快速分离液相色谱法测定庆大霉素普鲁卡因维B_(12)颗粒中维生素B_(12)的含量

Determination of vitamin B_(12) in Gentamicin Sulfate,Procaine Hydrochloride and Vitamin B_(12) Granules by rapid resolution liquid chromatography

目的建立快速分离液相色谱(RRLC)法测定庆大霉素普鲁卡因维B12颗粒中维生素B12含量的方法。方法选用Agilent SB-C18柱(3.0 mm×150 mm,1.8μm);以0.005 mol·L-1磷酸二氢钾溶液(每1 000 mL加磷酸0.5 mL)-甲醇(75∶25)为流动相;流速为0.5 mL·min-1;柱温40℃;检测波长361 nm;进样体积200μL。结果 VB12在10~1 400 ng·mL-1浓度范围内与峰面积呈良好的线性关系(r=0.999 8,n=9)。平均加样回收率为100.2%(RSD为1.0%,n=6)。方法的最低检出限为8.0 ng·mL-1。供试品溶液色谱图中VB12峰与相邻峰的分离度良好,样品中其他组分不干扰VB12的测定。结论该方法快速,简便,重复性好,结果准确可靠,适用于庆大霉素普鲁卡因维B12颗粒中微量维生素B12的含量测定。

Objective To establish a RRLC method for the determination of trace VB^2 in Gentamicin Sulfate,Procaine Hydroehloride and Vitamin BI： Granules. Methods The chromatographic analysis were carried out on an Agilent SB - Cis Column（3.0 mmx 150 mm, 1.8μm） , and the mobile phase was O. 005 mol· L-1 potassium dihydrogen phosphate solution - methanol {75：25 ）. The detection was carried out at 361 nm,with a flow rate of 0. 5 mL· min-l. The column temperature was 40 ℃ and injection volume was 200 μL. Results VB12 emwes were linear in the range of 10 - 1 400 ng · mL -1 （ r = 0. 999 8, n = 9 ） , the average recovery was 100. 2% （ RSD was 1.0% , n = 6）. The LOD of the method was 8.0 ng· mL-1. The peak of VB12 had a good resolution to the other peaks in test solution,therefore the other components in sample had no interference to tile determination of VBI2. Conclusion This rapid and simple method showed high sensitivity. The results were accurate and reliable by RRLC method,which was suitable for the assay of trace VBI2 in Gentamicin Sulfate,Procaine Hydroehloride and Vitamin BI2 Granules.