酶标仪分光光度法定量测定鹰嘴豆总皂苷

Determination of Chickpea Saponins Content by Colorimetry Spectrophotometry

借鉴比色法定量测定大豆皂苷的方法,采用香草醛-高氯酸-冰乙酸体系显色,再采用酶标仪结合三波长法测定和校正吸光值,通过对提取和测定方法的优化,拟建立一套适用于测定鹰嘴豆总皂苷的三波长分光光度法。结果表明,鹰嘴豆总皂苷最佳提取条件是:脱脂后取1.00 g鹰嘴豆粕加入25 mL 70%的乙醇,70℃水浴提取3 h;皂苷一次提取效率为95.09%,两次合并提取效率为98.51%;精密度试验的变异系数CV为3.37%,平均加标样回收率为95.57%,变异系数CV为3.221%;皂苷显色后的最大吸收峰会受到蛋白杂质的干扰,需采用HCl调节pH沉淀蛋白。所建立的测定方法具有操作简便、重现性好、精密度高,适于大批量样品测定以及产品生产的质量控制及监测等特点。

Base on the method of determinating soybean saponins by colorimetry, The HCLO4 - vanilin staining method was adopted and the saponin absorbency was determined by high performanc microplate spectrophotometer and revised by the three - wavelength method in the present study. The extraction and determination were optimized to build a determining method of the total saponins content of chickpea seeds by three - wavelength colorimetry spectrophotometry. The optimum extraction conditions are 1 g chickpea meal, extracted with 70 % alcohol solution of 25 mL at temperatures 70 ℃ for 3 h. The saponin extraction rate is 95.09% for one time extraction and 98.51% total rate for twice extraction. In accuracy experiment the coefficient of variation （cv） is 3.37%, and the average recovery rate is 95.57% with cv being 3. 221%. Due to the interference of protein impurity to the saponin determination, HC1 solution is needed to adjust pH to deposit the protein. The newdeveloped determining method is reliable, simple and accurate, and is suitable for multi - sample determination and product quality control in production.