在线二维柱切换高效液相色谱法同时测定牙膏中三七皂苷R1、人参皂苷Rg1、Re和Rb1
张岩等
摘 要 使用双梯度液相色谱系统紫外检测器，建立了二维液相色谱法全自动快速同时测定牙膏中三七皂苷R1、人参皂苷Rg1、Re和Rb1的含量。样品经超声提取后，以Syncronis C18为一维分析柱，ODS C18为二维分析柱，利用一维色谱柱完成三七皂苷R1和人参皂苷Rb1分离测定以及人参皂苷Rg1和人参皂苷Re的净化； 利用二维色谱柱完成人参皂苷Rg1和人参皂苷Re的分析。一维分析和二维分析均以乙腈水体系作为流动相，梯度洗脱，检测波长为203 nm，整个分析过程仅需30 min。三七皂苷R1、人参皂苷Rg1、Re和Rb1在0.5～200 mg/L范围内线性良好，相关系数R2分别为0.9994， 0.9996， 0.9995和0.9994，平均回收率均在86.4%～95.1%之间。本方法简便快速，测定结果准确可靠，可用于牙膏中三七皂苷R1、人参皂苷Rg1、Re和Rb1含量的测定。
3.3 一维流动相的选择
考察了流动相为乙腈水体系和甲醇水体系对一维
色谱保留时间及峰形的影响，结果表明，使用甲醇水为流动相三七皂苷R1、人参皂苷Rg1和Re三者不能达到基线分离，且峰形较宽（图4）。无法确定人参皂苷Rg1和Re切入二维色谱柱的时间，因色谱峰形过宽，需要切入时间长，造成流动相比例不匹配，容易引起压力波动，从而影响到二维色谱中目标物保留时间的漂移。采用乙腈水作为一维色谱流动相时，虽然人参皂苷Rg1和Re重合在一起，但二者能够与三七皂苷R1达到基线分离分析，且峰形良好，峰窄，从而确定了二维分析的切入时间，故选择了乙腈水体系作为一维流动相（图4）。通过优化实验条件发现乙腈水体系应用于二维色谱能够将人参皂苷Rg1和Re分离，为了使流动相匹配，避免压力波动，所以二维流动相选择乙腈水体系。
在一维分析中三七皂苷R1和人参皂苷Rb1较容易分离，所以在一维色谱中采用高比例的有机相和细粒径的色谱柱，提高分析速度，使在一维色谱中较难分离且存在基质干扰的人参皂苷Rg1和Re切换到二维色谱柱中继续分离，常规色谱分析4种物质需60 min以上，本方法仅需30 min，提高了分析速度，同时解决了人参皂苷Rg1和Re存在干扰和分离的难题。
实验结果表明， 通过在线二维柱切换，双梯度洗脱，简化了分析过程，提高了分析速度，有效去除了复杂基质的干扰。本方法可作为牙膏中三七皂苷R1、人参皂苷Rg1， Re和Rb1的常规检测方法，为监管部门规范管理口腔卫生产品市场提供了技术支撑。
References
1 WANG Qiang， JIANG YingQiao， MA ShiPing， DANG XueDong. China Journal of Chinese Materia Medica， 2000， 25（10）： 617-618
王 强， 江英桥， 马世平， 党学东. 中国中药杂志， 2000， 25（10）： 617-618
2 ZHANG XiaoPing， XU ShiJie. Chin. JMAP， 2006， 23（3）： 219-220
张小平， 徐世杰. 中国现代应用药学杂志， 2006， 23（3）： 219-220
3 Li L， Zhang J L， Sheng Y X， Guo D A， Wang Q， Guo H Z. J. Pharm. Biome. Anal.， 2005， 38（1）： 45-51
4 Lau A J， Woo S O， Koh H L. J. Chromatogr. A， 2003， 1011（12）： 77-87
5 XUE YiBo， GUO XiaoHuan. China Medicine and Pharmacy， 2012， 2（18）： 105-108
薛毅博， 郭晓焕. 中国医药科学， 2012， 2（18）： 105-108
6 Dong T T， Cui X M， Song Z H， Zhao K J， Ji Z N， Lo C K， Tsim K W K. J. Agric. Food Chem.， 2003， 51（16）： 4617-4623
7 Shangguan D H， Han H W， Zhao R， Zhao Y X， Xiong S X， Liu G Q. J. Chromatogr. A， 2001， 910（2）： 367-372
8 Li W， Fitzloff J F. J. Pharm. Pharmacol.， 2001， 53（12）： 1637-1643
9 SHA DongXu， ZHANG ManLai. Chin. J. Chin. Mater Med.， 2005， 30（2）： 112-115
沙东旭， 张满来. 中国药杂志， 2005， 30（2）： 112-115
10 Huang Y Z， Wang N S. Trad. Chin. Drug Res. Clin. Pharmacol.， 2003， 14， 180-182
