蔗糖及其有关物质含量的HPLC-CAD分析研究
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摘要
目的:建立蔗糖及其有关物质含量的HPLC-CAD分析测定法。方法:采用Waters XBridge Amide色谱柱(4.6 mm×250 mm,3.5μm),柱温为30℃;以乙腈-0.2%三乙胺水溶液(65∶35)为流动相,流速为1.0 mL·min~(-1);检测器为电喷雾检测器(CAD),雾化温度HIGH(50℃),幂率1.15,采样频率10 Hz,过滤常数5.0 s。结果:蔗糖与相邻杂质以及各杂质峰之间均达到基线分离;蔗糖质量浓度在4.95～99.08μg·mL~(-1)及0.30～0.80 mg·mL~(-1)范围内线性关系良好(r>0.999),可分别用于有关物质检查和含量测定;检测下限为1.49μg·mL~(-1),定量下限为4.95μg·mL~(-1);进样精密度RSD为0.31%;重复性RSD为0.61%。4个批次的蔗糖样品含量均在99.09%～100.48%之间,有关物质含量在0.15%～0.25%之间。结论:所建方法专属、灵敏、耐用,操作简单,具有很好的应用前景。
Objective:To establish an HPLC-CAD method for the determination of sucrose and its related substances. Methods:The determination was carried out on a Waters XBridge Amide(4.6 mm×250 mm,3.5 μm) column at 30 ℃,with acetonitrile-0.2% triethylamine solution(65∶35)as mobile phase at a flow rate of 1.0 mL·min~(-1).The detector of charged aerosol detector(CAD)was applied with at high nebulization temperature at50 ℃ and collection frequency at 10 Hz with filter constant of 3.6 s,and PFV of 1.15.Results:Under the above chromatographic conditions,this method successfully achieved baseline separation among the peaks of sucrose,it's adjacent impurities and the other impurities obtained.The fine linear ranges of sucrose were between 4.95-99.08 μg·mL~(-1) and 0.30-0.80 mg·mL~(-1) with the correlation coefficients>0.999,suitable to the test of related substances and assay of sucrose,respectively.The LOD was 1.49 μg·m L~(-1) and the LOQ was 4.95 μg·m L~(-1).The RSDs of precision and repeatability were 0.31% and 0.61%,respectively.The contents of sucrose in four batches were all in the ranges of 99.09%-100.48%,and the total impurity was all in the ranges of 0.15%-0.25%.Conclusion:The established HPLC-CAD method is specific,sensitive,robust,and easy to operate with a promising prospect.
Objective:To establish an HPLC-CAD method for the determination of sucrose and its related substances. Methods:The determination was carried out on a Waters XBridge Amide(4.6 mm×250 mm,3.5 μm) column at 30 ℃,with acetonitrile-0.2% triethylamine solution(65∶35)as mobile phase at a flow rate of 1.0 mL·min~(-1).The detector of charged aerosol detector(CAD)was applied with at high nebulization temperature at50 ℃ and collection frequency at 10 Hz with filter constant of 3.6 s,and PFV of 1.15.Results:Under the above chromatographic conditions,this method successfully achieved baseline separation among the peaks of sucrose,it's adjacent impurities and the other impurities obtained.The fine linear ranges of sucrose were between 4.95-99.08 μg·mL~(-1) and 0.30-0.80 mg·mL~(-1) with the correlation coefficients>0.999,suitable to the test of related substances and assay of sucrose,respectively.The LOD was 1.49 μg·m L~(-1) and the LOQ was 4.95 μg·m L~(-1).The RSDs of precision and repeatability were 0.31% and 0.61%,respectively.The contents of sucrose in four batches were all in the ranges of 99.09%-100.48%,and the total impurity was all in the ranges of 0.15%-0.25%.Conclusion:The established HPLC-CAD method is specific,sensitive,robust,and easy to operate with a promising prospect.
