Veremchuk O.A., Moiseyev D.V.
Validation of the technique of flavonoids quantitative determination in heather shoots
Education establishment "Vitebsk State Order of Peoples’ Friendship Medical University"
Abstract.
The objective of the present research work was to develop and validate the technique for quantitative determination of flavonoids on dry material in heather shoots by liquid chromatography method.
The chromatographic separation was performed on a Zorbax SB C8 column (4,6T250 mm, 5 µm). The mobile phase comprised phosphate buffer solution pH=3,0 and acetonitrile (80:20 v/v).
Validation of the developed technique of flavonoids quantitative determination in heather shoots was performed with the following characteristics taken into account: specificity, linearity, accuracy, precision (repeatability and intermediate precision) and robustness. The detector response (peak area for isoquercitrin) was linear for concentrations ranging from 50 to 2000 µg/ml and correlation coefficient was 0,9995. The accuracy of the developed technique was checked by carrying out the recovery experiment by using the method of «standard addition». The value of percent recovery was found to be 102,1±1,3%. The technique is precise because the values of relative standard deviations proved to be less than 5,0%. Changes in the extraction time (60±5 min), mobile phase composition (±1%) and rate (1,0±0,1 ml/min) had no significant effect on the detector response, thus the technique may be considered robust. The flavonoids content on isoqercitrin and dry material in heather shoots made up 36,8±1,3 mg/g.
Thus the developed and validated technique for quantitative determination of flavonoids in heather shoots is specific, linear, accurate, precise and robust within the range of application, and can be used for routine quality control of heather shoots.
Key words: flavonoids, isoquercitrin, liquid chromatography, heather.
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