11 Wan J B， Li P， Li S P， Wang Y， Dong T T， Tsim K W. J. Sep. Sci.， 2006， 29（14）： 2190-2196
12 Li L， Zhang J L， Sheng Y X， Ye G， Guo H Z， Guo D A. J. Chromatogr. B， 2004， 808（2）： 177-183
13 Cui M， Song F， Zhou Y， Liu Z， Liu S. Rapid Commun. Mass Spectrom.， 2000， 14（14）： 1280-1286
14 Zu Y G， Yan M M， Fu Y J， Liu W， Zhang L， Gu C B， Efferth T. J. Sep. Sci.， 2009， 32（4）： 517-525
15 Bompadre S， Tagliabracci A， Battino M， Giorgetti R. J. Chromatogr. B， 2008， 863（1）： 177-180
Simultaneous Quantification of Notoginsenoside R1 and Ginsenoside
Rg1， Re， Rb1 in Tooth Paste by Liquid Chromatography Coupled
with Fully Automated Online Twodimensional Column Switching Method
ZHANG Yan1，2， MA XiaoFei2， L Pin1， CONG Bin*1
1（Hebei Key Laboratory of Forensic Medicine， Department of Forensic Medicine，
Hebei Medical University， Shijiazhuang 050017， China）
2（Hebei Key Laboratory of Food Safety， Hebei Food Inspection and Research Institute， Shijiazhuang 050091， China）
Abstract A novel automated method for simultaneous quantification of notoginsenoside R1 and ginsenoside Rg1， Re， Rb1 in tooth paste was developed using twodimensional columnswitching chromatography with dual gradient liquid chromatography system coupled with UV detector. The determination of notoginsenoside R1 and ginsenoside Rb1 and the purification of ginsenoside Rg1， Re were accomplished in the onedimensional column. The separation of ginsenoside Rg1， Re was finished on the twodimensional column. Syncronis C18 was used as the onedimensional column with the mobile phase of acetonitrile and water， and ODS C18 was employed as the twodimensional column with the mobile phase was as same as in onedimensional column. The detection wavelength was set at 203 nm. The determination was completed in 30 min. The good linearities of notoginsenoside R1 and ginsenoside Rg1， Re， Rb1 were proved respectively in the range of 0.5-200 mg/L and the correlation coefficients （R2） were 0.9994， 0.9996， 0.9995 and 0.9994 respectively. The mean recoveries of notoginsenoside R1 and ginsenoside Rg1， Re， Rb1 were 86.4%-95.1%. It was proved that this method could greatly improve the efficiency of sample analysis.