引文
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[2]帅放文,章家伟,王向峰,等.HPLC-ELSD法测定药用辅料蔗糖的含量[J].中国药品标准,2014,15(6):446SHUAI FW,ZHANG JW,WANG XF,et al.HPLC-ELSDdetermination of excipients sucrose[J].Drug Stand China,2014,15(6):446
[3]中华人民共和国药典2015年版.四部[S].2015:631ChP 2015.VolⅣ[S].2015:631
[4]EP 9.0[S].2017:3664,2560
[5]USP 41-NF 36[S].2018:5626,1191
[6]JP 17[S].2017:1611
[7]田春霞,石文宏.高效液相色谱法测定药用辅料蔗糖的含量[J].中国医药指南,2012,10(24):71TIAN CX,SHI WH.Determination of sucrose in pharmaceutical excipients by high performance liquid chromatography[J].Guide China Med,2012,10(24):71
[8]郑慧,武世敏,李宏军.挤压玉米淀粉做辅料时啤酒麦汁中的可发酵糖组分分析[J].湖北农业科学,2015,54(1):178ZHENG H,WU SM,LI HJ.Analyzing the components of fermentable sugar in beer wort using extruded maize starch as adjunt[J].Hubei Agric Sci,2015,54(1):178
[9]LV GP,HU DJ,CHEONG KL,et al.Decoding glycome of Astragalus membranaceus based on pressurized liquid extraction,microwave-assisted hydrolysis and chromatographic analysis[J].JChromatogr A,2015,1409:19
[10]YU L,LI X K,ZHANG HF,et al.Simultaneous determination of monosaccharide and oligosaccharides by RP-HPLC-RID in Huai-Radix Rehmanniae[J].Chin J Pharm Anal,2013,33(6):977
[11]LINDQVIST DN,PEDERSEN H-E,RASMUSSEN LH.A novel technique for determination of the fructose,glucose and sucrose distribution in nectar from orchids by HPLC-ELSD[J].JChromatogra B,2018,1081-1082:126
[12]LI FJ,ZHANG HY,LI Y,et al.Simultaneous identification and quantification of dextran 20 and sucrose in lyophilized thrombin powder by size exclusion chromatography with ELSD[J].Chromatographia,2012,75(3):187
[13]邹学权,傅飞霞,陈孜.离子色谱-脉冲安培法同时测定婴幼儿配方食品中的蔗糖与乳糖[J].浙江化工,2018,49(3):48ZOU XQ,FU FX,CHEN Z.Determination of sucrose and lactose in infant formula food by ion chromatography[J].Zhejiang Chem Ind,2018,49(3):48
[14]GEORGELIS N,FENCIL K,RICHAEL CM.Validation of a rapid and sensitive HPLC/MS method for measuring sucrose,fructose and glucose in plant tissues[J].Food Chem,2018,262:191
[15]LIU LY,LIU X.An innovative universal detector-corona charged aerosol detector[J].Mod Sci Instrum,2011,5(10):141
[16]刘路,高旋,杨永健.HPLC-电雾式检测器的应用[J].中国医药工业杂志,2012,43(3):227LIU L,GAO X,YANG YJ.Application of HPLC-charged aerosol detector[J].Chin J Pharm,2012,43(3):227
[17]孙宝良,李百芳,王建祺,等.用测定比旋度的物理学方法来分析D-葡萄糖的变旋光现象[J].中国医学物理学杂志,2007,24(1):46SUN BL,LI BF,WANG JQ,et al.Analyzing mutarotation of D-glucose by physics ways determining specific rotation[J].Chin JMed Phys,2007,24(1):46
[18]赵永潜,张亚萍,许广建,等.D-葡萄糖变旋特性的实验研究[J].物理与工程,2016,26(6):94ZHAO YQ,ZHANG YP,XU GJ,et al.Research on experiment of the mutarotation of D-glucose[J].Phys Eng,2016,26(6):94
[19]EP 9.4[S].2018:5448
[20]国家药典委员会.蔗糖公示稿[EB/OL].(2018-10-15)[2018-10-15].http://www.chp.org.cn/view/ff80808165a20a8901667668019c14a8?a=BZFULIAOChinese Pharmacopoeia Commission.Draft Monograph of Sucrose for Public Comments[EB/OL].(2018-10-15)[2018-10-15].http://www.chp.org.cn/view/ff80808165a20a8901667668019c14a8?a=BZFULIAO