Zorn U, Wolf C F, Wennauer R, Bachem M G, Grünert A
Institute of Clinical Chemistry, University of Ulm, Germany.
Electrophoresis. 1999 Jun;20(7):1619-26. doi: 10.1002/(SICI)1522-2683(19990601)20:7<1619::AID-ELPS1619>3.0.CO;2-U.
Combining specific enzymatic derivatization of cholesterol or triglycerides with capillary isotachophoresis (CITP), human serum lipoproteins are separated into 14 lipoprotein subfractions, monitored and quantitated by direct capillary UV detection. By comparing the separation patterns of human serum with the patterns of lipoprotein particles isolated by sequential ultracentrifugation it became evident that peaks 1-5 represent lipoproteins of the high density lipoprotein (HDL) fraction, peaks 6-8 embody the very low density lipoprotein (VLDL) fraction and chylomicrons, and peaks 7-14 represent the low density lipoprotein (LDL) fraction. Peaks 7 and 8 were found in the VLDL as well as in the LDL fraction. Using triglyceride-specific staining peaks 6-8 occurred prominently; and with cholesterol-specific staining, peaks 1-5 and 7-14 were prominent. The coefficient of variation, for the sum of the peak heights of a pooled serum, was 3.94 for triglyceride-specific staining and 2.32 for cholesterol-specific staining. A linearity range between 0.23 and 2.29 mM/L was found for triglyceride-specific staining and between 0.043 and 4.33 mM/L for cholesterol-specific staining. The practicability of the method was evaluated (i) using blood of humans before and 45 min after an oral fat load. Triglyceride-specific staining revealed a prominent increase in the VLDL fraction and chylomicrones containing peaks 6 and 7, and a minor increase in the HDL fraction containing peaks 3 and 4, and (ii) in patients with manifest hypothyroidism before and after thyroxine therapy. Cholesterol-specific staining demonstrated a massive decrease in the first peak of the HDL fraction and in peaks 9 and 11 of the LDL fraction regarding the hypo versus hyperthyroid state.
将胆固醇或甘油三酯的特定酶促衍生化与毛细管等速电泳（CITP）相结合，可将人血清脂蛋白分离为14个脂蛋白亚组分，并通过直接毛细管紫外检测进行监测和定量。通过比较人血清的分离模式与通过连续超速离心分离的脂蛋白颗粒的模式，发现峰1 - 5代表高密度脂蛋白（HDL）组分的脂蛋白，峰6 - 8体现极低密度脂蛋白（VLDL）组分和乳糜微粒，峰7 - 14代表低密度脂蛋白（LDL）组分。在VLDL以及LDL组分中均发现了峰7和峰8。使用甘油三酯特异性染色时，峰6 - 8显著出现；而使用胆固醇特异性染色时，峰1 - 5和峰7 - 14显著。对于混合血清峰高总和，甘油三酯特异性染色的变异系数为3.94，胆固醇特异性染色的变异系数为2.32。甘油三酯特异性染色的线性范围为0.23至2.29 mM/L，胆固醇特异性染色的线性范围为0.043至4.33 mM/L。对该方法的实用性进行了评估：（i）使用人类口服脂肪负荷前和45分钟后的血液。甘油三酯特异性染色显示，包含峰6和峰7的VLDL组分和乳糜微粒显著增加，包含峰3和峰4的HDL组分略有增加；（ii）对明显甲状腺功能减退患者在甲状腺素治疗前后进行评估。胆固醇特异性染色表明，与甲状腺功能减退状态相比，HDL组分的第一个峰以及LDL组分的峰9和峰11在甲状腺功能亢进状态下大幅下降。
