正相色谱法测定大豆磷脂中的磷脂酰胆碱

Determination of phosphatidylcholine from soybean phospholipids by normal phase chromatography

目的建立正相色谱法测定大豆磷脂中磷脂酰胆碱含量,用于大豆磷脂类保健食品生产过程中该物质含量的测定和质量控制。方法采用硅胶柱(4.6 mm×250 mm,5.0μm)进行分离;色谱条件为流动相:异丙醇:正己烷:蒸馏水=70:16:14(V:V:V),等度洗脱,流速:1.0 m L/min;柱温:40℃;进样量:10μL;检测波长:205nm;并进行方法学验证,考察方法检测限、精密度、回收率等指标。结果磷脂酰胆碱峰面积与其浓度呈良好的线性关系,线性范围为0.1~0.8 mg/m L(r=0.9999),检出浓度0.04μg/m L,检出限为20μg/g,定量限为66.666μg/g,精密度(RSD)为0.6%,平均回收率99.37%(n=9)。结论本文建立的方法灵敏度高,重复性好,且具有很好的专属性,能够应用于大豆磷脂类保健食品中磷脂酰胆碱含量的检测。

Objective To establish a method for the determination and quality control of phosphatidylcholine from soybean phospholipids in health food by normal-phase chromatography. Methods Chromatographic conditions for determination phosphatidylcholine were silica gel column (4.6 mm×250 mm, 5 μm), isocratic elution with isopropanol:n-hexane:water=70:16:14 (V:V:V) as mobile phase, the flow rate of 1.0 mL/min, the column temperature of 40 ℃, the sample volume of 10 μL and the detection wavelength of 205 nm. The methodology validation was used to investigate method detection limit, precision, recovery, etc. Results The results showed that phosphatidylcholine had a good linear relationship at the range of 0.1~0.8 mg/mL between peak area and concentration (r=0.9999). The detection concentration was 0.04 μg/mL, the LOQ was 20 μg/g, the DOQ was 66.666 μg/g, the RSD was 0.6%and the average recovery rate was 99.37%(n=9). Conclusion The method is simple, accurate, sensitive and with good reproducibility by the method validation. It can be used for the determination of phosphatidylcholine from soybean phospholipids.