Einsatz der Hochleistungsflüssigkeitschromatographie für die Bestimmung und Isolierung von Capsaicinoiden, Phenolglykosiden und Saponinen

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1 Research Collection Doctoral Thesis Einsatz der Hochleistungsflüssigkeitschromatographie für die Bestimmung und Isolierung von Capsaicinoiden, Phenolglykosiden und Saponinen Author(s): Soldati, Fabio Publication Date: 1979 Permanent Link: Rights / License: In Copyright NonCommercial Use Permitted This page was generated automatically upon download from the ETH Zurich Research Collection. For more information please consult the Terms of use. ETH Library

2 Diss ETH 6347 Einsatz der Hochleistungsflüssigkeitschromatographie für die Bestimmung und Isolierung von Capsaicinoiden, Phenolglykosiden und Saponinen ABHANDLUNG zur Erlangung des Titels eines Doktors der Naturwissenschaften der Eidgenössischen Technischen Hochschule Zürich vorgelegt von FABIO SOLDATI eidg. dipl. Apotheker geboren am 28. Dezember 1948 von Cimadera (TI) Angenommen auf Antrag von Prof. Dr. 0. Sticher, Referent Prof. Dr. X. Perlia, Korreferent 1979

3 ZUSAMMENFASSUNG Im allgemeinen Teil wurde ein kurzer Ueberblick über die Wahl eines Trennsystems in der HPLC gegeben. Die theoretischen und praktischen Grundlagen der Adsorptionschromatographie mit HPLCSäulen wurden in diskutiert. bezug auf die neuesten Literaturangaben Im speziellen Teil sind neue HPLCMethoden beschrieben, die wir für die qualitative Analyse und die quantitative Bestim mung von folgenden Substanzen entwickelt haben: Capsaicinoide: Capsaicin, Dihydrocapsaicin, Homodihydrocapsaicin und Nordihydrocapsaicin in Capsicum frutescens L. Phenolglykoside: Arbutin, Hydrochinon, Hydrochinonmonomethyläther und Methylarbutin in Arctostaphylos Uvaursi (L.) Sprengel, Arctostaphylos alpina (L.) Sprengel, Bergenia crassifolia (L.) Fritsch, Calluna vulgaris (L.) Hüll, Vaccinium Vitisidaea L. Saponine: Glycyrrhizinsäure in Glycyrrhiza glabra L. Ginsenoside Rg,, Rg2' Re unc* R^ ^n Panax ginseng CA. Meyer, Panax quinquefolium L. und pharma zeutischen Spezialitäten Ausserdem werden die Grundlagen für die direkte Isolierung von grossen Mengen der Ginsenoside Rg, Re und Rb,, Rb_ mittels präparativer HPLC beschrieben. Das Massenspektrum des TMSiAethers von Ginsenosid Rg0 wird am Schluss der vorliegenden Arbeit diskutiert.

4 171 SUMMARY In the theoretical part of the work, Separation Systems within the HPLC are reviewed. The theoretical and practical rudiments of the adsorption chromatography with HPLCcolumns are discussed with regard to the new literature. In the experimental part, new HPLC methods, elaborated for the qualitative analysis and the quantitative determination of following substances are described: Capsaicinoides: Capsaicin, dihydrocapsaicin, homodihydrocapsaicin and nordihydrocapsaicin in Capsicum frutescens L. A reversedphase System with a ybondapak C, coluitin using lo methanolwater (53 : 47) as mobile phase was developed. The substances were separated in 40 minutes. The detection limit for capsaicin was 100 ng at a signaltonoice ratio 10 : 1. The relative Standard deviation was 0,54 %. The results obtained by HPLC method are well correlated with those obtained by GCand DCcolorimetric methods. Phenolic glycosides: Arbutin, hydroquinone, hydroquinonemonomethylether and methylarbutin in Arctostaphylos Uvaursi (L.) Sprengel, Arctostaphylos alpina (L.) Sprengel, Bergenia crassifolia (L.) Fritsch, Calluna vulgaris (L.) Hüll, Vaccinium Vitisidaea L. A reversedphase System with a ybondapak C.q column using methanolwater (10 : 90) as mobile phase was developed. The substances were separated in 22 min. The detection limit for arbutin was 25 ng at a signaltonoice ratio of 20 : 1. The relative Standard deviation for arbutin was circa 1,6 %. The

5 172 1,5 results obtained by the highperformance liquid Chromatographie method are well correlated with those obtained by titrimetric and colorimetric method. Loiseleuria procumbens (L.) Desv., Rhododendron ferrugineum (L.), Rhododendron hirsutum (L.), Vaccinium Myrtillus (L.), and Vaccinium ulginosum (L.) do not contain the above mentioned phenolic Compounds. Saponins: Glycyrrhizic aeid in Glycyrrhiza glabra L. The best conditions for the quantitative determination of gly cyrrhizic aeid were elaborated. The reversedphasesystem with a ubondapak Cocolumn and methanolwateracetic aeid (60 : 34 : 6) as eluent turned out to be the best one. The detection limit was 1 yg at a signaltonoice ratio of 20 : 1. The rela tive Standard deviation was circa 1,3 %. Ginsenosides Rg,, Rg5/ Re and Rf in Panax ginseng CA. Meyer, Panax quinquefolium L. and in pharmaceutical drug preparations. A uporasil column using nheptanenbutanolacetonitrilwater (1000 : 446 : 132 : 36) as mobile phase was employed. The gin senosides were separated in 40 min. A reversedphasesystem with a ubondapak Cgcolumn using methanolwater (440 : 560) as eluent was suitable for the Separation of ginsenoside Rg. The reversedphasesystem is adoptable for routine analysis of Ginseng extract. The detection limit for Rg with the spectrophotometer LC 55 (PerkinElmer) was at 207 nm 300 ng at a signaltonoice ratio of 10 : 1. The relative Standard deviation is depending of ginsenosides. For ginsenoside 1 %. from the content Contents of 1 % it was circa Besides, conditions for the Isolation of ginsenosides with preparative HPLC in big quantities are reported. Finally, the mass spectrometry of the ginsenoside as Rg? trimethylsilylether is discussed.