Rapid separation and quantitation of combined neutral and polar lipid classes by high-performance liquid chromatography and evaporative light-scattering mass detection
Rapid separation and quantitation of combined neutral and polar lipid classes by high-performance liquid chromatography and evaporative light-scattering mass detection
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DOI：
10.1016/s0378-4347(97)00651-8
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发表时间：
1998-04-24
期刊：
影响因子：
--
通讯作者：
Anderson, MK
中科院分区：
文献类型：
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作者：
Homan, R;Anderson, MK
本文描述了一种创新的、广泛使用的高效液相色谱技术的改进，该技术可以通过蒸发光散射检测来解决非常广泛的脂质定量问题。在溶剂梯度程序的一部分中，丙酮取代2-丙醇产生一致的二酰基甘油和胆固醇的分辨率，而不牺牲其余主要脂类的基线分辨率。此外，消除了先前注意到的三酰甘油保留时间的不稳定性。丙酮的引入还可以在不增加总运行时间的情况下将流量降低20%。作为流动相组成的进一步修饰，乙酸和乙醇胺取代了最初显示的提高色谱柱性能的丝氨酸-乙胺组合。醋酸和乙醇胺的组合产生相同的结果，但这些溶质在丝氨酸上的挥发性增加导致基线噪声降低。最后，引入1,2-十六烷二醇作为内标，非常适合于该方法。通过对大鼠肝、心、肾和脑脂质提取物的成分分析，证明了这些修饰所获得的色谱性能。(C) 1998爱思唯尔科学有限公司
Modifications are described for an innovative and widely used high-performance liquid chromatography technique that resolves a very broad spectrum of lipids for quantitation by evaporative light-scattering detection. Substitution of acetone for 2-propanol in a portion of the solvent gradient program yields consistent resolution of diacylglycerol and cholesterol without sacrificing baseline resolution of the remaining major lipid classes. Moreover, previously noted instabilities in triacylglycerol retention time are eliminated. The introduction of acetone also enables a 20% reduction in flow-rate without an increase in total run time. As a further modification of the mobile phase composition, acetic acid and ethanolamine are substituted for the serine-ethylamine combination that was originally shown to improve column performance. The combination of acetic acid and ethanolamine yields the same result but the increased volatility of these solutes over serine results in decreased baseline noise. Finally, 1,2-hexadecanediol is introduced as an internal standard that is well suited for this method. The chromatographic performance obtained with these modifications is demonstrated in compositional analyses of lipid extracts from rat liver, heart, kidney and brain. (C) 1998 Elsevier Science B.V.