高速逆流色谱原理以及应用

1、关于高速逆流色谱原理及应用第一张，PPT共四十页，创作于2022年6月多级萃取技术第二张，PPT共四十页，创作于2022年6月第三张，PPT共四十页，创作于2022年6月离心分配色谱平面图第四张，PPT共四十页，创作于2022年6月离心分配色谱立体图第五张，PPT共四十页，创作于2022年6月同步行星式逆流色谱第六张，PPT共四十页，创作于2022年6月第七张，PPT共四十页，创作于2022年6月高速逆流色谱运行平面图第八张，PPT共四十页，创作于2022年6月第九张，PPT共四十页，创作于2022年6月分离区分离区混合区第十张，PPT共四十页，创作于2022年6月第十一张，PPT共四十页，创

2、作于2022年6月HSCCC 系统连接工作图第十二张，PPT共四十页，创作于2022年6月1、HPLC方法的建立尽可能使各峰得到良好的分离2、溶剂系统的选择根据样品的溶解特性选择溶剂体系测定分配系数，依据分配系数确定溶剂系统K2/K1:0.5-2.53、HSCCC分离条件优化色谱柱转速；固定相的保留；流动相流速；分离温度。4、HPLC测定收集组分的纯度HSCCC方法的建立过程:第十三张，PPT共四十页，创作于2022年6月丁醇-醋酸-水丁醇-甲醇-水(缓冲液)丁醇-乙酸乙酯-水(缓冲液)乙酸乙酯-水(缓冲液)氯仿-甲醇-水(毒)甲基异丁基酮-丙酮-水正己烷-乙酸乙酯-甲醇-水(石油醚-乙酸乙酯

3、-甲醇-水)正己烷-乙腈正己烷-甲醇-水正己烷-乙酸乙酯-甲醇-水(石油醚-乙酸乙酯-甲醇-水)水溶性和强极性中等极性(氯仿)弱极性(正己烷)HSCCC常用溶剂系统（依据组分的极性和溶解性）第十四张，PPT共四十页，创作于2022年6月Preparative Isolation of Six Major Saponins from Platycodi Radix by High-speed Counter-current ChromatographyYoung Wan Ha and Yeong Shik KimABSTRACT:Introduction Platycosides, the pr

4、imary constituents of Platycodi Radix, are known to have numerous and varied biological activities, exerting anti-inflammation, anti-allergy, anti-tumour, anti-obesity and anti-hyperlipidemia effects. However, effective methods for isolating and purifying platycosides from Platycodi Radix are not cu

5、rrently available.Objective To develop an efficient method for the preparative separation of six platycosides from Platycodi Radix by high speed counter-current chromatography (HSCCC) coupled with an evaporative light scattering detection (ELSD) system.Methodology Preparative separation was performe

6、d by water extraction using reversed-phase C18 column chromatography on an HSCCC-ELSD system. A two-phase solvent system comprised hexanen-butanolwater (1:40:20, v/v) and (1:10:5, v/v) was employed. Two other key parameters, revolution speed of the separation column and flow-rate of the mobile phase

7、, were also investigated for optimum HSCCC performance. Each peak fraction obtained from separation of the platycosides was collected according to the ELSD elution profile and determined by HPLC.Results Using the described method, six platycosides, all with purities of over 94%, could be isolated fr

8、om 300mg of the platycoside-enriched fraction. Their structures were characterized by electrospray ionisation mass spectrometry (ESI-MS), 1HNMR and 13C-NMR.第十五张，PPT共四十页，创作于2022年6月IntroductionExperimentalApparatus.Reagents and materials.Preparation of crude sample.Measurement of partition coefficient

9、sPreparation of the two-phase solvent system and sample solution.HSCCC separationSelection of the two-phase solvent system and HSCCC conditionsResults and DiscussionHPLC analysis and identification of HSCCC peak fractionsStructural identificationAcknowledgementsReferences第十六张，PPT共四十页，创作于2022年6月第十七张，

10、PPT共四十页，创作于2022年6月第十八张，PPT共四十页，创作于2022年6月高速逆流色谱的延伸应用第十九张，PPT共四十页，创作于2022年6月实例1 pH区带CCC分离同分异构体HSCCC条件：溶剂系统:正丁醇-水(1:1)固 定 相:上相pH=0.5(HCl)流 动 相:下相pH=10.7(NH3)流 速:2.0 ml min-1转 速:715 rpm检测波长:206 nm样 品 量:230 mg 粗提物机 型:pH-zone-refining CCC固定相保留： 60%1: 3-sulfophthalic acid 99.02: 4-sulfophthalic acid 99. 0

11、Journal of Chromatography A, 966 (2002) 111118pH区带逆流色谱分离中药单体中应用第二十张，PPT共四十页，创作于2022年6月实例2 CCC分离蛇床子HSCCC条件：溶剂系统:正己烷-乙酸乙酯-乙醇-水(5:5 :5:5)流 速: 0 180 min, 1ml min1 180480 min, 2ml min1转 速:1000 rpm检测波长:254 nm样 品 量:500 mg 粗提物机 型:CCC-1000相 保 留:58%A: bergapten 96.5% 45.8 mg B: imperatorin 98.2% 118.3 mg 92.1

12、 、93.7% Journal of Chromatography A, 1061 (2004) 5154逆流色谱分离中药应用(V流动相)第二十一张，PPT共四十页，创作于2022年6月实例3 CCC分离短瓣金莲花黄酮苷HSCCC条件：溶剂系统:乙酸乙酯-正丁醇-水(2:1:3)流 速: 0 190 min, 1.5ml min1 190480 min, 2.5ml min1转 速:800 rpm检测波长:254 nm样 品 量:500 mg 粗提物机 型:CCC-1000保 留 值:49%1: quercetin-3-O-neohesperidoside 85.1% 9.3 mg2: ori

