蛋黄中胆固醇含量测定方法的研究

Study on Determination Method of Egg Yolk Cholesterol Content

比较研究了高效液相色谱法(HPLC法)、皂化-比色法和试剂盒法3种测定蛋黄胆固醇含量的方法,结果表明,HPLC法能够准确、快速的分析出蛋黄中的胆固醇含量,较皂化-比色法和试剂盒法的准确度高。HPLC法测定蛋黄胆固醇含量的前处理为:用95%乙醇处理蛋黄稀释液,再用乙醚和石油醚混合萃取出胆固醇,用氮气吹干后加无水乙醇溶解,经0.45μm有机微孔滤膜过滤后制成样品。色谱条件为:C18柱,乙腈-异丙醇(41,V/V)作为流动相,流速为0.8mL·min-1,柱温35℃,检测波长210nm,进样量10μL,出峰时间14.578min。采用外标法定量。

The essay contrasted and studied on three methods of determinating the egg yolk cholesterol content. The three methods were High performance liquid chromatography（HPLC）, saponification-colorimetric method, and kit method. The results showed that HPLC method can accurately and fast analysis cholesterol content in egg yolk. The accuracy of HPLC was better than saponification-colorimetric method and kit method. The pre-treatment methods was using 95% ethanol dispose yolk diluent, then using ether and petroleum ether mixed extract cholester- ol, afterwards blow dry with nitrogen and add anhydrous alcohol to made dissolve, filtrate with 0.45 μm organic microporous membrane filter, so the sample is made. Chromatographic conditions was using C18 column, acetonitrile-isopropanol（4： 1,v/v） as mobile phase, flow velocity was 0.8 mL-min-1, column temperature was 35 ℃ detected wavelength was 210 nm, sample amount was 10 μL, the peak time was 14.578 min. Test used the external standard method quota.