论文部分内容阅读
目的比较3种常用的组织三酰甘油(TG)、总胆固醇(TC)抽提方法。方法脂质提取分别采用3种方法。A:氯仿-甲醇抽提,B:庚烷-异丙醇-聚山梨酯20抽提,C:叔醇-Triton X-100-甲基乙醇抽提。采用氧化酶法检测混合人血清、普食或高脂饲养大鼠肝组织TG和TC水平,自动生化仪或手工比色测定。结果在3种方法中,A法较其他方法耗时长;C法需手工测定。尽管3种方法的批内和批间差异无明显不同,方法A、B提取的TG、TC回收率随抽提溶液体积增大而增加,而C法回收率随抽提液体积增大而呈下降趋势。采用B法作组织抽提,普食大鼠肝脏中TG、TC的抽提量不随抽提液体积的改变而变化,高脂大鼠肝脏TG、TC含量随抽提液体积增加而升高。进一步选用5 mL抽提液提取普食和高脂大鼠肝脏,高脂大鼠TG、TC含量均明显高于普食组(P<0.01)。结论 3种脂质抽提方法中,B法回收率高,自动化程度高、省时,适合肥胖大鼠肝脏脂质的抽提。用B法提取50 mg高脂肝脏脂质时,抽提溶液体积以5或7 mL为宜。
Objective To compare the three commonly used tissue triglyceride (TG) and total cholesterol (TC) extraction methods. Methods Lipid extraction were used three methods. A: chloroform-methanol extraction, B: heptane-isopropanol-polysorbate 20 extraction, C: tertiary alcohol-Triton X-100-methyl ethanol extraction. Oxidase method was used to detect the levels of TG and TC in mixed human serum, normal diet or high fat diet rat liver tissue, automatic biochemical analyzer or manual colorimetric assay. Results Of the three methods, method A took longer than other methods; method C was determined manually. Although there was no significant difference between in-batch and inter-batch differences among the three methods, the recoveries of TG and TC extracted by methods A and B increased with the increase of the volume of extraction solution, while the recovery of C was increased with the volume of extraction solution Downtrend. Using the B method for tissue extraction, the extraction amount of TG and TC in the liver of the normal diet did not change with the volume of the extraction liquid. The contents of TG and TC in the liver of the high-fat diet increased with the volume of the extraction liquid. The contents of TG and TC in hepatic and hyperlipidemic rats fed with 5 mL extract were significantly higher than those in normal diet group (P <0.01). Conclusion Among the three lipid extraction methods, the recovery rate of B method is high, the degree of automation is high, and time-saving is suitable for the extraction of liver lipids in obese rats. When using B method to extract 50 mg of high-fat liver lipids, it is advisable to extract the solution in a volume of 5 or 7 mL.