Extraction Technologies for Medicinal and Aromatic ... - Capacity4Dev
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Extraction Technologies for Medicinal and Aromatic ... - Capacity4Dev

Extraction Technologies for Medicinal and Aromatic ... - Capacity4Dev Extraction Technologies for Medicinal and Aromatic ... - Capacity4Dev

capacity4dev.ec.europa.eu
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30.10.2014 Views

13 COUNTER-CURRENT CHROMATOGRAPHY Examples of some important natural products that have been separated using HSCCC-CPC in the recent past are summarized herein. 13.5.2.1 Purification of Coenzyme Q 10 from Fermentation Extract: HSCCC versus Silica Gel Column Chromatography HSCCC was applied to the purifi cation of coenzyme Q 10 (CoQ 10 ) for the fi rst time. CoQ 10 was obtained from a fermentation broth extract. A non-aqueous two-phase solvent system composed of heptane–acetonitrile– di chloro methane (12:7:3.5, v/v/v) was selected by analytical HSCCC and used for purifi cation of CoQ 10 from 500 mg crude extract. The separation yielded 130 mg CoQ 10 at an HPLC purity of over 99%. The results showed the advantages of HSCCC over an alternative of silica gel chromatography followed by recrystallization. These advantages regard purity, recovery and yield (Table 3). Table 3: Purification of coenzyme Q 10 from fermentation extract: HSCCC vs. silica gel column chromatography with subsequent crystallization Crude extract CoQ 10 purified by silica gel chromatography HPLC purity, % 89.2 96.0 99.2 Absolute purity, % 29.4 93.3 97.8 Recovery, % * – 74.3 88.0 Yield, % * – 23.4 26.4 CoQ 10 purified by HSCCC * Recovery, amount of CoQ 10 in purifi ed product/amount of CoQ 10 in crude extract. Yield,amount of purifi ed product/amount of crude extract (Journal of Chromatography A 1127 (1-2), 15 September 2006, 92-96) 13.5.2.2 Preparative Separation of Gambogic Acid and its C-2 Epimer by HPCCC For the preparative separation of epimers, gambogic acid and epigambogic acid, from Garcinia hanburyi, a two-phase solvent system composed of n-hexane–methanol–water (5:4:1, v/v/v) was used. From 50 mg mixture, 28.2 mg gambogic acid and 18.4 mg epigambogic acid were separated. The purities of both were above 97% as determined by HPLC. The chemical structures were then identified by their 1 H NMR and 13 C NMR spectra. 13.5.2.3 Separation and Purification of 10-deacetylbaccatin III by HSCCC At present, the most promising approach is the semisynthesis of paclitaxel or its analogs from 10-deacetylbaccatin III, a compound available in a relatively high quantity from the foliage of several yew species. HSCCC was used for the separation and purification of 10-deacetylbaccatin III (Figure 11). A crude needle extract (500 mg/5 ml) from Chinese yew (Taxus chinensis) was 228

EXTRACTION TECHNOLOGIES FOR MEDICINAL AND AROMATIC PLANTS first separated with a two-phase solvent system composed of n-hexane–ethyl acetate–ethanol–water (2:5:2:5, v/v). The partially purified fraction was again purified with a different solvent system composed of n-hexane–chloroform– methanol–water (5:25:34:20, v/v). HPLC analysis of the final fraction showed that the purity of 10-deacetylbaccatin (20 mg) was over 98%. Figure 11: Separation and purifi cation of 10-deacetylbaccatin III (DAB) by HSCCC 229

EXTRACTION TECHNOLOGIES FOR MEDICINAL AND AROMATIC PLANTS<br />

first separated with a two-phase solvent system composed of n-hexane–ethyl<br />

acetate–ethanol–water (2:5:2:5, v/v). The partially purified fraction was again<br />

purified with a different solvent system composed of n-hexane–chloro<strong>for</strong>m–<br />

methanol–water (5:25:34:20, v/v). HPLC analysis of the final fraction showed<br />

that the purity of 10-deacetylbaccatin (20 mg) was over 98%.<br />

Figure 11: Separation <strong>and</strong> purifi cation of 10-deacetylbaccatin III (DAB) by HSCCC<br />

229

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