Keywords High performance liquid chromatography； Twodimensional separation； Notoginsenoside； Ginsenoside
（Received 20 August 2014； accepted 7 October 2014
14 Zu Y G， Yan M M， Fu Y J， Liu W， Zhang L， Gu C B， Efferth T. J. Sep. Sci.， 2009， 32（4）： 517-525
15 Bompadre S， Tagliabracci A， Battino M， Giorgetti R. J. Chromatogr. B， 2008， 863（1）： 177-180
Simultaneous Quantification of Notoginsenoside R1 and Ginsenoside
Rg1， Re， Rb1 in Tooth Paste by Liquid Chromatography Coupled
with Fully Automated Online Twodimensional Column Switching Method
ZHANG Yan1，2， MA XiaoFei2， L Pin1， CONG Bin*1
1（Hebei Key Laboratory of Forensic Medicine， Department of Forensic Medicine，
Hebei Medical University， Shijiazhuang 050017， China）
2（Hebei Key Laboratory of Food Safety， Hebei Food Inspection and Research Institute， Shijiazhuang 050091， China）
Abstract A novel automated method for simultaneous quantification of notoginsenoside R1 and ginsenoside Rg1， Re， Rb1 in tooth paste was developed using twodimensional columnswitching chromatography with dual gradient liquid chromatography system coupled with UV detector. The determination of notoginsenoside R1 and ginsenoside Rb1 and the purification of ginsenoside Rg1， Re were accomplished in the onedimensional column. The separation of ginsenoside Rg1， Re was finished on the twodimensional column. Syncronis C18 was used as the onedimensional column with the mobile phase of acetonitrile and water， and ODS C18 was employed as the twodimensional column with the mobile phase was as same as in onedimensional column. The detection wavelength was set at 203 nm. The determination was completed in 30 min. The good linearities of notoginsenoside R1 and ginsenoside Rg1， Re， Rb1 were proved respectively in the range of 0.5-200 mg/L and the correlation coefficients （R2） were 0.9994， 0.9996， 0.9995 and 0.9994 respectively. The mean recoveries of notoginsenoside R1 and ginsenoside Rg1， Re， Rb1 were 86.4%-95.1%. It was proved that this method could greatly improve the efficiency of sample analysis.
Keywords High performance liquid chromatography； Twodimensional separation； Notoginsenoside； Ginsenoside
（Received 20 August 2014； accepted 7 October 2014
14 Zu Y G， Yan M M， Fu Y J， Liu W， Zhang L， Gu C B， Efferth T. J. Sep. Sci.， 2009， 32（4）： 517-525
15 Bompadre S， Tagliabracci A， Battino M， Giorgetti R. J. Chromatogr. B， 2008， 863（1）： 177-180
Simultaneous Quantification of Notoginsenoside R1 and Ginsenoside
Rg1， Re， Rb1 in Tooth Paste by Liquid Chromatography Coupled
with Fully Automated Online Twodimensional Column Switching Method
ZHANG Yan1，2， MA XiaoFei2， L Pin1， CONG Bin*1
1（Hebei Key Laboratory of Forensic Medicine， Department of Forensic Medicine，
Hebei Medical University， Shijiazhuang 050017， China）
2（Hebei Key Laboratory of Food Safety， Hebei Food Inspection and Research Institute， Shijiazhuang 050091， China）
Abstract A novel automated method for simultaneous quantification of notoginsenoside R1 and ginsenoside Rg1， Re， Rb1 in tooth paste was developed using twodimensional columnswitching chromatography with dual gradient liquid chromatography system coupled with UV detector. The determination of notoginsenoside R1 and ginsenoside Rb1 and the purification of ginsenoside Rg1， Re were accomplished in the onedimensional column. The separation of ginsenoside Rg1， Re was finished on the twodimensional column. Syncronis C18 was used as the onedimensional column with the mobile phase of acetonitrile and water， and ODS C18 was employed as the twodimensional column with the mobile phase was as same as in onedimensional column. The detection wavelength was set at 203 nm. The determination was completed in 30 min. The good linearities of notoginsenoside R1 and ginsenoside Rg1， Re， Rb1 were proved respectively in the range of 0.5-200 mg/L and the correlation coefficients （R2） were 0.9994， 0.9996， 0.9995 and 0.9994 respectively. The mean recoveries of notoginsenoside R1 and ginsenoside Rg1， Re， Rb1 were 86.4%-95.1%. It was proved that this method could greatly improve the efficiency of sample analysis.
