酒苁蓉HPLC特征图谱及5个苯乙醇苷类成分含量测定
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摘要
目的:采用高效液相色谱法建立酒苁蓉的特征图谱,并同时测定5个苯乙醇苷类成分的含量。方法:以松果菊苷、肉苁蓉苷A、毛蕊花糖苷、异毛蕊花糖苷和2′-乙酰基毛蕊花糖苷为对照,采用Agilent EclipseXDB-C_(18)(4.6 mm×250 mm,5μm)色谱柱,以甲醇为流动相A,0.1%甲酸为流动相B,梯度洗脱(0～15min,15%A→21%A;15～20 min,21%A→25%A;20～70 min,25%A→43%A),流速1.0 mL·min~(-1),检测波长为330 nm,柱温35℃。结果:建立酒苁蓉特征图谱,70 min内酒苁蓉的主要色谱峰能够达到完全分离,同时标定5个苯乙醇苷类成分,对15批饮片中的5个成分进行含量测定。松果菊苷、肉苁蓉苷A、毛蕊花糖苷、异毛蕊花糖和2′-乙酰基毛蕊花糖苷的质量浓度分别在10.05～1 005μg·mL~(-1)(r=0.999 9,n=6)、2.02～202μg·mL~(-1)(r=0.999 9,n=6)、2.04～204μg·mL~(-1)(r=0.999 9,n=6)、2.01～201μg·mL~(-1)(r=0.999 9,n=6)、2.02～202μg·mL~(-1)(r=0.999 9,n=6)范围内线性关系良好,方法的平均回收率(n=6)分别为99.8%(RSD=0.30%)、99.5%(RSD=1.6%)、100.2%(RSD=0.44%)、99.5%(RSD=0.53%)、99.8%(RSD=0.36%);15批酒苁蓉中上述5个成分含量分别为0.20%～7.38%、0.03%～0.20%、0.03%～0.72%、0.03%～0.75%、0.03%～0.07%。结论:该方法操作简单,重复性好,并能同时测定5个苯乙醇苷类成分含量,具备定性定量双重作用,可为酒苁蓉的质量标准研究提供更加全面的依据。
Objective:To establish an HPLC characteristic chromatogram and quantitative method for five components in Cistanches Herba. Methods:With echinacoside,cistanoside A,acteoside,iso-acteoside and 2′-acetyacteoside as reference substances,HPLC analysis was performed on an Agilent Eclipse XDB-C_(18) column(4.6 mm×250 mm,5 μm). The mobile phase was methanol-0.1% formic acid solution with gradient elution(0-15 min,15%A → 21%A;15-20 min,21%A → 25%A;20-70 min,25%A → 43%A). The flow rate was 1 mL·min~(-1),the detection wavelength was 330 nm and the column temperature was 35 ℃.Results:HPLC characteristic chromatogram of Cistanches Herba was established and five characteristic peaks were marked.Five characteristic peaks were simultaneously determined by HPLC within 70 min. The linear ranges of echinacoside,cistanoside A,acteoside,iso-acteoside and 2 ′-acetyacteoside were 10.05-1005 μg·mL~(-1)(r=0.999 9,n=6),2.02-202 μg·mL~(-1)(r=0.999 9,n=6),2.04-204 μg·mL~(-1)(r=0.999 9,n=6),2.01-201 μg·mL~(-1)(r=0.999 9,n=6)and 2.02-202 μg·mL~(-1)(r=0.999 9,n=6),respectively.The average recovery rate(n=6)were 99.8%(RSD=0.30%),99.5%(RSD=1.6%),100.2%(RSD=0.44%),99.5%(RSD=0.53%)and 99.8%(RSD=0.36%),respectively.The contents of the five components in 15 batches of Cistanches Herba were 0.20%-7.39%,0.03%-0.20%,0.03%-0.72%,0.03%-0.75% and 0.03%-0.07%,respectively.Conclusion:The proposed specific HPLC characteristic chromatogram and quantitation method of five components for Cistanches Herba is simple and repeatable,which offer reference for quality control of the drug.
Objective:To establish an HPLC characteristic chromatogram and quantitative method for five components in Cistanches Herba. Methods:With echinacoside,cistanoside A,acteoside,iso-acteoside and 2′-acetyacteoside as reference substances,HPLC analysis was performed on an Agilent Eclipse XDB-C_(18) column(4.6 mm×250 mm,5 μm). The mobile phase was methanol-0.1% formic acid solution with gradient elution(0-15 min,15%A → 21%A;15-20 min,21%A → 25%A;20-70 min,25%A → 43%A). The flow rate was 1 mL·min~(-1),the detection wavelength was 330 nm and the column temperature was 35 ℃.Results:HPLC characteristic chromatogram of Cistanches Herba was established and five characteristic peaks were marked.Five characteristic peaks were simultaneously determined by HPLC within 70 min. The linear ranges of echinacoside,cistanoside A,acteoside,iso-acteoside and 2 ′-acetyacteoside were 10.05-1005 μg·mL~(-1)(r=0.999 9,n=6),2.02-202 μg·mL~(-1)(r=0.999 9,n=6),2.04-204 μg·mL~(-1)(r=0.999 9,n=6),2.01-201 μg·mL~(-1)(r=0.999 9,n=6)and 2.02-202 μg·mL~(-1)(r=0.999 9,n=6),respectively.The average recovery rate(n=6)were 99.8%(RSD=0.30%),99.5%(RSD=1.6%),100.2%(RSD=0.44%),99.5%(RSD=0.53%)and 99.8%(RSD=0.36%),respectively.The contents of the five components in 15 batches of Cistanches Herba were 0.20%-7.39%,0.03%-0.20%,0.03%-0.72%,0.03%-0.75% and 0.03%-0.07%,respectively.Conclusion:The proposed specific HPLC characteristic chromatogram and quantitation method of five components for Cistanches Herba is simple and repeatable,which offer reference for quality control of the drug.
