Determination of triterpenic acids in fruits by a novel high performance liquid chromatography method with high sensitivity and specificity

Application of Response Surface Methodology for Optimisation of Simultaneous UHPLC-PDA Determination of Oleanolic and Ursolic Acids and Standardisation of Ericaceae Medicinal Plants

“…In comparison with the previously developed HPLC methods with UV- (Table 5) and ELS-detection [25], as well as with the only one available to date UHPLC method [28] (Table 5), the sensitivity was highly improved. Lower LODs and LOQs could have been obtained for native OA and UA (43-95 ng/mL; 0.017-0.038 ng) with MS-detection [26], and for fluorophore derivatives (0.8-6.3 ng/mL; 0.004-0.063 ng) with FL-detection [17,[22][23][24]. Stability of the standards (expressed as recovery of the initial concentration) was tested at 4 • C during 30 days in five-day intervals.…”

“…Considering the earlier studies, C18 is the stationary phase that is most commonly used in the LC separation of OA and UA [8,[15][16][17][18][19][20][21]26,38]. This is hardly surprising, as RP-18 columns are also the most popular ones when it comes to plant material standardisation in general and, therefore, easily available in laboratories performing such studies.…”

“…Although some significant effects of organic solvents and pH of the mobile phase on selectivity have been reported [16], the stationary phase efficiency is believed to be the most important factor determining the resolution of the overlapping bands of native acids. Subsequently, even for relatively simple plant matrices, the use of long chromatographic columns and analysis time of over 15 min, and sometimes over 30 min, are usually required for satisfactory separation [15][16][17][18][19]. As most of the methods utilize HPLC equipment with ultraviolet (UV) detectors, reduced sensitivity may be also an issue, as triterpenes, in general, exhibit low absorption of ultraviolet light.…”

“…To address these problems various methods have been applied. Adding of separation facilitating substances, like cyclodextrins [20,21], might improve the resolution, as might pre-column derivation [17,[22][23][24], which is sometimes used as a solution for low sensitivity. Better limits of detection may be also achieved with detectors other than UV, e.g., evaporative light scattering (ELS) or mass selective (MS) detectors [23][24][25][26].…”

Analysis of triterpenoids and phytosterols in vegetables by thin-layer chromatography coupled to tandem mass spectrometry

“…Presence of isomeric triterpenoids in plant cuticular waxes renders the determination of triterpenoids even more difficult. Among the separation techniques, thin-layer (TLC) [13][14][15][16], supercritical fluid (SFC) [17], gas (GC) [18][19][20][21][22][23][24] and highperformance liquid (HPLC) [13,14,[23][24][25][26][27][28][29][30][31][32][33] chromatography and capillary electrophoresis (CE) [34] have been used in their analysis. GC is favorable for the separation of positional triterpenoid isomers, and coupled to flame ionizaton (FID) [18,19,[21][22][23][24] and mass spectrometric (MS) detection [18][19][20]22,24] has been widely used for their qualitative [18][19][20]22,24] and quantitative analysis [18,19,[21][22]…”