Preparative TLC Fractionation of Triterpenoids

Evaporated diethyl ether extracts were fractionated by adsorption preparative TLC on 20 cm × 20 cm glass plates coated manually with silica gel 60H (Merck, Darmstad, Germany). The solvent system, chloroform:methanol 97:3 (v/v), was applied for developing. The fractions of triterpenoid acids and free (non-esterified) steroids and neutral triterpenoids were localized on the plates by comparison to the appropriate standards, as described earlier [16 (link),24 (link)]. Fractions were eluted from the gel in diethyl ether. Subsequently, fractions containing free neutral triterpenes and sterols (R_F 0.3–0.9) were directly analyzed by GC-MS, whereas the fractions containing triterpene acids (R_F 0.2–0.3) were methylated with diazomethane. The average recovery of α-amyrin, uvaol, stigmasterol, and ursolic acid methyl ester from preparative TLC plates was 98.6, 97.2, 98.9, and 96.1%, respectively [25 (link)].

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Dashbaldan S., Rogowska A., Pączkowski C, & Szakiel A. (2021). Distribution of Triterpenoids and Steroids in Developing Rugosa Rose (Rosa rugosa Thunb.) Accessory Fruit. Molecules, 26(17), 5158.

Publication 2021

silica gel 60H

Acids Adsorption Chloroform Diazomethane Diethyl ether Ester Gc ms Methanol Silica gel Solvent Steroids Sterols Stigmasterol Triterpene Ursolic acid Uvaol

Corresponding Organization : University of Warsaw

Other organizations : Mongolian University of Science and Technology

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Variable analysis

independent variables

Solvent system (chloroform:methanol 97:3 (v/v))
Preparative TLC on silica gel 60H plates

dependent variables

Fractionation of evaporated diethyl ether extracts
Elution of fractions containing free neutral triterpenes and sterols (RF 0.3–0.9)
Methylation of fractions containing triterpene acids (RF 0.2–0.3)
GC-MS analysis of fractions containing free neutral triterpenes and sterols

control variables

Size of preparative TLC plates (20 cm × 20 cm)
Manually coated silica gel 60H plates (Merck, Darmstad, Germany)
Comparison to appropriate standards for localization of fractions

controls

Appropriate standards for localization of fractions
Average recovery percentages for α-amyrin, uvaol, stigmasterol, and ursolic acid methyl ester from preparative TLC plates

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