Keywords High performance liquid chromatography； Twodimensional separation； Notoginsenoside； Ginsenoside
（Received 20 August 2014； accepted 7 October 2014
摘 要 使用双梯度液相色谱系统紫外检测器，建立了二维液相色谱法全自动快速同时测定牙膏中三七皂苷R1、人参皂苷Rg1、Re和Rb1的含量。样品经超声提取后，以Syncronis C18为一维分析柱，ODS C18为二维分析柱，利用一维色谱柱完成三七皂苷R1和人参皂苷Rb1分离测定以及人参皂苷Rg1和人参皂苷Re的净化； 利用二维色谱柱完成人参皂苷Rg1和人参皂苷Re的分析。一维分析和二维分析均以乙腈水体系作为流动相，梯度洗脱，检测波长为203 nm，整个分析过程仅需30 min。三七皂苷R1、人参皂苷Rg1、Re和Rb1在0.5～200 mg/L范围内线性良好，相关系数R2分别为0.9994， 0.9996， 0.9995和0.9994，平均回收率均在86.4%～95.1%之间。本方法简便快速，测定结果准确可靠，可用于牙膏中三七皂苷R1、人参皂苷Rg1、Re和Rb1含量的测定。
3.3 一维流动相的选择
考察了流动相为乙腈水体系和甲醇水体系对一维
色谱保留时间及峰形的影响，结果表明，使用甲醇水为流动相三七皂苷R1、人参皂苷Rg1和Re三者不能达到基线分离，且峰形较宽（图4）。无法确定人参皂苷Rg1和Re切入二维色谱柱的时间，因色谱峰形过宽，需要切入时间长，造成流动相比例不匹配，容易引起压力波动，从而影响到二维色谱中目标物保留时间的漂移。采用乙腈水作为一维色谱流动相时，虽然人参皂苷Rg1和Re重合在一起，但二者能够与三七皂苷R1达到基线分离分析，且峰形良好，峰窄，从而确定了二维分析的切入时间，故选择了乙腈水体系作为一维流动相（图4）。通过优化实验条件发现乙腈水体系应用于二维色谱能够将人参皂苷Rg1和Re分离，为了使流动相匹配，避免压力波动，所以二维流动相选择乙腈水体系。
在一维分析中三七皂苷R1和人参皂苷Rb1较容易分离，所以在一维色谱中采用高比例的有机相和细粒径的色谱柱，提高分析速度，使在一维色谱中较难分离且存在基质干扰的人参皂苷Rg1和Re切换到二维色谱柱中继续分离，常规色谱分析4种物质需60 min以上，本方法仅需30 min，提高了分析速度，同时解决了人参皂苷Rg1和Re存在干扰和分离的难题。
实验结果表明， 通过在线二维柱切换，双梯度洗脱，简化了分析过程，提高了分析速度，有效去除了复杂基质的干扰。本方法可作为牙膏中三七皂苷R1、人参皂苷Rg1， Re和Rb1的常规检测方法，为监管部门规范管理口腔卫生产品市场提供了技术支撑。
References
1 WANG Qiang， JIANG YingQiao， MA ShiPing， DANG XueDong. China Journal of Chinese Materia Medica， 2000， 25（10）： 617-618
王 强， 江英桥， 马世平， 党学东. 中国中药杂志， 2000， 25（10）： 617-618
2 ZHANG XiaoPing， XU ShiJie. Chin. JMAP， 2006， 23（3）： 219-220
张小平， 徐世杰. 中国现代应用药学杂志， 2006， 23（3）： 219-220
3 Li L， Zhang J L， Sheng Y X， Guo D A， Wang Q， Guo H Z. J. Pharm. Biome. Anal.， 2005， 38（1）： 45-51
4 Lau A J， Woo S O， Koh H L. J. Chromatogr. A， 2003， 1011（12）： 77-87
5 XUE YiBo， GUO XiaoHuan. China Medicine and Pharmacy， 2012， 2（18）： 105-108
薛毅博， 郭晓焕. 中国医药科学， 2012， 2（18）： 105-108
6 Dong T T， Cui X M， Song Z H， Zhao K J， Ji Z N， Lo C K， Tsim K W K. J. Agric. Food Chem.， 2003， 51（16）： 4617-4623
7 Shangguan D H， Han H W， Zhao R， Zhao Y X， Xiong S X， Liu G Q. J. Chromatogr. A， 2001， 910（2）： 367-372
8 Li W， Fitzloff J F. J. Pharm. Pharmacol.， 2001， 53（12）： 1637-1643
9 SHA DongXu， ZHANG ManLai. Chin. J. Chin. Mater Med.， 2005， 30（2）： 112-115
沙东旭， 张满来. 中国药杂志， 2005， 30（2）： 112-115
10 Huang Y Z， Wang N S. Trad. Chin. Drug Res. Clin. Pharmacol.， 2003， 14， 180-182
