同时测定酱油中4种甜味剂LC-MS/MS法的建立

Establishment of LC-MS/MS Method for Simultaneous Determination of Four Sweeteners in Soy Sauce

建立了液质联用同时测定酱油中甜菊糖苷、甜菊双糖苷、甘草酸、甘草次酸的方法。方法经提取溶剂和提取次数优化后,采用液质联用进行检测。试验采用ZORBAX Eclipse Plus C184.6 mm×150 mm 3.5μm为液相色谱柱,以甲醇、0.1%甲酸为流动相,流速为0.4 mL/min,质谱中正负模式同时扫描。试验表明:甜菊糖苷、甜菊双糖苷为正模式扫描;甘草酸、甘草次酸为负模式扫描;提取溶剂为甲醇,提取次数为2次;甜菊糖苷、甜菊双糖苷、甘草酸、甘草次酸在0~40.0 ng/mL范围内r值均为0.999;甜菊糖苷、甜菊双糖苷、甘草酸、甘草次酸在50.0,200和400μg/kg回收率分别在90.5%~94.9%,87.7%~94.6%,86.9%~97.3%和87.1%~96.5%之间;甜菊糖苷、甜菊双糖苷、甘草酸、甘草次酸精密度(n=6)分别为2.21%,2.00%,1.88%和2.72%。该方法操作方便、定性能力强、灵敏度高、准确度高,适合酱油中甜菊糖苷、甜菊双糖苷、甘草酸、甘草次酸的检测。

A method for simultaneous determination of stevioside,steviolbioside,glycyrrhizic acid and glycyrrhetinic acid in soy sauce by LC-MS/MS was established.LC-MS was used for detection after optimizing the extraction solvent and extraction times.Zorbax eclipse plus C184.6 mm×150 mm 3.5μm was used as liquid chromatographic column,while methanol and 0.1%formic acid were used as mobile phase,with flow rate at 0.4 mL/min.Simultaneous scanning of positive and negative modes was in mass spectrometry.The results showed that stevioside and steviolbioside were positive mode scanning;Glycyrrhizic acid and glycyrrhetinic acid were negative mode scanning.The extraction solvent was methanol and 2 times extraction.The r values of stevioside,steviolbioside,glycyrrhizic acid and glycyrrhetinic acid in the range of 0 to 40.0 ng/mL were 0.999.And recoveries were stevioside 90.5%-94.9%,steviolbioside 87.7%-94.6%,glycyrrhizic acid 86.9%-97.3%and glycyrrhetinic acid 87.1%-96.5%at 50.0,200 and 400μg/kg,respectively.The precisions of stevioside,steviolbioside,glycyrrhizic acid and glycyrrhetinic acid were 2.21%,2.00%,1.88%and 2.72%,respectively.The method had the advantages of convenient operation,strong qualitative ability,high sensitivity and high accuracy.It was suitable for the detection of stevioside,steviolbioside,glycyrrhizic acid and glycyrrhetinic acid in soy sauce.