超临界流体色谱法测定蛋黄卵磷脂中的8种磷脂组分

Determination of eight phospholipid components in egg yolk lecithin by supercritical fluid chromatography

目的:建立超临界流体色谱法分离并测定蛋黄卵磷脂中的三棕榈酸甘油酯(TG)、胆固醇(CH)、磷脂酰乙醇胺(PE)、溶血磷脂酰乙醇胺(LPE)、磷脂酰肌醇(PI)、磷脂酰胆碱(PC)、鞘磷脂(SM)、溶血磷脂酰胆碱(LPC)8种磷脂组分。方法:采用Zorbax Silica色谱柱(250 mm×4.6 mm,5μm)与Li Chrosphere 100 Diol色谱柱(250 mm×4 mm,5μm)串联,以二氧化碳为流动相A,甲醇-氨水(100∶0.5)为流动相B,进行梯度洗脱,流速为2.3 m L·min^(-1),柱温为30℃,进样量为5μL,二氧化碳补偿液(甲醇)流速为0.2 m L·min^(-1),蒸发光散射检测器检测,漂移管温度为90℃,雾化气流速为1.2 L·min^(-1)。结果:8种磷脂组分分离完全,且峰面积与浓度的双对数线性关系良好,日内精密度RSD为0.2%~2.0%,日间精密度RSD为0.5%~2.0%,检测限为0.137~0.584μg,定量限为0.273~1.169μg,加样回收率为97.1%~100.6%,RSD为1.1%~2.7%,测得3批蛋黄卵磷脂中的TG为1.6%~1.8%,CH为0.4%~0.5%,PE为7.8%~8.9%,LPE为0.7%~0.9%,PI为0.9%~1.1%,PC为79.1%~83.0%,SM为1.4%~1.5%,LPC为1.6%~1.7%。结论:本文建立的方法可用于蛋黄卵磷脂的质量控制。

Objective： To establish a supercritical fluid chromatography method for separation and determination of glycerol tripalmitatec （ TG ）, cholesterol （ CH ）, photidylethanolamine （ PE ）, lysophosphatidyl ethanolamine （ LPE ）, phophotidyl inositol （ PI ）, phosphotidylcholine （ PC ）, sphingomyelin （ SM ） and lysophosphatidyl choline （ LPC ） in egg yolk lecithin. Methods： A Zorbax Silica column （ 250 mm × 4.6 mm, 5μm ） and a LiChrosphere 100 Diol column （ 250 mm × 4 mm, 5 μm ） in series were used with the mobile phase A of carbon dioxide and the mobile phase B of methanol-ammonia solution （ 100 ： 0.5 ） by gradient elution. The flow rate was 2.3 mL· min^-1 , and the column temperature was controlled at 30℃. The injection volume was 5 μL, and the flow rate of carbon dioxide makeup （ methanol ） was 0.2 mL· min^-1. The samples were detected by evaporative light scattering detector with drift tube temperature at 90 ℃ and a flow of atomizing gas at 1.2 L· min^-1. Results： Eight phospholipid components were separated completely. The double logarithm linear relationship of peak area and corresponding concentration was good. Within-day and intra-day precision of the method were 0.2%-2.0% and 0.5%-2.0%, respectively. The limit of detection was 0.137-0.584μg, and limit of quantification was 0.273- 1.169 μg. The spiked sample average recoveries were 97.1%-100.6%, and the RSD was 1.1%-2.7%. Eight phospholipid components were determined in 3 batch of egg yolk lecithin. TG were 1.6%-1.8%, CH were 0.4%- 0.5%, PE was 7.8%-8.9%, LPE was 0.7%-0.9%, PI was 0.9%-1.1%, PC were 79.1%-83.0%, SM was 1.4%- 1.5%, and LPC were 1.6%-1.7%, respectively. Conclusions： The proposed method is proved method validation, and thus can be used to control the quality of egg yolk lecithin.