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目的建立一种采用反相高效液相色谱法测定羊胎盘枸杞胶囊中16种氨基酸的含量。方法样品中的蛋白质在6 mol/L盐酸溶液中水解为氨基酸,其中一级氨基酸用邻苯二甲醛(OPA)、二级氨基酸用9-芴甲基氯甲酸酯(FMOC)进行柱前在线衍生;用Agilent Hypersil AA-ODS柱分离;用流动相(0.16%醋酸钠)进行梯度洗脱;经荧光检测器(FLD)检测(激发波长:340 nm,发射波长:450 nm;15 min时,激发波长:266 nm,发射波长:305 nm)。结果 16种氨基酸在5~250 pmol/μL浓度范围内呈现良好的线性关系(r:0.99959~1.00000,n=5),平均加标回收率为91.6%~104.1%(n=9),相对标准偏差(RSD)为0.9%~3.7%,最低检测限为0.4~1.2 pmol。结论该方法灵敏度高,具有良好的精密度与准确度,可以有效地控制产品质量。
Objective To establish a method for the determination of 16 kinds of amino acids in sheep placenta goji capsules by reversed-phase high performance liquid chromatography. Methods The proteins in the sample were hydrolyzed to amino acids in 6 mol / L hydrochloric acid solution. The first amino acid was hydrolyzed with OPA and the second amino acid with 9-fluorenyl methyl chloroformate (FMOC) (Gradient elution: 340 nm, emission wavelength: 450 nm). At 15 min, the sample was separated on an Agilent Hypersil AA-ODS column and eluted with a mobile phase (0.16% sodium acetate) Excitation wavelength: 266 nm, Emission wavelength: 305 nm). Results The 16 amino acids showed good linearity (r: 0.99959 ~ 1.00000, n = 5) in the range of 5 ~ 250 pmol / μL with the average recoveries of 91.6% -104.1% (n = 9) Deviation (RSD) was 0.9% ~ 3.7%, the lowest detection limit was 0.4 ~ 1.2 pmol. Conclusion The method has high sensitivity, good precision and accuracy, and can effectively control product quality.