11 Wan J B， Li P， Li S P， Wang Y， Dong T T， Tsim K W. J. Sep. Sci.， 2006， 29（14）： 2190-2196
12 Li L， Zhang J L， Sheng Y X， Ye G， Guo H Z， Guo D A. J. Chromatogr. B， 2004， 808（2）： 177-183
13 Cui M， Song F， Zhou Y， Liu Z， Liu S. Rapid Commun. Mass Spectrom.， 2000， 14（14）： 1280-1286
14 Zu Y G， Yan M M， Fu Y J， Liu W， Zhang L， Gu C B， Efferth T. J. Sep. Sci.， 2009， 32（4）： 517-525
15 Bompadre S， Tagliabracci A， Battino M， Giorgetti R. J. Chromatogr. B， 2008， 863（1）： 177-180
Simultaneous Quantification of Notoginsenoside R1 and Ginsenoside
Rg1， Re， Rb1 in Tooth Paste by Liquid Chromatography Coupled
with Fully Automated Online Twodimensional Column Switching Method
ZHANG Yan1，2， MA XiaoFei2， L Pin1， CONG Bin*1
1（Hebei Key Laboratory of Forensic Medicine， Department of Forensic Medicine，
Hebei Medical University， Shijiazhuang 050017， China）
2（Hebei Key Laboratory of Food Safety， Hebei Food Inspection and Research Institute， Shijiazhuang 050091， China）
Abstract A novel automated method for simultaneous quantification of notoginsenoside R1 and ginsenoside Rg1， Re， Rb1 in tooth paste was developed using twodimensional columnswitching chromatography with dual gradient liquid chromatography system coupled with UV detector. The determination of notoginsenoside R1 and ginsenoside Rb1 and the purification of ginsenoside Rg1， Re were accomplished in the onedimensional column. The separation of ginsenoside Rg1， Re was finished on the twodimensional column. Syncronis C18 was used as the onedimensional column with the mobile phase of acetonitrile and water， and ODS C18 was employed as the twodimensional column with the mobile phase was as same as in onedimensional column. The detection wavelength was set at 203 nm. The determination was completed in 30 min. The good linearities of notoginsenoside R1 and ginsenoside Rg1， Re， Rb1 were proved respectively in the range of 0.5-200 mg/L and the correlation coefficients （R2） were 0.9994， 0.9996， 0.9995 and 0.9994 respectively. The mean recoveries of notoginsenoside R1 and ginsenoside Rg1， Re， Rb1 were 86.4%-95.1%. It was proved that this method could greatly improve the efficiency of sample analysis.
