中文摘要：

大豆异黄酮育种与大豆制品加工研究需要进行大批量样品12种异黄酮组分的快速定量分析。在前人研究基础上利用Agilent 1100高效液相色谱（HPLC）系统,以大豆品种南农95C-13为材料,研究大豆苷元、大豆苷、乙酰基大豆苷、丙二酰基大豆苷、染料木苷元、染料木苷、乙酰基染料木苷、丙二酰基染料木苷、黄豆苷元、黄豆苷、乙酰基黄豆苷、丙二酰基黄豆苷等12种标准品外标法快速定量技术。从样品制备与色谱条件对分析12种异黄酮组分的准确度和分离度入手,确定分析流程,以（科丰1号×南农1138-2）的184个重组自交系（NJRIKY）为材料,研究豆腐加工中总量和各组分的变化特点。（1）样品以80%甲醇水溶液50℃超声1 h提取;色谱条件为,检测波长254 nm,柱温36℃,流速2.0 mL min^-1,进样量10μL,流动相0.1%（V/V）乙酸水溶液（A）和100%甲醇（B）,02 min,27%B（V/V）→2-3 min,27%-38%B→310 min,38%B→1012 min,38%-39%B→1214 min,39%B→1415 min,39-27%B梯度洗脱;在15 min内将12个组分良好分离,各组分峰面积与其相应浓度均呈良好线性关系（R^2为0.99760.9999）;加标回收率均大于99%,变异系数低于2%。（2）NJRIKY群体籽粒、豆乳、豆腐异黄酮总量和组分的大量分析验证了HPLC快速技术的效果。籽粒异黄酮总含量（3 695.00μg g^-1）在豆乳加工中平均85.15%转入豆乳（3 146.12μg g^-1）,14.85%（548.88μg g^-1）进入豆渣。传统豆腐加工通过硫酸钙絮凝,只有17.32%（639.89μg g^-1）转入豆腐,67.83%（2 506.23μg g^-1）留在黄浆水中。豆乳中12种组分含量比籽粒中稍低,均以丙酰基染料木苷含量最高;而豆腐中乙酰基染料木苷和乙酰基黄豆苷缺失,以染料木苷元与大豆苷元含量较高。大豆科丰1号与南农1138-2杂交重组后家系间的异黄酮遗传变异增大,增加了对其遗传改良的潜力。

英文摘要：

A rapid,precise,and stable quantifying method of isoflavone components is the key to quality soy-food processing and genetic improvement of quality soybeans.A quick procedure used Agilent 1100 high performance liquid chromatograph（HPLC） system with the diode array detection（DAD） and Zorbax SB-C18 packed column（5 μm,4.6 ID × 150 mm） using external standards of isoflavone components,including daidzein,daidzin,6＂-O-acetyldaidzin,6＂-O-malonyldaidzin,genistein,genistin, 6＂-O-acetylgenistin,6＂-O-malonylgenistin,glycitein,glycitin,6＂-O-acetylglycitin and 6＂-O-malonylglycitin was established for measuring the 12 isoflavones in soybean seeds and its processing products.The procedure includes the following key points：80% methanol aqueous solution under ultrasonication for 1 h at 50℃ was chosen;for separation of the 12 isoflavone components within 15 min,the mobile phase of 0.1% acetic acid（V/V） aqueous solvent A and 100% methanol solvent B with the flow rate of 2.0 mL min^-1,injection volume at 10 μL,column temperature at 36℃ and detection wavelength at 254 nm were selected;and the linear gradient extraction of 0-2 min,27% B（V/V）→2-3 min,27-38% B→3-10 min,38% B→10-12 min,38-39% B→12-14 min,39% B→14-15 min,39%-27% B was adopted.The procedure was linear（R^2 = 0.9976-0.9999）,precise（CV or RSD ranged from 0.90% to 3.35% for 2 232 samples from NJRIKY）,accurate [recoveries were more than 99.00% for the different concentrations of the 12 isoflavones（CV of 0.22%-1.40%）],robust（inter-day CV of 0.24%-3.95%） and rapid（less than 15 min for 12 isoflavones resolved）.The procedure was verified to be effective by a large sample determination of isoflavone components in soybean seed,soymilk and tofu using the soybean population of NJRIKY.The data indicated that the isoflavones were 3 695.00 μg g^-1 in seed,among them 14.85%（548.88 μg g^-1） were transferred to the residual,85.15%（3 146.12 μg g^-1） to soymilk,but only 17.32%（639.89 μg g^-1） to tofu while 6