摘要
目的建立测定虫草头孢菌粉中16种氨基酸含量的反相高效液相色谱(RP-HPLC)法。方法样品以6 mol.L-1盐酸于110℃水解24 h,内标为正亮氨酸,以异硫氰酸苯酯(PITC)为衍生化试剂,衍生后进行HPLC分析。采用Agela Venusil XBP C18(150 mm×4.6 mm,5μm)色谱柱,柱温为40℃;流动相A为0.1 mol.L-1醋酸-醋酸钠缓冲液(pH 6.5)-乙腈(95∶5),流动相B为乙腈-水(4∶1),梯度洗脱,流速为1.0 mL.min-1;检测波长为254 nm。结果 16种氨基酸浓度在1.81～7.50μg.mL-1范围内呈良好的线性关系(r=0.998 3～1.000 0);平均回收率除组氨酸、甲硫氨酸分别为92.9%和90.6%外,其余14种氨基酸在95.3%～104.1%之间;RSD除组氨酸、甲硫氨酸分别为2.8%和3.3%外,其余均<2.0%。结论该方法灵敏、准确,具有良好的重复性和稳定性,可用于虫草头孢菌粉中氨基酸的检测。
Objective To establish a quantitative method for detecting 16 amino acids in cordyceps mycelium powder. Methods The sample was hydrolyzed for 24 b at 110℃, and derived with phenyl isothiocyanate （ PITC ）, with norleueine serving as an internal standard. High performance liquid chromatography （HPLC） was performed on a Agela Venusil XBP Cls （ 150 minx4.6 ram,5 μm） column with gradient elution of 0.1 mol ·L-1 acetic acid-sodium acetate buffer solution （ pH 6.5 ） - aeetonitrile （95 ： 5 ） （A） and acetonitrile-water （4 ： 1 ） （B） at a flow rate of 1.0 mL · min-1, detected at 254 nm. Results Sixteen amino acids had a good linearity in the range of 1.81-7.50 μg ·mL-1 （ r = 0. 998 3-1. 000 0）. The average recoveries were 95.3% 104.1% , except His 92.9 % , Met 90.6% , and RSD less than 2.0% , except His 2.8% and Met 3.3%. Conclusion The method is sensitive and accurate with high repeatability and stability, and is helpful for the determination of amino acids in cordyceps mycehum powder.
出处
《医药导报》
CAS
北大核心
2012年第12期1624-1627,共4页
Herald of Medicine
