Quantitative analysis of boeravinones in the roots of Boerhaavia Diffusa by UPLC/PDA.

Introduction: Boerhaavia diffusa is a perennial herb belonging to Nyctaginaceae. Various classes of chemical constituents such as phenolics (boeravinones), terpenoids and organic acids have been reported in B. diffusa roots. As boeravinones have been proposed as putative active constituents for the anti-cancer, spasmolytic and anti-inflammatory activities exhibited by B. diffusa extracts, it is worthwhile developing and validating an ultra-performance liquid chromatography (UPLC) method for analysis of boeravinones in B. diffusa roots.

Objective: To develop and validate a simple, accurate, robust and rapid UPLC analytical method for quality control of B. diffusa roots.

Methods: Samples for analysis were prepared by refluxing powdered root material with methanol for 2 h. The extracts were concentrated, dried and stored at -20°C until their use. A UPLC with photodiode array (PDA) method was developed and validated for the quantification of boeravinones in the roots of B. diffusa. The separation of boeravinones was achieved using a BEH Shield C18 -column (2.1 × 100 mm, 1.7 µm) with gradient elution of methanol and water (0.1% acetic acid), at a flow rate of 0.4 mL/min and detection was carried out at λmax 273 nm.

Results: The UPLC method developed showed good linearity (r(2)  ≥ 0.9999), accuracy and precision.

Conclusion: The UPLC method developed provided a selective, sensitive and rapid analytical method for the quantification of boeravinones in B. diffusa roots. All the validation parameters were found to be within the permissible limits as per International Conference on Harmonisation guidelines.