples.…”

“…The method of HPLC analysis of BaP in moist snuff samples was very similar to the one previously reported (6) and no additional validation of the HPLC procedure has been made. The extraction was assumed to be 100%, previous work (6) indicating that no detectable BaP is found using a second extraction of the samples. However, the good reproducibility (relative standard deviation RSD% = 1.17%) of the measurement of the lowest calibration point (0.078 ng/mL) indicated that this value can be used as the LOQ value.…”

“…The transfer of BaP from moist snuff to a human user is likely to involve an extraction process from the snuff matrix into saliva. In order to better understand this process, an in vitro study was performed and reported (6) to evaluate the extraction of BaP from several brands of moist snuff into water or into artificial saliva. The study (6) established that most BaP remains in the solid part of extracted moist snuff and basically does not go into the solution.…”

“…In order to better understand this process, an in vitro study was performed and reported (6) to evaluate the extraction of BaP from several brands of moist snuff into water or into artificial saliva. The study (6) established that most BaP remains in the solid part of extracted moist snuff and basically does not go into the solution. The resulting level of BaP in the solid part of extracted moist snuff was around 100% of the initial level prior to extraction, but the variation in the results showed remaining levels of BaP in the solid extracted snuff between 95% and 109%, for either the water or artificial saliva extraction.…”

GC-MS/MS Method for Determination of Polycyclic Aromatic Hydrocarbons in Herbal Medicines

“…Analyses of PAHs have been conducted in many areas, such as air [13], soil [14,15], sediment [16], sludgy [17], river and wastewater [18][19][20], smoke products [21,22], cosmetic products [23] and foods [24][25][26][27][28][29]. Recently, various analytical methods of PAHs for herbal medicines characterized by diverse matrices have been reported as follows.…”