Abstract
Objective To establish a high-performance liquid chromatography(HPLC)method for the simultaneous determination of six components in Xiao'er Siwei Pugai Tablets.Methods The contents of glucose,sucrose,and vitamin D2 in the preparation were determined by the HPLC method,while the contents of vitamin B1,vitamin B2,and vitamin C in the preparation were determined by the ultra-high-performance liquid chromatography(UPLC)method.Chromatographic columns:the Ultimate XB-NH2 amino column(250 mm×4.6 mm,5 μm)was used for glucose and sucrose,the Acquity UPLC HSS T3 column(150 mm×2.1 mm,1.8 μm)was used for vitamin B1,vitamin B2 and vitamin C,and the Guangzhou Yanchuang Gensial® C18 column(150 mm×4.6 mm,5 μm)was used for vitamin D2.Mobile phases:acetonitrile-water-ammonia solution(80∶20∶0.1,V/V/V)was set for glucose and sucrose,0.02 mol/L ammonium dihydrogen phosphate solution(containing 0.05%sodium heptane sulfonate,adjusted to pH 3.0 with phosphoric acid)-methanol(80∶20,V/V)was set for vitamin B1,vitamin B2,and vitamin C,and acetonitrile-0.5%formic acid solution(95∶5,V/V)was set for vitamin D2.Flow rates:1.0 mL/min was set for glucose,sucrose,and vitamin D2,and 0.2 mL/min was set for vitamin B1,vitamin B2,and vitamin C.Detection wavelengths:267 nm was set for vitamin B1,vitamin B2,and vitamin C,and 264 nm was set for vitamin D2.Detectors:the UV-visible spectrophotometric refractive index detector was used for determining glucose and sucrose,the photo-diode array(PDA)detector was used for determining vitamin B1,vitamin B2,and vitamin C,and the UV-visible spectrophotometric detector was used for determining vitamin D2.Column temperatures:40℃was set for glucose,sucrose,and vitamin D2,and 35℃was set for vitamin B1,vitamin B2,and vitamin C.Injection volumes:10 μL was set for glucose,sucrose,and vitamin D2,and 1 μL was set for vitamin B1,vitamin B2,and vitamin C.Results The linear ranges for glucose,sucrose,vitamin B1,vitamin B2,vitamin C,and vitamin D2 were 1.00-20.00 mg/mL,1.00-20.00 mg/mL,0.80-24.00 μg/mL,0.40-12.00 μg/mL,10.00-300.00 μg/mL,and 0.10-10.00 μg/mL(R²≥0.999 1),respectively.The limits of detection of the above six components were 1.00 μg,2.00 μg,0.02 ng,0.01 ng,0.025 ng,and 0.20 ng,and the limits of quantification of the above six components were 3.00 μg,6.00 μg,0.80 ng,0.40 ng,0.10 ng,and 0.50 ng,respectively.The RSDs of the precision,stability,and repeatability tests were all lower than 2.50%.The average recoveries of the above six components were 99.18%,99.47%,98.83%,98.54%,99.49%,and 98.80%,with RSDs of 1.61%,2.28%,1.82%,2.47%,2.33%,and 2.72%(n=6),respectively.In the three batches of samples,the six components in each tablet were in the ranges of 148.702-153.218 mg,148.623-151.419 mg,0.198-0.203 mg,0.098-0.103 mg,2.488-2.512 mg,and 1.632-1.689 μg,respectively.Conclusion The method is simple,specific,reproducible,and accurate,which can be used for simultaneous determination of six components in Xiao'er Siwei Pugai Tablets.关键词
小儿四维葡钙片/高效液相色谱法/超高效液相色谱法/葡萄糖/蔗糖/维生素B1/维生素B2/维生素C/维生素D2/含量测定Key words
Xiao'er Siwei Pugai Tablets/HPLC/UPLC/glucose/sucrose/vitamin B1/vitamin B2/vitamin C/vitamin D2/content determination分类
医药卫生
