| CrocetinⅠis clinically used to treat diabetes,obesity,Alzheimer’s disease,atherosclerosis and other diseases.It was extracted from the stigma of crocus in Iridaceae.High purity CrocetinⅠcan not only provide material guarantee for the accuracy and traceability of CrocetinⅠanalysis results,but also enrich the reference material database of China.In this paper,the purification technology and antioxidant activity of CrocetinⅠwere studied.Firstly,macroporous adsorption resin technology was used to purify CrocetinⅠ,and AB-8 was selected as the best resin by static adsorption and desorption experiments.The experimental conditions of AB-8 resin dynamic analysis were determined by optimizing the concentration of sample solution,the volume fraction of eluent,the flow rate of sample,the flow rate of eluent and the amount of eluent.HPLC analysis showed that the purity of CrocetinⅠincreased to 76.70% after AB-8 resin purification.Silica gel column chromatography was used to further purify CrocetinⅠ,and the effects of different factors were investigated.The optimal operating conditions were determined as follows: the sample loading was 0.3 g,the eluant type was methanol chloroform gradient elution,the ratio of column diameter to height was 1:7,and the elution flow rate was 2.0m L/min.Under this condition,the purity of CrocetinⅠreached 98.51%.The yield was60.78%.At the same time,the preparation chromatography technology was used to purify CrocetinⅠ,and the best preparation conditions were obtained: the mobile phase was methanol water isoelution,the detection wavelength was 262 nm / 327 nm,the sample volume was 2.0 m L,the flow rate was 15 m L / min,the purity of CrocetinⅠincreased to99.52%;and the stability of CrocetinⅠwas studied.The purified products were qualitatively analyzed by mass spectrometry,infrared spectroscopy and nuclear magnetic resonance spectroscopy.Finally,the antioxidant capacity of CrocetinⅠwas evaluated by salicylic acid method and DPPH method,and compared with that of vitamin C. |
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