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    涡旋辅助分散液液微萃取-悬浮固化-高效液相色谱法测定酱油、食醋中苯甲酸和山梨酸的含量

    Determination of Benzoic Acid and Sorbic Acid in Soy Sauce and Vinegar by High Performance Liquid Chromatography After Vortex-Assisted Dispersive Liquid-Liquid Microextraction Based on Solidification of Floating Organic Drop

    • 摘要: 取酱油或食醋样品0.25 g置于15 mL聚丙烯离心管中,用水稀释至2.0 mL,加入1 mol·L–1盐酸溶液调节pH至2。用1 mL移液枪吸取250 μL 1-十二醇,快速注入样品溶液中,涡旋萃取60 s,形成乳化浊液,离心5 min。将离心管置于−20 ℃冰箱中5 min后取出,1-十二醇层固化成块状。用不锈钢药匙小心将其取出,置于另一支干净的15 mL离心管中。样品溶液中再加入250 μL 1-十二醇,用相同的方法重复萃取一次。合并两次1-十二醇层,加热融化后,用甲醇稀释至5.0 mL,经0.22 μm有机滤膜过滤,以Agilent Eclipse XDB-C18色谱柱为固定相,以体积比70∶30的20 mmol·L–1乙酸铵溶液-甲醇混合液为流动相,在检测波长230 nm下采用高效液相色谱法测定滤液中苯甲酸和山梨酸的含量,外标法定量。结果显示:苯甲酸和山梨酸的质量浓度在1.0~50 mg·L–1内与对应的峰面积呈线性关系,检出限(3S/N)分别为0.61,0.35 mg·kg–1。对酱油和食醋样品进行3个浓度水平的加标回收试验,苯甲酸和山梨酸的回收率分别为95.6%~102%和95.3%~98.1%,测定值的相对标准偏差(n=6)均小于5.0%。方法用于实际样品分析,苯甲酸和山梨酸均被不同程度检出。

       

      Abstract: The soy sauce or vinegar sample (0.25 g) was placed in a 15 mL-polypropylene centrifuge tube, and dilute to 2.0 mL with water, and 1 mol·L–1 hydrochloric acid solution was added to adjust pH to 2. 250 µL of 1-dodecanol was taken with a 1 mL-pipette gun, and quickly injected into the sample solution. The solution was vortexed for 60 s to form an emulsified turbid solution and centrifuged for 5 min. The centrifuge tube was placed in the refrigerator at −20 ℃ for 5 min and removed, and the 1-dodecanol layer was solidified into a block, which was carefully removed with a stainless steel medicine spoon, and placed in another clean 15 mL-centrifuge tube. 250 µL of 1-dodecanol was added into the sample solution, and the extraction was repeated in the same way. The 1-dodecanol layers were combined, heated, melt, and diluted to 5.0 mL with methanol, and the solution was passed through a 0.22 µm organic filter membrane. Benzoic acid and sorbic acid in the filtrate were determined by high performance liquid chromatography with Agilent Eclipse XDB-C18 column as the stationary phase and a mixture of 20 mmol·L–1 ammonium acetate solution and methanol at a volume ratio of 70∶30 as the mobile phase, using the external standard method at a detection wavelength of 230 nm. As shown by the results, linear relationships between the corresponding peak areas and mass concentrations of benzoic acid and sorbic acid were kept in the range of 1.0–50 mg·L–1, with detection limits (3S/N) of 0.61 mg·kg–1 and 0.35 mg·kg–1, respectively. Test for recovery was made by standard addition method at three concentration levels on soy sauce and vinegar samples, giving results in the range of 95.6%−102% for benzoic acid and 95.3%−98.1% for sorbic acid, respectively, with RSDs (n=6) of the determined values less than 5.0%. This method was used for the analysis of actual samples, and benzoic acid and sorbic acid were detected to varying degrees.

       

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