超高效液相色谱法测定原料药中加米霉素的含量

Determination of Gamithromycin in Drug Substance by UPLC

关键词:  加米霉素  超高效液相色谱法  二极管阵列检测器

Key words:  gamithromycin  ultra-high performance liquid chromatography  photodiode array detector

摘要:

研究建立超高效液相色谱-光电二极管阵列法（UPLC-PDA）测定加米霉素原料中主成分加米霉素的含量。采用反相超高效液相色谱法对加米霉素进行色谱分离和快速定量测定。方法采用ACQUITY UPLCTM CSH C18色谱柱（2.1 mm×100 mm,1.7 μm)为分离柱，柱温：35℃；以氨水(0.1:30)-乙腈为流动相（30:70）进行等度洗脱；流速：0.45 mL/min；进样量：1 μL；检测波长为208 nm。通过光谱图、保留时间和峰面积参数对加米霉素进行定性、定量检测。加米霉素在80~1600 μg/mL的范围内线性关系良好（R2=0.9996）；方法检测限为40 μg/mL、定量限为80 μg/mL，系统适应性试验色谱柱耐受性良好，添加回收率99.80%，完全满足检测需求。研究建立的UPLC方法色谱分离较好，分析速度较快，前处理简单，适用于加米霉素原料中主成分加米霉素含量的检测。

Abstract:

In this study, a ultra-high performance liquid chromatography- photodiode array method (UPLC-PDA) was established to determine the content of gamithroycin. Reversed-phase HPLC method was used for chromatographic separation and rapid quantitative determination of gamithromycin. ACQUITY UPLCTM CSH C18 column (2.1 mm×100 mm, 1.7 μm) was used as separation columns, Column temperature :35 ℃; Ammonia water (0.1:30): acetonitrile was used as mobile phase (30:70) for equal elution; Flow rate was 0.45 mL/min; Volume of injection was 1μL; Detection wavelength was 208 nm. Gamithromycin was qualitatively and quantitatively detected by spectrogram, retention time and peak area parameters. The linear relationship of gamithromycin was good in the range of 80~1600μg/mL (R2=0.9997); The limit of detection of the method is 40μg/mL,the quantitation of the method is 80μg/mL, the chromatographic column of system adaptability test was well tolerated, and the recovery rate was 99.80%,which fully meet the testing requirements. This method has better chromatographic separation, faster analysis, simple pre-processing, It is suitable for the detection of the content of the main component of gamithromycin.