中国传统豆制品中异黄酮的超高效液相色谱紫外检测器快速定量法
Rapid quantification of isoflavones in traditional Chinese soybean food by ultra-high performance liquid chromatography coupling to the UV detector
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摘要: 建立了超高效液相色谱-紫外检测器(UPLC-UV)测定豆制品中大豆苷、黄豆黄苷、染料木苷、大豆苷元、黄豆黄素、染料木素的检测方法。采用酸化提取条件,将豆制品中大豆异黄酮及其修饰物水解成3种葡萄糖苷和3种苷元。采用BEH C18(2.1 mm×50 mm,1.7 μm)色谱柱,以0.1%甲酸和乙腈为流动相进行梯度洗脱,梯度洗脱程序为:0~10 min,10%~45%乙腈;10~12 min,45%~100%乙腈。流速为0.3 mL/min,柱温45℃,在波长260 nm处进行紫外检测。6种大豆异黄酮组分在7.5 min内达到完全分离。建立了外标校正标准曲线(R2 ≥ 0.9998),检出限在0.05~0.1 mg·L-1之间,加标回收率在96.8%~102.0%之间。利用本方法对腐竹、豆干、素百叶、素鸡、嫩豆腐、老豆腐和内酯豆腐等7种传统豆制品中大豆异黄酮进行了定性定量分析,总大豆异黄酮含量在8.67~25.83 g/kg间。不同豆制品间6种大豆异黄酮中以大豆苷和染料木苷为主要成分,占88.0%~93.4%。结果表明,本研究建立的UPLC法可有效降低杂质干扰,色谱基线稳定且峰型良好,在10 min内实现对6种大豆异黄酮的快速定量检测,可良好应用于常见市售豆制品的营养评估与质量控制。Abstract: An efficient and rapid quantitative method was developed for the evaluation of six types of isoflavones in soy food, including daidzin, glycitin, genistin, daidzein, glycitein and genistein. An UPLC column, BEH C18(2.1 mm×50 mm, 1.7 μm)was employed with acetonitrile and 0.1% acidic H2O(v/v)as mobile phase. The gradient was from 10% acetonitrile to 45% acetonitrile during 10 min at a flow rate of 0.3 mL/min. The detection wavelength was selected as 260 nm and the column was operated at 45℃. Six standard isoflavones were separated in 7.5 min and good linear relationships were achieved with correlation coefficients of 0.9998~0.9999. The recovery values in spiked samples were in the range of 96.8%~102.0%.The method was applied in the detection of isoflavones in traditional Chinese soyfood, including bean curd stick, dried toufu, bean curd shin, braised deep-fied bean curd, soft tofu, firm tofu and lactone bean curd. 8.67~25.83 g/kg isofavones was detected in the tested soyfood, and up to 88.0%~93.4% of isoflavones components were daidzin and genistin. These results suggested that isoflavones could be rapidly determined by ultra-high performance liquid chromatography coupled with photo-diode array detector in traditional Chinese soy food.