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. 2021 Mar 16;26(6):1645.
doi: 10.3390/molecules26061645.

Optimization, Validation and Application of HPLC-PDA Methods for Quantification of Triterpenoids in Vaccinium vitis-idaea L

Gabriele Vilkickyte  1 , Lina Raudone  1   2
Affiliations

Optimization, Validation and Application of HPLC-PDA Methods for Quantification of Triterpenoids in Vaccinium vitis-idaea L

Gabriele Vilkickyte et al. Molecules. .

Abstract

Triterpenoids have regained much attention as promising multi-targeting bioactive agents of natural origin in the treatment of numerous disorders. Due to the high potential for phytopharmaceutical development, accurate qualitative and quantitative analysis of triterpenoids for screening and quality control is required. Vaccinium vitis-idaea L. (lingonberry) raw materials have aroused interest as a rich source of triterpenoids. However, currently, no validated, rapid, and easy-to-perform quantification method is available for the routine control of these compounds in lingonberries. This research aimed at developing and validating HPLC-PDA methods for the determination and screening of triterpenoids in extracts of lingonberry leaves, fruits, and flowers. The developed methods were deemed satisfactory by validation, which revealed acceptable analytical specificity, linearity (r2 > 0.9999), precision (RSD < 2%), trueness (94.70-105.81%), and sensitivity (LOD: 0.08-0.65 µg/mL). The real sample analysis demonstrated established methods applicability for quantification of 13 triterpenoids in lingonberries and emphasized differences between raw materials. Lingonberry fruits were distinguished by the richness of ursolic acid; lingonberry flowers by similar profile to fruits, but low content of neutral triterpenoids; whereas lingonberry leaves by the particularly high level of α-amyrin. Thus, the proposed methods proved to be reliable and applicable for quantification and routine analysis of triterpenoids in lingonberry samples.

Keywords: HPLC; Vaccinium vitis-idaea L.; triterpenoids; validation.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Representative HPLC-PDA chromatograms (λ = 205 nm) of extracts of lingonberry (A) leaves, (B) fruits, (C) flowers, and (D) standard mixture, showing the separation of triterpenoid acids and neutral triterpenoids with chromophores. Peak assignments: 1—maslinic acid, 2—corosolic acid, 3—betulinic acid, 4—oleanolic acid, 5—ursolic acid, 6—betulin, 7—erythrodiol, 8—uvaol.
Figure 2
Figure 2
Representative HPLC-PDA chromatograms (λ = 205 nm) of extracts of lingonberry (A) leaves, (B) fruits, (C) flowers, and (D) standard mixture, showing the separation of phytosterol and neutral triterpenoids, which lack chromophores. Peak assignments: 1—lupeol, 2—β-amyrin, 3—β-sitosterol, 4—α-amyrin, 5—friedelin.
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