UPLC-MS/ELSD法快速检测板蓝根颗粒及组分中的糖类成分  

Rapid determination of sugars in Banlangen Granule and its fractions by UPLC-MS/ELSD

作　　者：钱秀玉 聂黎行[1] 戴忠[1] 马双成[1] QIAN Xiuyu;NIE Lixing;DAI Zhong;MA Shuangcheng(National Institutes for Food and Drug Control,Beijing 100050,China)

机构地区：[1]中国食品药品检定研究院,北京100050

出　　处：《药物评价研究》2020年第7期1267-1272,共6页Drug Evaluation Research

基　　金：国家自然科学基金(81303194);重大新药创制专项课题(2018ZX09735006);中国食品药品检定研究院学科带头人培养基金(2017X1)。

摘　　要：目的建立板蓝根颗粒及组分中糖类成分的快速定性方法和主要单糖的定量测定方法。方法采用超高效液相色谱结合质谱检测器(UPLC-MS)快速定性板蓝根颗粒及组分中的糖类成分,采用超高效液相色谱结合蒸发光散射检测器(UPLC-ELSD)同时测定果糖、葡萄糖的含量。使用ACQUITY UPLC BEH Amide(2.1 mm×150 mm,1.7μm)色谱柱分离,柱温35℃。流动相分别为乙腈-0.1%氨水(含10 mmol/L碳酸氢铵)(75∶25)和乙腈(含0.2%三乙胺)-水(75∶25),体积流量0.2 mL/min。质谱检测器采用负离子模式,毛细管电压0.8V,锥孔电压4V,探头温度600℃,采集频率1 Hz,采用全扫描及SIR扫描模式。ELSD检测器漂移管温度50℃,氮气压力275.8 kPa。结果经UPLC-MS法鉴定板蓝根颗粒中糖类成分种类为单糖和双糖,主要为果糖、葡萄糖、蔗糖、肌醇,水洗脱组分检出果糖、葡萄糖、蔗糖,10%醇洗脱组分和20%醇洗脱组分均检出蔗糖。UPLC-ELSD法含量测定结果表明,果糖、葡萄糖在各自范围内线性关系良好,各相关系数均0.9993在以上,平均回收率分别为99.0%、100.1%。28个生产企业的28批样品糖类成分含量差异较大。结论所建立的方法准确度、重复性良好,具有操作简便、节省时间等特点,可为板蓝根颗粒的质量控制和物质基础研究提供参考。Objective A rapid qualitative method for the Sugars and a quantitation method for the main monosaccharides in Banlangen Granule and its fractions were established.Methods Sugars were determined by ultra performance liquid chromatography(UPLC)method combined with mass spectrometric detector(MS).Fructose and glucose were determined by UPLC method combined with evaporative light scattering detector(ELSD).Separation was performed on an ACQUITY UPLC BEH Amide(2.1 mm×150 mm,1.7μm)column and the column temperature was 35℃.The mobile phase consisted of acetonitrile(containing 0.2%triethylamine)-water(75:25)and acetonitrile-water(containing 0.1%ammonia and 10 mmol/L NH4HCO3)(75:25)at a flow rate of 0.2 mL/min.The MS spectra were acquired in negative mode with a capillary voltage of 0.8 kV,a cone voltage of 4.0 kV,a probe temperature of 600℃,an acquisition frequency of 1 Hz.The analytes were determined in full scan mode and SIR mode.The temperature for the evaporator tube of ELSD was maintained at 50℃and the gas pressure for nitrogen was 275.8 kPa.Results The types of Sugars in Banlangen Granule were monosaccharide and disaccharide.Fructose,glucose,sucrose and inositol in the sample of banlangen granules were detected by UPLC-MS method.Fructose,glucose and sucrose were detected in water fraction.Sucrose was detected in both 10%alcohol fraction and 20%alcohol fraction.The results of UPLC-ELSD method showed that the calibration curves of the 2 components showed good linearity within their test ranges.The correlation coefficients were not lower than 0.9993.The average recoveries for fructose,glucose were 99.0%,100.1%,respectively.There was a great variation among contents of sugars in 28 batches of samples from 28 manufactures.Conclusion The proposed method was accurate,repeatable,simple and fast,thus providing reference for quality control and basic substances research of Banlangen Granule.

关 键 词：板蓝根颗粒 果糖 葡萄糖 超高效液相色谱 蒸发光散射检测器 质谱检测器 

分 类 号：R286.02[医药卫生—中药学]