高效液相色谱法测定人尿中神经鞘氨醇和二氢神经鞘氨醇  

Determination of Sphingosine and Sphinganine in Human Urine by High Performance Liquid Chromatography

作　　者：邱茂锋[1] 刘秀梅[1] 王玉华[1] 刘江[1] 

机构地区：[1]中国预防医学科学院营养与食品卫生研究所,北京1000502

出　　处：《分析化学》2000年第4期473-475,共3页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金!39500121

摘　　要：建立了一种检测人尿中神经鞘氨醇（Ｓｏ）和二氢神经鞘氨醇（Ｓａ）的高效液相色谱方法（ＨＰＬＣ）。离心分离尿样中的片状剥落细胞，解后用乙酯萃取、邻苯二甲衍生，在训梯度洗脱用Ｎｏｖａ－Ｐａｋ Ｃ１８－ＲＰ色谱柱（１５ｃｍ×３．９ｍｍ，４μｍ）分离、荧光检测器检测。Ｓｏ、Ｓａ的检出限均为０．０５ｎｇ（女性尿样０．０７５μｇ／Ｌ，男性尿样０．００５μｇ／Ｌ）。分析从我国一个村庄采集的４０份尿样。An improved liquid chromatographic method for the determination of sphingosine(So) andsphinganin(Sa) in human urine was developed. It involves isolation of exfoliated cells from human urinefollowed by a rapid and efficient extraction of sphingolipid bases in ethylacetate, and a derivatization stepwith o-phthaldialdehyde and a high performance liquid chromatographic separation on a 15 cm × 3. 9mm, 4 μm Nova-Pak C18-RP column, with a phosphate buffer/methanol gradient. Fluorescence wasmonitored at 335 nm for excitation and 440 nm for emission. The limits of detection were 0.05 ng or0.075 μg/L in femal urine and 0.005 μg/L in male urine. The method was applied to analyze 40 humanurin samples collected from a village in China. The So, Sa and Sa/So ratio in female urine varied from1 .29 ~ 13.58μg/L, 0.25 ~ 3.13 μg/L and 0. 15 ~ 0.25, respectively, while those in male urine variedfrom 0.075 ~ 3.07 μg/L, 0.019 ~ 0.50 μg/L and 0.028 ~ 0.26, respectively.

关 键 词：神经鞘氨醇 二氢神经鞘氨醇 人尿 HPLC 伏马菌素 

分 类 号：O657.7[理学—分析化学]