張巖等

摘 要 使用雙梯度液相色譜系統(tǒng)紫外檢測(cè)器，建立了二維液相色譜法全自動(dòng)快速同時(shí)測(cè)定牙膏中三七皂苷R1、人參皂苷Rg1、Re和Rb1的含量。樣品經(jīng)超聲提取后，以Syncronis C18為一維分析柱，ODS C18為二維分析柱，利用一維色譜柱完成三七皂苷R1和人參皂苷Rb1分離測(cè)定以及人參皂苷Rg1和人參皂苷Re的凈化； 利用二維色譜柱完成人參皂苷Rg1和人參皂苷Re的分析。一維分析和二維分析均以乙腈水體系作為流動(dòng)相，梯度洗脫，檢測(cè)波長(zhǎng)為203 nm，整個(gè)分析過(guò)程僅需30 min。三七皂苷R1、人參皂苷Rg1、Re和Rb1在0.5～200 mg/L范圍內(nèi)線性良好，相關(guān)系數(shù)R2分別為0.9994， 0.9996， 0.9995和0.9994，平均回收率均在86.4%～95.1%之間。本方法簡(jiǎn)便快速，測(cè)定結(jié)果準(zhǔn)確可靠，可用于牙膏中三七皂苷R1、人參皂苷Rg1、Re和Rb1含量的測(cè)定。

3.3 一維流動(dòng)相的選擇

考察了流動(dòng)相為乙腈水體系和甲醇水體系對(duì)一維

色譜保留時(shí)間及峰形的影響，結(jié)果表明，使用甲醇水為流動(dòng)相三七皂苷R1、人參皂苷Rg1和Re三者不能達(dá)到基線分離，且峰形較寬（圖4）。無(wú)法確定人參皂苷Rg1和Re切入二維色譜柱的時(shí)間，因色譜峰形過(guò)寬，需要切入時(shí)間長(zhǎng)，造成流動(dòng)相比例不匹配，容易引起壓力波動(dòng)，從而影響到二維色譜中目標(biāo)物保留時(shí)間的漂移。采用乙腈水作為一維色譜流動(dòng)相時(shí)，雖然人參皂苷Rg1和Re重合在一起，但二者能夠與三七皂苷R1達(dá)到基線分離分析，且峰形良好，峰窄，從而確定了二維分析的切入時(shí)間，故選擇了乙腈水體系作為一維流動(dòng)相（圖4）。通過(guò)優(yōu)化實(shí)驗(yàn)條件發(fā)現(xiàn)乙腈水體系應(yīng)用于二維色譜能夠?qū)⑷藚⒃碥誖g1和Re分離，為了使流動(dòng)相匹配，避免壓力波動(dòng)，所以二維流動(dòng)相選擇乙腈水體系。

在一維分析中三七皂苷R1和人參皂苷Rb1較容易分離，所以在一維色譜中采用高比例的有機(jī)相和細(xì)粒徑的色譜柱，提高分析速度，使在一維色譜中較難分離且存在基質(zhì)干擾的人參皂苷Rg1和Re切換到二維色譜柱中繼續(xù)分離，常規(guī)色譜分析4種物質(zhì)需60 min以上，本方法僅需30 min，提高了分析速度，同時(shí)解決了人參皂苷Rg1和Re存在干擾和分離的難題。

實(shí)驗(yàn)結(jié)果表明， 通過(guò)在線二維柱切換，雙梯度洗脫，簡(jiǎn)化了分析過(guò)程，提高了分析速度，有效去除了復(fù)雜基質(zhì)的干擾。本方法可作為牙膏中三七皂苷R1、人參皂苷Rg1， Re和Rb1的常規(guī)檢測(cè)方法，為監(jiān)管部門規(guī)范管理口腔衛(wèi)生產(chǎn)品市場(chǎng)提供了技術(shù)支撐。

References

1 WANG Qiang， JIANG YingQiao， MA ShiPing， DANG XueDong. China Journal of Chinese Materia Medica， 2000， 25（10）： 617-618

王 強(qiáng)， 江英橋， 馬世平， 黨學(xué)東. 中國(guó)中藥雜志， 2000， 25（10）： 617-618

2 ZHANG XiaoPing， XU ShiJie. Chin. JMAP， 2006， 23（3）： 219-220

張小平， 徐世杰. 中國(guó)現(xiàn)代應(yīng)用藥學(xué)雜志， 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， 郭曉煥. 中國(guó)醫(yī)藥科學(xué)， 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

沙東旭， 張滿來(lái). 中國(guó)藥雜志， 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