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目的:建立酰胺基亲水作用色谱-高效液相色谱-紫外法测定DHA颗粒中磷脂酰丝氨酸的含量。方法:以正己烷-异丙醇为提取溶剂对样品进行超声提取;选用Waters XBridge Amide(4.6 mm×250 mm,3.5μm)色谱柱,以乙腈-异丙醇(7∶3)为流动相,流速1.0 m L·min~(-1),柱温45℃,进样量20μL,紫外检测波长203 nm,单次运行时间10 min。结果:样品中磷脂酰丝氨酸获得很好的分离,在0.1~1.0mg·m L~(-1)线性关系良好(r=0.999 9);在高、低浓度的回收率(n=6)分别为101.2%和95.8%,RSD分别为1.3%和2.6%;3批样品中磷脂酰丝氨酸含量分别为10.5、10.3、10.6 mg·g~(-1)。结论:本方法操作简捷、环保、高效,经方法学验证,本法专属性、重复性好,准确度高,适应性强,为含磷脂酰丝氨酸功能食品的全面质量控制提供一种新的方法。
OBJECTIVE: To establish an amide-based hydrophilic interaction chromatography for the determination of phosphatidyl serine in DHA granules by high performance liquid chromatography-ultraviolet method. Methods: The sample was extracted by n-hexane-isopropanol. The sample was separated on a Waters XBridge Amide (4.6 mm × 250 mm, 3.5 μm) column with acetonitrile-isopropanol (7: The flow rate was 1.0 m L · min -1, the column temperature was 45 ℃, the injection volume was 20 μL, the UV detection wavelength was 203 nm, and the single run time was 10 min. Results: Phosphatidylserine was well separated in the samples with good linearity (r = 0.999 9) at 0.1-1.0 mg · mL -1 and recovery at high and low concentrations (n = 6) Were 101.2% and 95.8%, respectively. The RSDs were 1.3% and 2.6% respectively. The contents of phosphatidylserine in the three batches of samples were 10.5, 10.3, and 10.6 mg · g -1, respectively. Conclusion: The method is easy to operate, environment-friendly and efficient. The method is validated by methodology. The method has the characteristics of specificity, repeatability, accuracy and adaptability. It provides a new method for the total quality control of phosphatidylserine containing functional foods .