Keywords High performance liquid chromatography； Twodimensional separation； Notoginsenoside； Ginsenoside
（Received 20 August 2014； accepted 7 October 2014
14 Zu Y G， Yan M M， Fu Y J， Liu W， Zhang L， Gu C B， Efferth T. J. Sep. Sci.， 2009， 32（4）： 517-525
15 Bompadre S， Tagliabracci A， Battino M， Giorgetti R. J. Chromatogr. B， 2008， 863（1）： 177-180
Simultaneous Quantification of Notoginsenoside R1 and Ginsenoside
Rg1， Re， Rb1 in Tooth Paste by Liquid Chromatography Coupled
with Fully Automated Online Twodimensional Column Switching Method
ZHANG Yan1，2， MA XiaoFei2， L Pin1， CONG Bin*1
1（Hebei Key Laboratory of Forensic Medicine， Department of Forensic Medicine，
Hebei Medical University， Shijiazhuang 050017， China）
2（Hebei Key Laboratory of Food Safety， Hebei Food Inspection and Research Institute， Shijiazhuang 050091， China）
Abstract A novel automated method for simultaneous quantification of notoginsenoside R1 and ginsenoside Rg1， Re， Rb1 in tooth paste was developed using twodimensional columnswitching chromatography with dual gradient liquid chromatography system coupled with UV detector. The determination of notoginsenoside R1 and ginsenoside Rb1 and the purification of ginsenoside Rg1， Re were accomplished in the onedimensional column. The separation of ginsenoside Rg1， Re was finished on the twodimensional column. Syncronis C18 was used as the onedimensional column with the mobile phase of acetonitrile and water， and ODS C18 was employed as the twodimensional column with the mobile phase was as same as in onedimensional column. The detection wavelength was set at 203 nm. The determination was completed in 30 min. The good linearities of notoginsenoside R1 and ginsenoside Rg1， Re， Rb1 were proved respectively in the range of 0.5-200 mg/L and the correlation coefficients （R2） were 0.9994， 0.9996， 0.9995 and 0.9994 respectively. The mean recoveries of notoginsenoside R1 and ginsenoside Rg1， Re， Rb1 were 86.4%-95.1%. It was proved that this method could greatly improve the efficiency of sample analysis.
Keywords High performance liquid chromatography； Twodimensional separation； Notoginsenoside； Ginsenoside
（Received 20 August 2014； accepted 7 October 2014
14 Zu Y G， Yan M M， Fu Y J， Liu W， Zhang L， Gu C B， Efferth T. J. Sep. Sci.， 2009， 32（4）： 517-525
15 Bompadre S， Tagliabracci A， Battino M， Giorgetti R. J. Chromatogr. B， 2008， 863（1）： 177-180
Simultaneous Quantification of Notoginsenoside R1 and Ginsenoside
Rg1， Re， Rb1 in Tooth Paste by Liquid Chromatography Coupled
with Fully Automated Online Twodimensional Column Switching Method
ZHANG Yan1，2， MA XiaoFei2， L Pin1， CONG Bin*1
1（Hebei Key Laboratory of Forensic Medicine， Department of Forensic Medicine，
Hebei Medical University， Shijiazhuang 050017， China）
2（Hebei Key Laboratory of Food Safety， Hebei Food Inspection and Research Institute， Shijiazhuang 050091， China）
Abstract A novel automated method for simultaneous quantification of notoginsenoside R1 and ginsenoside Rg1， Re， Rb1 in tooth paste was developed using twodimensional columnswitching chromatography with dual gradient liquid chromatography system coupled with UV detector. The determination of notoginsenoside R1 and ginsenoside Rb1 and the purification of ginsenoside Rg1， Re were accomplished in the onedimensional column. The separation of ginsenoside Rg1， Re was finished on the twodimensional column. Syncronis C18 was used as the onedimensional column with the mobile phase of acetonitrile and water， and ODS C18 was employed as the twodimensional column with the mobile phase was as same as in onedimensional column. The detection wavelength was set at 203 nm. The determination was completed in 30 min. The good linearities of notoginsenoside R1 and ginsenoside Rg1， Re， Rb1 were proved respectively in the range of 0.5-200 mg/L and the correlation coefficients （R2） were 0.9994， 0.9996， 0.9995 and 0.9994 respectively. The mean recoveries of notoginsenoside R1 and ginsenoside Rg1， Re， Rb1 were 86.4%-95.1%. It was proved that this method could greatly improve the efficiency of sample analysis.
Keywords High performance liquid chromatography； Twodimensional separation； Notoginsenoside； Ginsenoside
（Received 20 August 2014； accepted 7 October 2014
