摘要
目的建立测定肝组织核黄素含量的高效液相色谱方法。方法取肝组织样品加入0．1mol／L盐酸，用匀浆器研磨制备匀浆后，经酸解、10％胰蛋白酶水解处理，用微孔滤膜过滤后取滤液分析。采用AtlantisC18色谱柱（150mm×4．6mm，5μm）分离，以甲醇：5mmol／L醋酸铵（35：65，v／v）为流动相，流速为1．0ml／min，荧光检测器检测时激发波长为450nm，发射波长为520nm。结果本法测定核黄素的线性范围为1．0—200ng／ml，最低检测限为1．0ng／ml；日内和日间测定的核黄素标准品的峰面积的相对标准偏差分别为2．83％和3．76％，样品分别为0．71％和2．17％；核黄素在样品中的加标回收率为98．6％－101．1％。结论建立的高效液相色谱方法简便、准确、灵敏，可用于测定肝组织中核黄素的含量。
Objective To develop a high performance liquid chromatographic （HPLC） method for the determination of riboflavin in hepatic tissue. Methods The hepatic samples were grinded uniformly with homogenizer. The homogenate was then treated with 0. 1 mol/L HCL and 10% trypsin hydrolysis, and analyzed after filtration. An Atlantis C18 column （ 150 mm ×4. 6 mm, 5 μm） was used. The mobile phase consisted of 35% methanol and 65% 5 mmol/L ammonium acetate solution. The flow rate was 1.0 ml/min. The spectrophotofluorometer was set at a wavelength of 450 nm for excitation and 520 nm for emission. Results A good linear correlation between riboflavin concentration （ from 1.0 ng/mL to 200 ng/mL） and fluorescence intensity was established. The detection limit was 1.0 ng/mL. The standard substance of riboflavin and the samples in single-day and multi-day relative standard deviations were in the range of 2. 83%-3.76% and 0. 71%-2. 17%, respectively. The recoveries for riboflavin in hepatic tissue were in the ranges of 98. 6%-101.1%. Conclusion The established HPLC method is quick, accurate, and sensitive, and can be applied to the determination of riboflavin in hepatic tissue.
出处
《中华临床营养杂志》
CAS
2009年第4期231-234,共4页
Chinese Journal of Clinical Nutrition
基金
国家自然科学基金（30872097）
关键词
高效液相色谱法
肝组织
核黄素
High performance liquid chromatography
Hepatic tissue
Riboflavin
