摘要
目的建立柱前手性衍生化-反相高效液相色谱法(HPLC)测定L-肌肽对映体的纯度。方法以邻苯二甲醛/N-乙酰基-L-半胱氨酸(OPA/NAC)为手性衍生化试剂,反应生成具有荧光吸收的一对非对映异构体衍生物,采用C18色谱柱(250mm×4.6mm,5μm),流速1.0mL·min-1,50mmol·L-1乙酸铵缓冲溶液(pH6.0)-甲醇(70:30,V/V)流动相,柱温35℃,λex=350nm,λem=450nm。结果L-肌肽与其光学异构体分离度大于3,在0.104～2.68mg·L-1范围内,D-肌肽衍生物色谱峰面积与其浓度呈良好线性关系,检测限浓度为0.02mg·L-1。结论建立的L-肌肽光学异构体(杂质)手性衍生化-HPLC拆分检查法方便准确,可用于L-肌肽的光学纯度控制。
AIM To establish a chiral precolumn derivatization method for testing the optical purity of Lcarnosine.METHODS Enantiomers as a pair of non-chiral derivative was derivatized from ortho-phthalaldehyde /N-acety-L-cysteine （OPA /NAC） as chiral derivatization agent which was carried out on a C18 （250 mm × 4.6 mm,5 μm） column at 35 ℃ with a mixture of 50 mmol·L-1 acetate buffer （pH=6.0）-methanol （70：30,V /V） as mobile phase with a flow rate of 1.0 mL·min-1;with fluorescence detector at λex =350 nm and λem=450 nm.RESULTS The separation grade was greater than 3.0 between L-carnosine and its enantiomers within the range of 0.104 to 2.68 mg·L-1.The peak chromato-area of D-carnosine and its concentration showed good linear correlation,and the limit of detection （LOD） was 0.02 mg·L-1.CONCLUSION The established chiral derivatization HPLC testing method is simple and accurate for the optical purity control of L-carnosine.
出处
《中国新药与临床杂志》
CAS
CSCD
北大核心
2010年第1期58-61,共4页
Chinese Journal of New Drugs and Clinical Remedies
关键词
肌肽
色谱法
高压液相
抗氧化剂
光学纯度
对映体
手性拆分
柱前衍生化
carnosine
chromatography
high pressure liquid
antioxidants
optical purity
enantiomers
chiral separation
pre-column derivation
