烟草香料广泛用于传统烟草产品中,电子尼古丁,加热烟草制品,还有鼻烟.为了抑制高水分含量引起的真菌生长,防腐剂,如苯甲酸(BA),山梨酸(SA),和对羟基苯甲酸酯经常掺入烟草香料中。尽管如此,食用超过安全阈值的防腐剂可能会带来健康风险。因此,这些防腐剂的分析测定对于质量保证和消费者保护至关重要。例如,BA和SA可在易感个体中引起不良反应,包括哮喘,荨麻疹,代谢性酸中毒,和抽搐。对羟基苯甲酸酯,因为它们的内分泌活动,被归类为内分泌干扰化学物质。尽管进行了广泛的研究,同时定量痕量亲水性(BA和SA)和疏水性(对羟基苯甲酸甲酯,对羟基苯甲酸乙酯,对羟基苯甲酸异丙酯,对羟基苯甲酸丙酯,对羟基苯甲酸丁酯,对羟基苯甲酸异丁酯,和对羟基苯甲酸苄酯)防腐剂在烟草香料中仍然具有挑战性。传统的液相萃取与高效液相色谱(HPLC)联用通常会导致高的假阳性率和灵敏度不足。相比之下,串联质谱提供了高灵敏度和特异性;然而,它的广泛应用受到费力的样品制备和巨大的运营成本的限制。因此,建立一种快速、灵敏的烟用香精中9种防腐剂的样品前处理和分析方法至关重要。在这项研究中,同时测定九种防腐剂(SA,基于三相中空纤维液相微萃取(3P-HF-LPME)技术结合HPLC,建立了烟草香精中的BA和7种对羟基苯甲酸酯)。为了获得最佳的预处理条件,萃取溶剂类型,样品相pH值,受体相pH,样品相体积,提取时间,氯化钠的质量分数,进行了检查。此外,HPLC参数,包括紫外检测波长和流动相组成,是精致的。最佳提取条件为:以二己醚为提取溶剂,15mL样品溶液(pH4)用作样品相,氢氧化钠水溶液(pH12)用作受体相,并在800r/min下进行30min的提取。色谱分离是使用AgilentPoroshell120EC-C18柱(100mm×3mm,2.7μm)和包含甲醇的流动相,0.02mol/L乙酸铵水溶液(含0.5%乙酸),和乙腈进行梯度洗脱。在优化条件下,九种目标分析物在各自的线性范围内表现出良好的线性关系,相关系数(r)≥0.9967,检测限(LODs)和定量限(LOQs)分别为0.02-0.07mg/kg和0.08-0.24mg/kg,分别。在两个峰值水平下,9种目标分析物的富集因子(EF)和提取回收率(ERs)分别为30.6-91.1和6.1%-18.2%,分别。9种目标分析物的回收率范围为82.2%至115.7%,相对标准偏差(RSD)(n=5)小于14.5%,中等和高水平。开发的方法很简单,精确,敏感,非常适合于烟用香精样品中防腐剂的快速筛选。
Tobacco flavors are extensively utilized in traditional tobacco products, electronic nicotine, heated tobacco products, and snuff. To inhibit fungal growth arising from high moisture content, preservatives such as benzoic acid (BA), sorbic acid (SA), and parabens are often incorporated into tobacco flavors. Nonetheless, consuming preservatives beyond safety thresholds may pose health risks. Therefore, analytical determination of these preservatives is crucial for both quality assurance and consumer protection. For example, BA and SA can induce adverse reactions in susceptible individuals, including asthma, urticaria, metabolic acidosis, and convulsions. Parabens, because of their endocrine activity, are classified as endocrine-disrupting chemicals. Despite extensive research, the concurrent quantification of trace-level hydrophilic (BA and SA) and hydrophobic (methylparaben, ethylparaben, isopropylparaben, propylparaben, butylparaben, isobutylparaben, and benzylparaben) preservatives in tobacco flavors remains challenging. Traditional liquid phase extraction coupled with high performance liquid chromatography (HPLC) often results in high false positive rates and inadequate sensitivity. In contrast, tandem mass spectrometry offers high sensitivity and specificity; however, its widespread application is limited by laborious sample preparation and significant operational costs. Therefore, it is crucial to establish a fast and sensitive sample pretreatment and analysis method for the nine preservatives in tobacco flavors. In this study, a method for the simultaneous determination of the nine preservatives (SA, BA and seven parabens) in tobacco flavor was established based on three phase-hollow fiber-liquid phase microextraction (3P-HF-LPME) technology combined with HPLC. To obtain the optimal pretreatment conditions, extraction solvent type, sample phase pH, acceptor phase pH, sample phase volume, extraction time, and mass fraction of sodium chloride, were examined. Additionally, the HPLC parameters, including UV detection wavelength and mobile phase composition, were refined. The optimal extraction conditions were as follows: dihexyl ether was used as extraction solvent, 15 mL sample solution (pH 4) was used as sample phase, sodium hydroxide aqueous solution (pH 12) was used as acceptor phase, and the extraction was carried out at 800 r/min for 30 min. Chromatographic separation was accomplished using an Agilent Poroshell 120 EC-C18 column (100 mm×3 mm, 2.7 μm) and a mobile phase comprising methanol, 0.02 mol/L ammonium acetate aqueous solution (containing 0.5% acetic acid), and acetonitrile for gradient elution. Under the optimized conditions, the nine target analytes showed good linear relationships in their respective linear ranges, the correlation coefficients (r) were ≥0.9967, limits of detection (LODs) and quantification (LOQs) were 0.02-0.07 mg/kg and 0.08-0.24 mg/kg, respectively. Under two spiked levels, the enrichment factors (EFs) and extraction recoveries (ERs) of the nine target analytes were 30.6-91.1 and 6.1%-18.2%, respectively. The recoveries of the nine target analytes ranged from 82.2% to 115.7% and the relative standard deviations (RSDs) (n=5) were less than 14.5% at low, medium and high levels. The developed method is straightforward, precise, sensitive, and well-suited for the rapid screening of preservatives in tobacco flavor samples.