Abstract
An HPLC-PDA method was developed for the determination of the flavonoids in the flowers
of Primula veris from Epirus, Greece. The aim was to investigate the chemical content of the over-harvested
P. veris populations of Epirus and to develop and optimize an extraction protocol to allow
fast, exhaustive, and repeatable extraction. Qualitative analysis revealed that the
P. veris flowers from Epirus were particularly rich in flavonoids, especially flavonol triglycosides
including derivatives of quercetin, isorhamnetin, and kaempferol. A phytochemical
investigation of a 70% hydromethanolic extract from the flowers afforded a new flavonoid,
namely, isorhamnetin-3-Ο-β-glucopyranosyl-(1 → 2)-β-glucopyranosyl-(1 → 6)-β-glucopyranoside, which is also the main constituent of the flower extracts. Its structure
elucidation was carried out by means of 1D and 2D NMR and mass spectrometry analyses.
The HPLC-PDA method was developed and
validated according to the International Council for Harmonisation guidelines.
Since the main flavonol glycoside of the plant is not commercially available, rutin
was used as a secondary standard and the response correction factor was determined.
Finally, the overall method was validated for precision (% relative standard deviation
ranging between 1.58 and 4.85) and accuracy at three concentration levels. The recovery
ranged between 93.5 and 102.1% with relative standard deviation values < 5%, within
the acceptable limits. The developed assay is fast and simple and will allow for the
quality control of the herbal drug.
Key words
Primula veris
- Primulaceae - Primulae flos - quality control - isorhamnetin-3-
Ο-
β-glucopyranosyl-(1 → 2)-
β-glucopyranosyl-(1 → 6)-
β-glucopyranoside - response correction factor
