Analysis and Evaluation of 4 Main Amino Acids without Derivatization in Colla Corii Asini and Donkey-Hide Gelatin Juice by RP-HPLC
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摘要: 建立了一种高效液相色谱-蒸发光散射检测法(HPLC-ELSD)同时测定阿胶和阿胶汁中4种主要未衍生化氨基酸(L-甘氨酸、L-羟脯氨酸、L-丙氨酸、L-脯氨酸)含量的方法,从源头对阿胶进行质量控制。采用6 mol·L-1的盐酸对经真空干燥后的阿胶及阿胶汁粉末110℃恒温水解24 h,以Thermo hypercarb(150 mm×4.6 mm,5 μm)为分析柱,流动相A为0.08%三氟乙酸甲醇:乙腈(90:10),B为2 mmol·L-1全氟戊酸水,梯度洗脱,流速为0.8 mL·min-1。结果表明,L-甘氨酸、L-羟脯氨酸、L-丙氨酸、L-脯氨酸的质量浓度与峰面积呈良好的线性关系(r>0.9993),阿胶与阿胶汁的加标回收率分别为97.42%~103.29%、97.17%~102.19%,RSD均<2.46%(n=6)。此方法简单快速、重复性较好,适用于快速准确地测定阿胶和阿胶汁中L-甘氨酸、L-羟脯氨酸、L-丙氨酸、L-脯氨酸含量,为其质量控制提供参考。Abstract: An analytical method for the determination of non-derivatized amino acids by HPLC-ELSD was developed. The contents of L-glycine(Gly),L-hydroxyproline(Hyp),L-alanine(Ala)and L-proline(Pro)in Colla Corii Asini and donkey-hide gelatin juice were determined simultaneously. The dried Colla Corii Asini and donkey-hide gelatin juice powder were hydrolyzed with 6 mol·L-1 hydrochloric acid at 110℃ for 24 h and then analyzed by Thermo hypercarb(150 mm×4.6 mm,5 μm)with methanol-acetonitrile solution containing 0.08% trifluoroacetic acid(90:10)as mobile phase A and water containing 2 mmol·L-1 perfluorovalerate as mobile phase B with a flow rate of 0.8 mL·min-1 under the gradient elution. The carrier gas flow rate was 1.5 L·min-1 and the temperature of drift tube was 55℃. As the results,the four amino acids were completely separated and there was a good linearity in the certain range(r>0.9993). The average recoveries of four amino acids were 97.42%~103.29% and 97.17%~102.19% in the donkey-hide gelatin and donkey-hide gelatin juice respectively with RSD<2.46%(n=6). The developed method was simple,precise and good reproducibility. It could be used to quickly and accurately determine the content of four underivatized amino acids in donkey-hide gelatin and donkey-hide gelatin juice,which was suitable for the quality control of Colla Corii Asini.
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Keywords:
- Colla Corii Asini /
- donkey-hide gelatin juice /
- amino acid /
- non-derivatized /
- HPLC-ELSD
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期刊类型引用(3)
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2. 吴崇乐,颜冬兰,黄胜,罗菲菲,叶惠煊. 阿胶及其制剂质量控制方法研究进展. 海峡药学. 2022(04): 1-4 . 百度学术
3. 邹惠亮,陈褚建,王新财,潘渭樵,陈柯,沈丽,陈敏. UPLC-MS/MS法快速测定鳖甲胶中4种主要氨基酸的含量. 中国药品标准. 2022(05): 535-540 . 百度学术
其他类型引用(2)
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