HPLC法测定赖氨酸维B_(12)颗粒中维生素B_(12)的含量及含量均匀度

Determination of the Content and Content Uniformity of Vitamin B_(12) in Lysine and Vitamin B_(12) Granules by HPLC

目的建立HPLC方法测定赖氨酸维B_(12)颗粒中维生素B_(12)含量及含量均匀度。方法采用高效液相色谱法,紫外检测器,以C_(18)色谱柱为分离柱,以甲醇-0.028 mol·L^(-1)磷酸氢二钠溶液(磷酸调节pH值至3.5)(20∶80)为流动相,在检测波长361 nm处测定维生素B_(12)含量。流速为1.0 mL·min^(-1),进样体积为100μL。结果维生素B_(12)浓度在0.2~5.0μg·mL-1范围内与峰面积呈良好的线性关系,平均回收率为105.80%(n=9)。按此方法分析了3个厂家5个批次样品,样品中维生素B_(12)的测定值为标示量的43.8%~93.3%,同时进行了含量均匀度评价试验。结果显示,某些批次样品中的维生素B_(12)存在含量偏低、含量均匀度结果不符合规定的现象,表明厂家的生产工艺可能存在一定的问题。结论本法结果准确可靠,重现性好,可作为赖氨酸维B_(12)颗粒中维生素B_(12)含量及含量均匀度测定方法。

OBJECTIVE To establish an HPLC method for the determination of the content and content uniformity of Vitamin B_(12) in Lysine and Vitamin B_(12) Granules.METHODS The HPLC method was conducted on a C_(18) column.The mobile phase was methanol-0.028 mol·L^(-1) disodium hydrogen phosphate solution(20∶80).The flow rate was 1.0 mL·min^(-1) and the detection wavelength was 361 nm.The injection volume was 100μL.RESULTS The linearity of Vitamin B_(12) was good within the range of 0.2-5.0μg·mL-1.The average recovery was 105.80%([WTBX]n[WTBZ]=9).Five batches of samples were analyzed in this manufacturers,the measured value of Vitamin B_(12) in the sample was 43.8%to 93.3%of the indicated amount,the content uniformity evaluation test was also carried out.The results showed that the content of Vitamin B_(12) in some batches of samples was low and the content uniformity results did not meet the requirements,indicating that the production process of the manufacturer may have some problems.CONCLUSION The method is accurate,reliable and reproducible,and can be used in the determination of content and content uniformity of Vitamin B_(12) in Lysine and Vitamin B_(12) Granules.