Quantitative Analysis of Quality Control of Natural Medicine by $^1H-NMR$ Spectrometry-Quantitative Analysis of Hesperidin from Citrus unshiu

Objectives : In this paper, we describe that spectroscopy may be superior to the conventional HPLC for the quantitative analysis of hesperidin from Citrus unshiu. Methods : spectra (400 MHz) were recorded in using a Varian UNITY Inova AS 400 FT NMR spectrometer. One hundred milligram of powdered Citrus unshiu was weighed out and mixed with 1 ml of with sonication for 30 min (room temperature). The extracts were filtrated through a 0.45 PVDF filter and 0.5 ml of filtrated extract used for quantitative measurement (added 1 mg of dimethyl terephthalate as internal standard). The quantity of hesperidin was calculated by the ratio of the intensity of the compound to the known amount of internal standard. For HPLC analysis, the half gram of plant material was extracted with 60 ml of MeOH for 2 hours. The extracts were made 100 ml volume and analyzed by a Waters HPLC system using a YMC ODS column. The total flow rate was 1.0 ml/min with a sample volume 10 and UV detection at 280nm. Results : The contents of hesperidin in Citrus unshiu was determined % in the quantitative method and in HPLC method. Using the quantitative the contents of hesperidin can be determined in much shorter time than the conventional HPLC measurements. Conclusions : From those results, the advantages of quantitative analysis are that can be analyzed to identify and quantify, and no reference compounds required for calibration curve. Besides, it allows rapid and simple quantification for hesperidin with an analysis time for only 10 min without any pre-purification steps.