HPLC测定保泰松原料及糖衣片剂有关物质的改进研究

Improvement Research of a Determination Method for the Related Substance in Crude and Preparation of Phenylbutazone by HPLC

关键词:  保泰松  有关物质  高效液相色谱法  杂质  保泰松片  杂质分离  质量控制  方法改进

Key words:  phenylbutazone  related substance  HPLC  impurity  phenylbutazone tablets  impurities separation  quality control  method improvement

摘要:

目的 建立保泰松原料及糖衣片剂中5个已知杂质及其他未知杂质检测方法。方法 分离条件：Agilent Eclipse XDB-C₁₈（4.6 mm×250 mm，5 μm），柱温为30℃，流动相为醋酸铵缓冲液（取醋酸铵2.72 g，加水700 ml溶解，用冰醋酸调节pH值至4.1，加水至1 000 ml，摇匀）-乙腈（58：42），流速为1.5 mL·min^-1，检测波长为254 nm，进样量为20 μl。结果 5个已知杂质在25 min内能够完全分离，其中杂质A、B、C、D均在5~30 μg·ml^-1，杂质E在0.03~0.15 μg·ml^-1具有良好的线性关系；最低检出限量分别为27.86，28.52，26.28，31.96，0.24 ng；原料、糖衣片剂中5个已知杂质的平均回收率分别为98.1%，99.3%，97.6%，97.4%，95.1%和96.9%，97.1%，96.6%，96.1%，94.7%。结论 改进的方法灵敏度更高、定量准确，重复性更好，可有效控制保泰松原料及糖衣片剂中杂质含量。

Abstract:

OBJECTIVE To establish a method to detect five kinds of known impurities and others unknown impurities in phenylbutazone crude drug and coated tablet. METHODS The HPLC analysis was performed on a Agilent Eclipse XDB-C₁₈(5 μm, 4.6 mm×250 mm)column; the column temperature was 30℃; the mobile phase consisted of ammonium acetate buffer (take ammonium acetate 2.72 g, dissolve 700 ml of water, adjust the pH to 4.1 with glacial acetic acid, add water to 1 000 ml, shake well) and acetonitrile(58:42) at the flow rate of 1.5 mL·min^-1, the detection wavelength was 254 nm, and injection volume was 20 μl. RESULTS Five kinds of impurities were completely separated in 25 min, and good linear relationships of A, B, C, D were obtained in the range of 5-30 μg·ml^-1. good linear relationships of E was obtained in the range of 0.03-0.15 μg·ml^-1, the limit of detection of were respectively 27.86, 28.52, 26.28, 31.96, 0.24 ng; the average recoveries were respectively 98.1%, 99.3%, 97.6%, 97.4%, 95.1% and 96.9%, 97.1%, 96.6%, 96.1%, 94.7%. CONCLUSION The improved method is more sensitive and reproducible, and can be used for quality control of the crude drug and preparation of phenylbutazone efficiently.