引文
[1]中华人民共和国药典2015年版.一部[S].2015:216ChP 2015.VolⅠ[S].2015:216
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[7]李承花,刘婧慧,徐珍霞,等.RP-HPLC同时测定管花肉苁蓉中4种苯乙醇苷的含量[J].药物分析杂志,2010,30(6):1003LICH,LIUJH,XUZX,etal.RP-HPLC simultaneous determination of four phenylethanoid glycosides in Cistanche tubulosa(Schrenk)[J].Chin J Pharm Anal,2010,30(6):1003
[8]彭芳,徐荣,王夏,等.不同等级整株肉苁蓉的药材品质和产地加工蒸制时间研究[J].中药材,2016,39(2):302PENG F,XU R,WANG X,et al.Effect of grade and steaming time on quality of cultivated Cistanche deserticola[J].J Chin Med Mater,2016,39(2):302
[9]马志国,谭咏欣.酒蒸不同时间肉苁蓉中6种苯乙醇苷类成分的变化[J].中成药,2011,33(11):1951MA ZG,TAN YX.Contents changes of six phenylethanoid glycosides under steaming time spans with wine in desertliving Cistanche[J].Chin Tradit Pat Med,2011,33(11):1951
[10]钱浩,喻芳君,耿宗成,等.不同季节采收的肉苁蓉中4种苯乙醇苷的含量比较研究[J].药物分析杂志,2016,36(11):1971QIAN H,YU FJ,GENG ZC,et al.Comparative study of the content of four phenylethanoid glycosides in Cistanche deserticola Y.C.Ma from different seasons[J].Chin J Pharm Anal,2016,36(11):1971
[2]雷丽,宋志宏,屠鹏飞.肉苁蓉属植物的化学成分研究进展[J].中草药,2003,34(5):473LEI L,SONG ZH,TU PF.Advances in research of chemical constituents in plants of Cistanche Hoffing.et Link.[J].Chin Tradit Herb Drugs,2003,34(5):473
[3]曹振杰,赵文军,吴雪萍.肉苁蓉中苯乙醇甙类化合物的LC-MS分析[J].中药材,2004,27(3):175CAO ZJ,ZHAO WJ,WU XP.Determination of phenylethanoid glycosides from Cistanche deserticola in spring and autumn with LC-MS[J].J Chin Med Mater,2004,27(3):175
[4]宋志宏,雷丽,屠鹏飞.肉苁蓉属植物的药理活性研究进展[J].中草药,2003,34(9):113SONG ZH,LEI L,TU PF.Advances in research of pharmacological activity in plants of Cistanche Hoffing.et Link.[J].Chin Tradit Herb Drugs,2003,34(9):113
[5]马志国.沙苁蓉与肉苁蓉饮片HPLC指纹图谱鉴别研究[J].中国药学杂志,2011,46(12):899MA ZG.Differentiation of processed Cistanche sinensis G.Beck and Herba Cistanches by HPLC[J].Chin Pharm J,2011,46(12):899
[6]朱乃亮,徐荣,吴海峰,等.荒漠肉苁蓉和管花肉苁蓉指纹图谱比较研究[J].中国药学杂志,2016,51(13):1116ZHU NL,XU R,WU HF,et al.Fingerprint comparative analysis of Cistanche deserticola Y.C.Ma and Cistanche tubulosa(Schrenk)Wight[J].Chin Pharm J,2016,51(13):1116
[7]李承花,刘婧慧,徐珍霞,等.RP-HPLC同时测定管花肉苁蓉中4种苯乙醇苷的含量[J].药物分析杂志,2010,30(6):1003LICH,LIUJH,XUZX,etal.RP-HPLC simultaneous determination of four phenylethanoid glycosides in Cistanche tubulosa(Schrenk)[J].Chin J Pharm Anal,2010,30(6):1003
[8]彭芳,徐荣,王夏,等.不同等级整株肉苁蓉的药材品质和产地加工蒸制时间研究[J].中药材,2016,39(2):302PENG F,XU R,WANG X,et al.Effect of grade and steaming time on quality of cultivated Cistanche deserticola[J].J Chin Med Mater,2016,39(2):302
[9]马志国,谭咏欣.酒蒸不同时间肉苁蓉中6种苯乙醇苷类成分的变化[J].中成药,2011,33(11):1951MA ZG,TAN YX.Contents changes of six phenylethanoid glycosides under steaming time spans with wine in desertliving Cistanche[J].Chin Tradit Pat Med,2011,33(11):1951
[10]钱浩,喻芳君,耿宗成,等.不同季节采收的肉苁蓉中4种苯乙醇苷的含量比较研究[J].药物分析杂志,2016,36(11):1971QIAN H,YU FJ,GENG ZC,et al.Comparative study of the content of four phenylethanoid glycosides in Cistanche deserticola Y.C.Ma from different seasons[J].Chin J Pharm Anal,2016,36(11):1971