Phytochemical Profile Analysis by LC-ESI-MS n and LC-HRESIMS validated HPLC-method for quantification of rutin in herbal drug and products from leaves of Croton blanchetianus.

Phytochemical analysis of Croton blanchetianus leaves was performed by Liquid Chromatography-Electrospray Ionization-Mass spectrometry. After that, a High Performance Liquid Chromatography method was developed and validated for the determination of rutin in herbal drug and products of C. blanchetianus. The separation was achieved on a C 18 column, and the mobile phase was composed of ultrapure water and methanol (acidified with trifluoracetic acid) with a gradient at 0.8 ml/min. The method was validated following international guidelines. The chemical analysis revealed the presence of flavonoids, among them rutin was used as the standard for validation. In the HPLC the presence of rutin was observed at 24.7 min. The method was robust, with no significant variations, and linear in the range evaluated with R 2 >0.99. In the matrix effect, it was possible to prove the absence of interference of the constituents in the herbal drug. The precision was determined with a relative standard deviation<1.34%. The recovery results were achieved between 89.29 and 101.21%. Furthermore, with partial validation, the suitability of the method was proven to be suitable for the liquid extract, dry extract, and effervescent granules. So, this study demonstrated that the method is effective for quality control analysis of C. blanchetianus leaves and products.