[Determination of amino acids in Isatis indigotica Fort by reversed-phase high performance liquid chromatography with pre-column derivatization].
A reversed-phase high performance liquid chromatographic method with pre-column derivatization for the determination of amino acids, which were known as the characteristic constituents of Isatis indigotica Fort was established. The amino acids in alkalescence were derivatized with 2, 4-dinitro-fluorobenzene (DNFB). A reproducible method for simultaneous qualitative analysis of glutamic acid (Glu), argirine (Arg) and proline (Pro) and quantitative analysis of Arg and Pro in Isatis indigotica Fort has been established. NaAc buffer (pH 6.4) acetonitrile (85:15, v/v) as mobile phase and a Sinochrom ODS-BP column were used. The detector was operated at 360 nm. The linear regressions of the standard curves were determined for Arg and for Pro. The method was carried out over the range of 0. 627 - 5.016 microg for Arg and 0. 874 - 7.000 microg for Pro. The recoveries were 98.5% and 98.4% with the relative standard deviations of 2.5% and 2.3% for Arg and Pro respectively. The results indicate that among the three amino acids in Isatis indigotica Fort, Arg content was the highest, Pro the second and Glu the lowest. The method has good accuracy and repeatability and it can be used for the quality control of Isatis indigotica Fort.
Wan
Shaohui
S
Pharmaceutical College of Henan University, Kaifeng 475001, China. shaohuiw@126.com
Yang
Hao
H
Geng
Xiumei
X
D000596:Amino Acids / Q000032:analysis* / Q000737:chemistry*; D001120:Arginine / Q000032:analysis / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004139:Dinitrofluorobenzene / Q000737:chemistry; D018698:Glutamic Acid / Q000032:analysis / Q000737:chemistry; D031219:Isatis / Q000737:chemistry*; D011392:Proline / Q000032:analysis / Q000737:chemistry
D000596:Amino Acids; D018698:Glutamic Acid; D001120:Arginine; D011392:Proline; D004139:Dinitrofluorobenzene