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目的:建立了测定氨基酸的在线衍生化方法。方法:以邻苯二甲醛(OPA)和9-芴甲基氯甲酸酯为衍生化试剂,对其氨基酸进行在线衍生化色谱分析。色谱柱:Zorbax Eclipse-AAA氨基酸分析柱(4.6 mm×150 mm,5μm),流动相A为40mmol·L-1Na2HPO4(用8 mol·L-1氢氧化钠溶液调pH 7.8),流动相B为乙腈-甲醇-水(45∶45∶10)梯度洗脱,流速2 mL·min-1,检测波长338,262 nm,柱温40℃。结果:氨基酸浓度在一定范围内,此法可以测定27种氨基酸的含量,但不适合清开灵注射液中氨基酸的含量测定。结论:该方法不能对清开灵注射液中氨基酸进行准确定量。
Objective: To establish an online derivatization method for determination of amino acids. Methods: The derivatization of phthalic acid (OPA) and 9-fluorenylmethyl chloroformate was used to derive their amino acids by on-line derivatization. Column: Zorbax Eclipse-AAA amino acid analytical column (4.6 mm × 150 mm, 5 μm), mobile phase A was 40 mmol·L-1Na2HPO4 (adjusted to pH 7.8 with 8 mol·L-1 sodium hydroxide solution) Acetonitrile-methanol-water (45:45:10). The flow rate was 2 mL · min-1. The detection wavelength was 338,262 nm and the column temperature was 40 ℃. Results: Amino acid concentration in a certain range, this method can determine the content of 27 kinds of amino acids, but not suitable for Qingkailing injection amino acid content determination. Conclusion: This method can not accurately quantify the amino acids in Qingkailing Injection.