高效液相色谱法测定庆大霉素普鲁卡因维B12胶囊中维生素B12的含量及含量均匀度的评价

HPLC Determination of Vitamin B in Gentamicin Sulfate,Procaine Hydrochloride and Vitamin B Capsules and Evaluation of Content Uniformity

提出了高效液相色谱法测定A厂、B厂、C厂和D厂等4个厂家47个批次庆大霉素普鲁卡因维B12胶囊样品中维生素B12的含量,并对4个厂家相同批次或不同批次的样品中维生素B12含量的均匀度进行了评价。用水提取胶囊内容物中的维生素B12,以CAPCELL PAK ACR C_(18)色谱柱为分离柱,以不同比例混合的甲醇和0.028 mol·L^(-1)磷酸盐缓冲溶液(pH 3.2)为流动相进行梯度洗脱,在检测波长362 nm处测定目标物含量。结果显示:维生素B12标准曲线的线性范围为0.2~5.0 mg·L^(-1),测定下限(10S/N)为0.12 mg·L^(-1)。对胶囊样品进行3个浓度水平的加标回收试验,所得回收率为96.9%~102%。6份样品溶液中维生素B12测定值的相对标准偏差为1.5%。按此方法分析了47个批次样品,D厂样品中维生素B12的测定值为标示量的55.4%~137.0%,超出了根据中国药典二部和实际样品设定的范围(80.0%~120.0%)。故又进行了含量均匀度评价试验,结果显示,D厂批次内和批次间样品中的维生素B12均存在含量不均匀的现象,查找原因发现,这可能与D厂使用的干法混合工艺有关。

A HPLC method was introduced to determine the content of Vitamin B12 in 47 batches of gentamicin sulfate,procaine hydrochloride and Vitamin B12 capsules from 4 manufacturers including factory A,factory B,factory C,factory D,and the content uniformity was evaluated for Vitamin B12 in the same batch or different bathes of samples from 4 manufactures.Vitamin B12 in capsules was extracted with water.CAPCELL PAK ACR C_(18) chromatographic column was used as separation column,the mixtures of methanol and 0.028 mol·L^(-1)sodium dihydrogen phosphate solution(pH 3.2)with various volume ratios were used as mobile phase for gradient elution,and the content of Vitamin B12 was determined at the detection wavelength of 362 nm.The results showed that the linearity range of Vitamin B12 standard curve was in the range of 0.2-5.0 mg·L^(-1),with lower limit of determination(10 S/N)of 0.12 mg·L^(-1).The spiked recovery was made on capsule samples at three concentration levels,giving recoveries in the range of 96.9%-102%,and RSD of measured values of Vitamin B12 in 6 sample solutions was 1.5%.This method was used to analyze the 47 batches of samples,and values of Vitamin B12 obtained were in the range of 55.4%-137.0%of the labelled values of the samples from factory D,exceeding the range(80.0%-120.0%)set by the second part of Chinese Pharmacopoeia and actual samples.Therefore,the content uniformity evaluation was made,and the results showed that the vitomin B12 content in the samples within and between batches was uneven,finding that it may be related to the dry mixing process used in factory D.