高速逆流色譜原理以及應(yīng)用

1、關(guān)于高速逆流色譜原理及應(yīng)用第一張，PPT共四十頁，創(chuàng)作于2022年6月多級萃取技術(shù)第二張，PPT共四十頁，創(chuàng)作于2022年6月第三張，PPT共四十頁，創(chuàng)作于2022年6月離心分配色譜平面圖第四張，PPT共四十頁，創(chuàng)作于2022年6月離心分配色譜立體圖第五張，PPT共四十頁，創(chuàng)作于2022年6月同步行星式逆流色譜第六張，PPT共四十頁，創(chuàng)作于2022年6月第七張，PPT共四十頁，創(chuàng)作于2022年6月高速逆流色譜運行平面圖第八張，PPT共四十頁，創(chuàng)作于2022年6月第九張，PPT共四十頁，創(chuàng)作于2022年6月分離區(qū)分離區(qū)混合區(qū)第十張，PPT共四十頁，創(chuàng)作于2022年6月第十一張，PPT共四十頁，創(chuàng)

2、作于2022年6月HSCCC 系統(tǒng)連接工作圖第十二張，PPT共四十頁，創(chuàng)作于2022年6月1、HPLC方法的建立盡可能使各峰得到良好的分離2、溶劑系統(tǒng)的選擇根據(jù)樣品的溶解特性選擇溶劑體系測定分配系數(shù)，依據(jù)分配系數(shù)確定溶劑系統(tǒng)K2/K1:0.5-2.53、HSCCC分離條件優(yōu)化色譜柱轉(zhuǎn)速；固定相的保留；流動相流速；分離溫度。4、HPLC測定收集組分的純度HSCCC方法的建立過程:第十三張，PPT共四十頁，創(chuàng)作于2022年6月丁醇-醋酸-水丁醇-甲醇-水(緩沖液)丁醇-乙酸乙酯-水(緩沖液)乙酸乙酯-水(緩沖液)氯仿-甲醇-水(毒)甲基異丁基酮-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)水溶性和強(qiáng)極性中等極性(氯仿)弱極性(正己烷)HSCCC常用溶劑系統(tǒng)（依據(jù)組分的極性和溶解性）第十四張，PPT共四十頁，創(chuàng)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.第十五張，PPT共四十頁，創(chuàng)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第十六張，PPT共四十頁，創(chuàng)作于2022年6月第十七張，

10、PPT共四十頁，創(chuàng)作于2022年6月第十八張，PPT共四十頁，創(chuàng)作于2022年6月高速逆流色譜的延伸應(yīng)用第十九張，PPT共四十頁，創(chuàng)作于2022年6月實例1 pH區(qū)帶CCC分離同分異構(gòu)體HSCCC條件：溶劑系統(tǒng):正丁醇-水(1:1)固 定 相:上相pH=0.5(HCl)流 動 相:下相pH=10.7(NH3)流 速:2.0 ml min-1轉(zhuǎn) 速:715 rpm檢測波長:206 nm樣 品 量:230 mg 粗提物機(jī) 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區(qū)帶逆流色譜分離中藥單體中應(yīng)用第二十張，PPT共四十頁，創(chuàng)作于2022年6月實例2 CCC分離蛇床子HSCCC條件：溶劑系統(tǒng):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轉(zhuǎn) 速:1000 rpm檢測波長:254 nm樣 品 量:500 mg 粗提物機(jī) 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逆流色譜分離中藥應(yīng)用(V流動相)第二十一張，PPT共四十頁，創(chuàng)作于2022年6月實例3 CCC分離短瓣金蓮花黃酮苷HSCCC條件：溶劑系統(tǒng):乙酸乙酯-正丁醇-水(2:1:3)流 速: 0 190 min, 1.5ml min1 190480 min, 2.5ml min1轉(zhuǎn) 速:800 rpm檢測波長:254 nm樣 品 量:500 mg 粗提物機(jī) 型:CCC-1000保 留 值:49%1: quercetin-3-O-neohesperidoside 85.1% 9.3 mg2: ori

13、entin(葒草素) 97% 95.8 mg 3: 未知化合物 97% 4: vitexin(牡荊素) 97% 11.6 mg Journal of Chromatography A, 1092 (2005) 216221逆流色譜分離中藥應(yīng)用(V流動相)1.5 ml/min2.5 ml/min第二十二張，PPT共四十頁，創(chuàng)作于2022年6月貫葉連翹中金絲桃苷HSCCC-HPLC-DAD聯(lián)用分離中藥應(yīng)用第二十三張，PPT共四十頁，創(chuàng)作于2022年6月三相CCC分離15種化合物逆流色譜分離中藥應(yīng)用(三相溶劑系統(tǒng))HEXMeOAcACN-Water第二十四張，PPT共四十頁，創(chuàng)作于2022年6月實例

14、6 二維CCC分離冬凌草HSCCC條件：溶劑系統(tǒng):一維:正己烷-乙酸乙酯-甲醇-水 1:5:1:5二維:正己烷-乙酸乙酯-甲醇-水 3:5:3:5流 速 :2ml min1 轉(zhuǎn) 速 :800 rpm樣 品 量 :250 mg 粗提物機(jī) 型: 2D-CCC保 留 值:一維: 40% 二維: 63% 1: oridonin (甲素) 95% 2: ponicidin (乙素) 95%Journal of Chromatography A, 1146 (2007) 125130二維逆流色譜分離中藥應(yīng)用第二十五張，PPT共四十頁，創(chuàng)作于2022年6月實例7 分離白花前胡中香豆素HSCCC條件：溶劑系統(tǒng)

15、:石油醚-乙酸乙酯-甲醇-水 5:5:6:4流 速 :2ml min1 轉(zhuǎn) 速 :900 rpm樣 品 量 :250 mg 粗提物機(jī) 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分離中藥應(yīng)用第二十六張，PPT共四十頁，創(chuàng)作于2022年6月擴(kuò)展和想象湖

16、州:活豬走進(jìn)去 鮮肉運出來 全機(jī)械化操作 中國豬業(yè)網(wǎng)2010-12-27 由于該定點屠宰遷建項目采用自動化同步流水線加工，活豬進(jìn)去，出來便是新鮮的豬肉，減少了手工的環(huán)節(jié)和過程，豬肉更衛(wèi)生，肉品質(zhì)量也會有大幅提高。 第二十七張，PPT共四十頁，創(chuàng)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第二十八張，PPT共四十頁，創(chuàng)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第二十九張，PPT共四十頁，創(chuàng)作于2022年6月The diagram presents two setups (B) enabling hyphen

21、ation of ASE and HPCCC第三十張，PPT共四十頁，創(chuàng)作于2022年6月電視劇東北一家人背景：汽車廠家屬區(qū)開辟一片荒地，打算承包給個人，由個人決定所要開發(fā)的娛樂項目。甲（牛小偉，呂小品扮演）：打算建設(shè)過山車。乙（牛繼紅，金珠扮演） ：打算開發(fā)滑梯和秋千相關(guān)的項目。丙（孫明，鞏漢林扮演） ：認(rèn)為甲和乙所開發(fā)的項目太過時，小兒科，丙的思路是在 過山車上打滑梯，蕩秋千。擴(kuò)展和想象第三十一張，PPT共四十頁，創(chuàng)作于2022年6月第三十二張，PPT共四十頁，創(chuàng)作于2022年6月第三十三張，PPT共四十頁，創(chuàng)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第三十四張，PPT共四十頁，創(chuàng)作于2022年6月Chemical structure of cephalotaxine-type alkaloids第三十五張，PPT共四十頁，

23、創(chuàng)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第三十六張，PPT