水解蛋白注射液中游离氨基酸含量的测定及分析
Determination and evaluation of amino acids in protein hydrolysate injection
分类号:
出版年·卷·期(页码):2012,32 (1):0-0
DOI:
10.16155/j.0254-1793.2017.01.01
-----摘要:-------------------------------------------------------------------------------------------
目的: 建立二硝基氟苯(DNFB)柱前衍生化法测定水解蛋白注射液中游离氨基酸含量,并对水解蛋白注射液质量进行分析。 方法: 采用DNFB为衍生化试剂进行衍生,色谱柱为Diamonsil C18(250 mm×4.6 mm,5 μm),流动相为0.05 mol·L-1醋酸钠溶液(含1% N,N-二甲基甲酰胺,调pH为6.4)与50%乙腈水溶液,梯度洗脱,流速1.0 mL·min-1,检测波长360 nm。 结果: 各氨基酸的线性关系良好;平均回收率(n=6)为98.7%~100.8%,RSD为0.46%~3.3%。 结论: 本研究建立了水解蛋白注射液游离氨基酸测定方法,为质量标准中游离氨基酸含量测定限度的制定提供了依据。
-----英文摘要:---------------------------------------------------------------------------------------
Objective: To establish a pro-column derivatization procedure for determination of the content of amino acid in protein hydrolysate injection by RP-HPLC. Methods: DNFB was used as a derivatization reagent and separation of DNP-amino acid derivatives were performed on Diamonsil C18 column(250 mm×4.6 mm,5 μm). The mobile phase was 0.05 mol·L-1 sodium acetate solution(containing 1% DMF and adjusted pH 6.4) and 50% acetonitrile with gradient elution at a flow rate of 1.0 mL·min-1, and the signals were detected at 360 nm. Results: Under the above condition, all amino acid in the standard solution were separated to baseline; The calibration curves of amino acids were in good linearity; The average recovery of amino acid ranged from 98.7% to 100.8% with RSD of 0.46%-3.3%(n=6). Conclusion: The method is applied to analyzing amino acid composition of protein hydrolysate injection and good results are obtained.
-----参考文献:---------------------------------------------------------------------------------------
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