人参中人参皂苷的半制备分离

人参是一种天然药材，来源于大麻油科多年生草本植物，俗称亚洲人参或朝鲜人参。据广泛报道，人参具有许多益处，包括疲劳恢复、退热、血压控制（低血压和高血压）、抗炎/抗菌作用（胃和十二指肠溃疡）、止血、强心作用、抗肿瘤作用（抗癌作用）、糖尿病护理（血糖水平控制和胰岛素促分泌）。人参含有大量的人参苷，这是一种皂苷。人参皂甙Rb1具有中枢抑制作用，人参皂甙Rg1具有中枢兴奋作用，其抗疲劳和镇静作用已有报道。该HPLC应用描述了在常规反相HPLC中使用梯度洗脱分离人参皂苷Rb1和Rg1的方法的开发，该方法可以放大为半制备HPLC。Application Note 2/4Application Note Semi-preparative Separation of Ginsenoside in Ginseng Semi-preparativeSeparation o f Ginsenoside in Ginseng Gi n seng i s a natural medicine derived f rom the araliaceae herbaceous perennial more commonly known as Asia n ginseng or Korean ginseng . Ginseng has been widely reported a s having many benefits includi n g recovery from fatigue， pyreto l y s i s ， blood pressure control (low-and high-blood pressure)， anti-inflammatory /antibacterial action (gast r ic and duodenal ulcer)， hemostasis， cardiotonic action， anti -tu m or action (anti-cancer action)， diabetes care (blood-sugar level control and insulin secretagogue). Ginseng contains a large amount of ginsenoisides -a type of saponin.Gi n senoiside Rb1 has central depressant action and ginsenoiside Rg1 has central excitatory action，and their anti-fatig u e and sedative action have been reported. This HPLC application descr i bes the development of a separat i on method for ginsenoside Rb1 and Rg1 using gradient elution i n conventional reverse phase HPLC which can be scaled-up to semi-prepa r at i ve HPLC. Analytical separation， Semi -preparative separation，Copt i s japonica， Berberine， Neutraceut i cal ， Natural medicine JASCO Semi Pre p LC. View product i nformation at www.j a s coi nc.c o m Experimenta l Equipment Conventional HPLC Eluent Pump： PU-2089 Autosampler： AS-2057 Column oven： CO-2060 Detector： MD-2018 min(50/50)-> min(80/20) Semi-Preparative HPLC Eluent Pump： PU-2086 (x2) Mixer： MX-2080-32 (With 10 mL chamber) Autosampler： AS-2058 Column oven： CO-2060 Detector： MD-2018 min(50/50)-> min(80/20) Chromatography datasystem： Fraction collector： ADVANTEC SCF 122SC Fraction collector FC-2088-30 controller： Conditions Conventional HPLC Column： YMC-PACK Pro C18 (4.6 mm IDx250 mmL， 5 pm) Eluent： A； Water， B； Acetonitrile， linear gradient Gradient (A/B)， 0 min(80/20)->15 condition： 20 min (50/50) ->20.1 1 cycle； 40 min Eluent flow rate： 1.0 mL/min Column temp.： 25°℃ Wavelength： 200~4 50 nm， 203 nm Injection volume： 20 pL Standard sample： Powdered Ginseng (1.0g/50mL in 60% methanol) Semi-Preparative HPLC Column： YMC-PACK Pro C18 (20 mm IDx250 mmL， 5 pm) Eluent： A； Water， B； Acetonitrile， linear gradient Gradient (A/B)， 0 min(80/20)->15 condition： Eluent flow rate： 15 mL/min Column temp.： 25°℃ Wavelength： 203nm Injection volume： 5mL Standard sample： Powdered Ginseng (1.0g/50mL in 60% methanol) (1) Weigh 1.0 g of powdered ginseng and place i n a centrifuge tube. (2) Add 30 m L of 60% methanol a n d mi x for 15 m i n utes. (3) Cent r ifuge (3，000 rpm， 10mim) and decant the supernatant i nto a 50 mL measu r i ng f lask (4) Add 20 mL of 60% methanol to th e residue a n d repea t the procedure. (5) Ad d 60% methanol to collected supernatant i n m easuri n g f l ask and make up to 50 mL Application Note Fig. 1 Structural formula of Ginsenosides. Fig . 1： Ginsenoside Rb 1 Fig . 1： G i nse n oside Rg1 Result Fig. 2 Chromatogram and contour plot of extracts from ginseng powder separated using conventional HPLC. Since the retention of ginsenoiside Rg1 and Rb1 are different ， the separation condi t ions have been determined using gradien t el u tion. The MD-2018 PDA detector and spectral comparison was used to optimize the separation of the target compounds from other components； ginsenois i des Rg1 and Rb1 were clear l y separated within 16 minutes. Fi g. 2： Chromato g ram o f the ex t ract f r om Ginseng powder Fig . 3. shows the chromatogram o f the ginseng powder separated using a semi-preparative HPLC scaled-up from analytical. In order to maximize the recovery of t he separated ginsenoisides a 5 mL sample volu m e was injected. F i g. 4. shows the f raction display in the ChromNAV chromatography data system. The peaks fractions and sample rack position for the target com p ounds are hig h lighted i n green. Fig. 5. shows chromatograms of a pur i ty check of the recovered f r a ctions analyzed using t he same conditions as i n Fig . 2. Ginsenoiside Rb1 and a minor component were n ot completely resolved i n the sem i -prep separation and small contaminant peak can be observed just after the main peak， but it was confirmed that each compound was clearly isolated. Fig .3： Semi -preparative chrom a togram of an extract from Ginseng powder Fig . 4： Peak f ract i on result o f an extrac t from Ginse n g P owder (C hromNAV Screen) Fig.5： Chromatogram of the collected fraction (10 pL Injected) Applicatio n Library： http：//www.j ascoinc .com/appl i cations