13、entin(荭草素) 97% 95.8 mg 3: 未知化合物 97% 4: vitexin(牡荆素) 97% 11.6 mg Journal of Chromatography A, 1092 (2005) 216221逆流色谱分离中药应用(V流动相)1.5 ml/min2.5 ml/min第二十二张，PPT共四十页，创作于2022年6月贯叶连翘中金丝桃苷HSCCC-HPLC-DAD联用分离中药应用第二十三张，PPT共四十页，创作于2022年6月三相CCC分离15种化合物逆流色谱分离中药应用(三相溶剂系统)HEXMeOAcACN-Water第二十四张，PPT共四十页，创作于2022年6月实例

14、6 二维CCC分离冬凌草HSCCC条件：溶剂系统:一维:正己烷-乙酸乙酯-甲醇-水 1:5:1:5二维:正己烷-乙酸乙酯-甲醇-水 3:5:3:5流 速 :2ml min1 转 速 :800 rpm样 品 量 :250 mg 粗提物机 型: 2D-CCC保 留 值:一维: 40% 二维: 63% 1: oridonin (甲素) 95% 2: ponicidin (乙素) 95%Journal of Chromatography A, 1146 (2007) 125130二维逆流色谱分离中药应用第二十五张，PPT共四十页，创作于2022年6月实例7 分离白花前胡中香豆素HSCCC条件：溶剂系统

15、:石油醚-乙酸乙酯-甲醇-水 5:5:6:4流 速 :2ml min1 转 速 :900 rpm样 品 量 :250 mg 粗提物机 型 :TBE-300保 留 值 :54.3% 1: 7.0 mg 99.3% 2: 7.4 mg 96.1% 3: 12.5mg 96.5% 4: 42.1mg 99.3% 5: 9.8 mg 99.7% 6: 65.2 mg 98.1% 7: 31.7mg, 92.8%Journal of Chromatography B, 877 (2009) 25712578HSCCC-ESI-MSn分离中药应用第二十六张，PPT共四十页，创作于2022年6月扩展和想象湖

16、州:活猪走进去 鲜肉运出来 全机械化操作 中国猪业网2010-12-27 由于该定点屠宰迁建项目采用自动化同步流水线加工，活猪进去，出来便是新鲜的猪肉，减少了手工的环节和过程，猪肉更卫生，肉品质量也会有大幅提高。 第二十七张，PPT共四十页，创作于2022年6月Application of accelerated solvent extraction coupled with high-performance counter-current chromatography to extraction and online isolation of chemical constituents fr

17、om Hypericum perforatum L.Yuchi Zhang, Chunming Liu, Min Yu, Zhengkun Zhang, Yanjuan Qi, Jing Wang, Guimei Wu, Sainan Li, Jing Yu, Yang HuJournal of Chromatography A, 1218 (2011) 28272834第二十八张，PPT共四十页，创作于2022年6月Hypericum perforatum L., a typical traditional Chinese medicine, is used all over the wor

18、ld for the treatment of depression. Additionally, it has been reported that the extracts of H. perforatum may have an inhibitory effect, on human bladder cancer cells , and act as an antioxidant and anticonvulsant. The extracts of H. perforatum contain many constituents with documented biological ac

19、tivity such as quinic acids, phloroglucinols and a broad range of flavonoids . Exposure of the extracts of H. perforatum to light may lead to the degradation of phloroglucinols, which are extremely sensitive to oxidation and unstable in solution on exposure to air, therefore phloroglucinols are diff

20、icult to separate and isolate by conventional method. In this case, amethod combining an extraction system with an isolation system online to avoid the exposure of the extracts to air and light is urgently needed.Introduction第二十九张，PPT共四十页，创作于2022年6月The diagram presents two setups (B) enabling hyphen

21、ation of ASE and HPCCC第三十张，PPT共四十页，创作于2022年6月电视剧东北一家人背景：汽车厂家属区开辟一片荒地，打算承包给个人，由个人决定所要开发的娱乐项目。甲（牛小伟，吕小品扮演）：打算建设过山车。乙（牛继红，金珠扮演） ：打算开发滑梯和秋千相关的项目。丙（孙明，巩汉林扮演） ：认为甲和乙所开发的项目太过时，小儿科，丙的思路是在 过山车上打滑梯，荡秋千。扩展和想象第三十一张，PPT共四十页，创作于2022年6月第三十二张，PPT共四十页，创作于2022年6月第三十三张，PPT共四十页，创作于2022年6月Completed preparative separatio

22、n of alkaloids from Cephaltaxus fortunine by step-pH-gradient high-speed counter-current chromatographyZhonghua Liu, Qizhen Du, Kuiwu Wang, Lili Xiu, Guanglei SongJournal of Chromatography A, 1216 (2009) 46634667第三十四张，PPT共四十页，创作于2022年6月Chemical structure of cephalotaxine-type alkaloids第三十五张，PPT共四十页，

23、创作于2022年6月The HSCCC experimentwas performed with a procedure: The column was first entirely filled with the organic stationary phase containing TFA. This was followed by sample injection. Then the aqueous phase containing 2% NH4OH (eluter base) was pumped into the inlet of the column at a flow-rate

24、of 3.0 mL/min in the head to tail elution mode, while the apparatus was rotated at 750 rpm.After peaks I and II were eluted out of the column, the mobile phase was changed to the aqueous phase containing 0.2% NH4OH until peak III were eluted out. Following this, the mobile phase was changed to the aqueous phase containing 0.01% TFA to elute peaks VI and VII.HSCCC separation第三十六张，PPT