黄连中黄连素的半制备分离

黄连是一种从开花植物中提取的天然药物，被认为对治疗结膜炎和口腔炎有有益作用，是一种胃、肠药和抗腹泻药，黄连素。表明黄连中的一种生物碱，具有黄色、苦味，可能具有抗菌、抗炎作用。该LC应用表明，使用常规HPLC分离从黄连粉末提取物中获得的黄连素，使用半制备HPLC放大分离。制备（提取）：  (1) Weigh 0.5 g of powdered Coptis japonica and place in a centrifuge tube.  (2) Add 30 mL of methanol/10% hydrochloride mixture (100/1) and mix for 15 minutes.   (3) Centrifuge (3,000 rpm, 10mim) and decant the supernatant into a 50 mL measuring flask.   (4) Add 20 mL of methanol/10% HCl mixture (100/1) to the residue and repeat the procedure.   (5) Add methanol/10% HCl mixture (100/1) to the collected supernatant in measuring flask and make up to 50 mL.vApplication Note742025H 2/4Semi-preparative Separation of Berberine in Coptis japonica Semi-preparative Separation of Berberin e in Coptis japonic a Introduction Coptis japonica is a natural medic i ne derived from the f l owering plant and is reputed to have benef i cial effects in the t reatment of conj u nct i vi ti s and stomatitis as well as being a stomachic， i ntestinal remedy and antidiarrhei c， Berber i ne. It i s suggested that one of alkaloids contained in Coptis japonica， which has yellow color and bitter taste， may have antibacterial and anti-inflammatory e f fects. This LC applicat i on shows the separation o f berberine obtained from powder extracts of Coptis japonica usi n g conventional HPLC，the separation is scaled-up using semi-preparat i ve HPLC. Keywords Analytical separation， Sem i -preparative separation，Coptis j aponica， Berberine， Neutraceutical ， Natural medicine JASCO Semi Pre p LC. View product i nformation at www.j as coi nc.c o m 28600 Mary's Court， Easton， MD 21601 USA Conventional HPLC Column： YMC-PACK Pro C18 (4.6 mm IDx250 mmL， 5 um) Eluent： 0.1% TFA in Acetonitrile / Water (30/70) Eluent flow rate： 1.0 mL/min Column temp.： 25°C Wavelength： 220~450 nm， 345 nm Injection volume： 10 pL Standard sample： Powdered Coptis japonica (0.5 g/50 mL in methanol / 10% hydrochloric acid (100/1)) Semi-Preparative HPLC Semi-Preparative HPLC Column： YMC-PACK Pro C18 (20 mm IDx 250 mmL， 5 um) Eluent： 0.1% TFA in Acetonitrile / Water (30/70) Eluent flow rate： 15 mL/min Column temp.： 25°℃ Wavelength： 345 nm Injection volume： 5mL Standard sample： Powdered Coptis japonica (0.5 g/50 mL in methanol / 10% hydrochloric acid (100/1)) Preparation (extraction) (1) Weigh 0.5 g of powdered Coptis j aponica and p l ace in a centr i fuge tu b e. (2) Add 30 mL of methanol/10% hydrochlor i de mixture (100/1) and mix for 15 mi n utes. (3) Centrifuge (3，000 rpm， 10mi m ) and decant th e su p ernatant i n to a 50 mL measur i ng f lask. (4) Add 20 m L of methanol/10% HCl m i xt u re (100/1) to t h e r e sidue and repeat the procedure. (5) Add methanol/10% HCI mi xt u re (100/1) to the col l ected supernatant i n measuring f lask a n d make up t o 50 mL.v JASCO INC. Application Note Fig . 1 Structural formula of Berberine. F i g . 1： Structual Formula of Berberi ne Result Fi g . 2. Chromatogram and contour plot of the extracts from Coptis japonica powder using conventional HPLC separation. Usin g the MD-2018 PDA detector and with spectral comparison， the separation of the target compound Berberine f rom the other compone n ts was optimized and good separation was achieved within 15minutes. Fig. 3. Chroma t ogram of the extract from Coptis japonica powder using semi-preparat i ve HPLC sc a led up from the analytical scale method. To optimize recovery of Berberine the injection vol u me was increased.However， t his caused a problem as the elution power of the extraction sol v ent was stronge r than the mobile phase， the target compound was not retained on column. To resolve this， t he sample was diluted five times i n water and 5 mL of the di l uted sample was i n jected. Berberine was retai n ed as shown in Fig. 3. the separation efficiency increased but with a small sac r ifice of peak shape. Fig . 4. shows the fraction collected using the ChromNAV chromatography data system. The peak fraction and sample rack position for the targe t are hi g hl i ghted in green. Fig. 5. shows demonstrates the sample purity chromatogram of this fraction under the same conditions as in Fig. 2. It is confirmed that Berberine was isolated as single component. Fig. 2： Chromat og ram of t he extract f rom Cop ti s j aponica powder JASCO INC. Lsc o Application Library： http：//www.j ascoinc .com/appl i cations Fig. 3： Semi -preparative c h romatogram of t he extract from Copt i s japonica powder Fi g. 4： Collected f rac ti on o f t he extract f rom Copt i s j ap o n ica po wder (C hromNAV screen) Fig.5： Chromatogram of the collected fraction (10 pL Injected)