Roses Total Procyanidins Separation, Purification And Analysis

This paper is about the extraction , purification and analysis of the proanthocyanins from Rosa rugosa .The condition of the extraction of proanthocyanins from Rosa rugosa was studied . The TLC was used to analysed the proanthocyanins from Rosa rugosa qualititively and the UV was used to analysed quantitatively the proanthocyanins from Rosa rugosa primarily. Then macroporousadsorption resin was used to purify the Proanthocyanidins from Rosa rugosa. Finally the HPLC was used to analyse quantitively the proanthocyanins from Rosa rugosa precisely.In the experiment, The dried rose power was used as material, ethanol was used as solution to extract proanthocyanidins from Rosa rugosa and some factors ,including extracting temperature , the ratio between the material and the extracting solution , the concentration of ethanol , extracting time were studied. The result shows that The best extracting method is : the ratio between the extracting solution and the material: 7:1 , extracting temperature: 50℃, extracting time: 45min, the concentration of ethanol: 70%. The ratio of extraction is 42.58%.After the qualitative analysis of the proanthocyanins from Rosa rugosa, the proanthcyanins was determined to exist according to the result of the color showed. Then three familiar macroporousadsorption resins : AB-8, D-152, D1300 were choosed to purify the extract of proanthocyanins from Rosa rugosa . The result after purification shows the ratio of extraction and purification is 2.54% and 72.70% respectively after being purified by AB-8 macroporousadsorption resins and it is the highest in three macroporousadsorption resins.This result shows that AB-8 macroporousadsorption resin is the fittest one to purify the proanthocyanins from Rosa rugosa.Finally the proanhtocyanins B₂ was used to be the standard to annalyse the proanthocyanins from Rosa rugosa qualitatively and quantitively by HPLC. Analyses were performed on Waters C₁₈ 5μm,4.6 × 150mm , at a flow rate of 1ml/min, using a 20 μl injection volume, detection at 280nm. The elutionprogramme is as follows: A: formic acid :water=l:9(v/v),B: formic acid :water :MeOH=l:4:5(v/v/v);0⁴min, 90% A, 10% B;4²1min, 10% 100% B;21²5min, 100% B;25²6min, 100% 10% B. The result shows that the proanthocyanins from Rosa rugosa has the proanthocyanin B2. The result of the methodology of the total proanthocyanins from Rosa rugosa is as follows:the standard line : Y=1279.9X+3041.1, R=0.9995,the analytical response was linear (R=0.9995)between 27.5 ug/ml 220 ug/ml of proanthocyanins. The reproducibility and the limit of detection were 99. 0%¹01.4% and 3.4 lig/ml.It shows that it is reasonable to use HPLC to analyse the total proanthocyanins from Rosa rugosa qualitively and quantitively.