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Simultaneous Determination of Sugar Alcohols in Cigarettes by LC-MS/MS

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Simultaneous Determination of Sugar Alcohols in Cigarettes by LC-MS/MS Simultaneous determination of sugar alcohols in cigarettes by LC-MS/MS Tomoaki Ogawa 1 , Tetsuya Tobita 1 , Norimichi Orikata 1 , Shinsuke Sato 2 , Masaki Ninomiya 1 1 Japan Tobacco INC., Product Quality Research Center, 2 JTI Ökolab GmbH Introduction & Objectives Results & Discussion Sugar alcohols are used as sweeteners and humectants in the food industry. Some sugar alcohols are Method validation present in tobacco leaves and tobacco products such as cigarettes. There is a CORESTA recommended The peaks for the nine sugar alcohols in the mixed standard solution were clearly separated (Figure 1). The method for analysis of sorbitol in tobacco and tobacco products 1, and Wang et al. recently reported a calibration curves for the sugar alcohols were linear with good correlation coefficients (R2 > 0.999). The limits method for some sugar alcohols, sugars, and humectants in tobacco products 2. However there is no of detection were approximately 20 µg/g. All sugar alcohols had good average recovery rates (98 %–115 %) 2018_STPOST37_Ogawa.pdf method for simultaneous determination of sugar alcohols which might be contained in cigarette cut filler. The with relative standard deviations between 2.4 % and 13.3 % (Table 5). purpose of this study was to develop a method for simultaneous determination of such nine sugar alcohols Table 5 Calibration range and liniarity, limits of detection, and recoveries for analytes. (meso-erythritol, xylitol, D-pinitol, D-sorbitol, D-mannitol, myo-inositol, maltitol, lactitol, and palatinit) in cut 3 5 Calibration range Spiked level※ Mean recovery RSD Analyte Liniarity R2 filler. To obtain analytical results with high selectivity, we used liquid chromatography tandem mass (μg/mL) (μg/g) (%) (%) spectrometry (LC-MS/MS). 1 4 meso-erythritol 0.005 - 0.5 0.9998 101.4 104 10.6 2 xylitol 0.005 - 0.5 0.9995 99.8 107 7.0 D-pinitol 0.005 - 0.5 0.9996 102.8 102 6.6 7 D-sorbitol 0.005 - 0.5 0.9996 100.0 115 10.1 Materials & Methods 8 6 9 D-mannitol 0.005 - 0.5 0.9994 100.0 109 12.6 Samples myo-inositol 0.005 - 2.5 0.9999 814.4 100 13.3 • CORESTA monitor test piece No. 8 (CM8) Figure 1. LC-MS/MS chromatogram for sugar alcohols maltitol 0.005 - 0.5 0.9997 106.0 101 3.4 at 0.1 µg/mL. 1: meso-erythritol, 2: xylitol, 3: D-pinitol, • University of Kentucky Reference Cigarette 3R4F lactitol 0.005 - 0.5 0.9999 103.4 98 2.4 4: D-sorbitol, 5: D-mannitol, 6: myo-inositol, 7: maltitol, palatinit 0.005 - 0.5 0.9998 100.0 98 3.9 • 24 commercial cigarettes (A–X) 8: lactitol, and 9: palatinit RSD: relative standard deviation, ※Spiked to CM8 Sample preparation Application to cigarette cut filler Ground cut filler (0.5 g) was mixed with 20 mL of ultrapure water and shaken for 60 min. After shaking, the Sugar alcohols in reference (CM8 and 3R4F) and commercial (24 brands) cigarette cut filler samples were water extract was passed through a filter (PTFE, 0.45 μm) and then ultrafiltrated. The ultrafiltrate was diluted analysed by the developed method. Some sugar alcohols were detected in the CM8, 3R4F, and commercial to 100 times the initial volume with a water-acetonitrile mixture (2:8, v/v) and passed through another filter cigarette cut filler samples (Figure 2). (PTFE, 0.2 μm) before LC-MS/MS analysis. μg/g μg/g μg/g Instrumentation The instrument conditions and parameters are shown in Tables 1–4. sample sample sample Table 1 LC conditions. Table 2 Gradient program of the mobile phase. μg/g μg/g μg/g Instrument: Agilent 1290 Infinity Time Flow rate Mobile phase Column: Imtakt Unison UK-Amino HT, (min) (μL/min) A (%) B (%) 2 x 150 mm, 3μm 0 200 10 90 Guard column: Imtakt guard column system for Unison 7 200 10 90 UK-Amino, 2 x 5 mm 35 200 20 80 Column temerature: 40 ˚C 38 200 90 10 sample sample sample Injection volume: 5 μL 40 200 90 10 μg/g μg/g μg/g Flow rate: 0.2 mL/min 40.1 200 10 90 Mobile phase A: 10 mM ammonium acetate in water 46 200 10 90 Mobile phase B: Acetonitorile Table 3 MS conditions. Table 4 MRM transitions and compound dependent MS parameters. sample sample sample Instrument: AB SCIEX TRIPLE QUAD 5500 Precursor ion Product ion Figure 2. Sugar alcohol contents in cut filler samples (CM8, 3R4F, and 24 commercial cigarette brands). Ionization mode: ESI (-) Analyte RT (min) DP (V) CE (V) (m/z) (m/z) Bars show standard deviations for three measurements. Ion spray voltage: -4500 V - meso-erythritol 4.8 [M+CH COO] 181.1 121.0 -50 -9 Ion source temperature: 350 ˚C 3 - Conclusions curtain gas: 30 psi xylitol 6.5 [M+CH3COO] 211.0 59.0 -40 -32 - nitrogen collision gas: 4 psi D-pinitol 7.1 [M+CH3COO] 253.1 193.0 -58 -12 We developed a LC-MS/MS method for simultaneous analysis of nine sugar alcohols (meso-erythritol, xylitol, ion source gas 1: 50 psi - D-sorbitol 9.5 [M-H] 180.9 88.9 -120 -20 D-pinitol, D-sorbitol, D-mannitol, myo-inositol, maltitol, lactitol, and palatinit) in cigarette cut filler. The method ion source gas 2: 80 psi D-mannitol 10.6 [M-H]- 180.9 88.9 -120 -20 has high selectivity, good linearity, and good reproducibility. Some sugar alcohols were detected in the cut myo-inositol 19.0 [M-H]- 178.9 87.0 -138 -21 fillers of CM8, 3R4F, and commercial cigarettes. maltitol 23.0 [M-H]- 343.1 178.9 -130 -19 lactitol 24.5 [M-H]- 343.1 178.9 -130 -19 References palatinit 26.2 [M-H]- 343.1 178.9 -130 -19 1. CORESTA Recommended Methods No. 61:2015 RT: retention time, DP: dissociation energy, CE: collision energy Determination of 1,2-Propylene Glycol, Glycerol and Sorbitol in Tobacco and Tobacco Products by High Performance Liquid Chromatography (HPLC) 2. Wang, L., Cardenas, R. B. & Watson, C. (2017). An isotope dilution ultra high performance liquid chromatography-tandem mass spectrometry method for the simultaneous determination of sugars and humectants in tobacco products. Journal of Chromatography A, 1514, 95-102. Visit www.jt-science.com Congress2018 - Document not peer-reviewed by CORESTA.
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