Research On Extraction、Separation And Irradiation Stability Of Capsanthin

Capsantin, as a pigment, is widely spread in nature, not only with bright color、 strong tinting strength, but also with varieties of bio-activities. Capsantin is a main source of carotenoid, as its importance and shortage in China, so the research on characteristics and separation、purification of capsanthin is urgent necessary. Meanwhile, capsathin is unstable when applied in food industry, in processing and irradiate storage, the color will fade, sometimes it will fade away completely in extreme situations. This paper is using chili powder as raw material, using ultrasonic along-with microwave methods on capsanthin extraction, then using sillicagel column for separation and purification then get relatively high-purity capsanthin,. After irradiation, using HPLC-MC method for analysis, searching for irradiation stability. The main results are as follows:1. Ultrasonic and microwave with solvent extract pepper pigments,we can get the following conclusions:(1) the red pepper powder mixted with ethyl acetate, acetone, ethanol petroleum ether, hexane mixture by1:5, at50℃,250W ultrasonic of30min, the ethyl acetate has the best effect3.5%.(2) single factor experiment and orthogonal experiment results show that, ultrasonic with solvent extract pepper pigments, Influence factors from high to low were:ultrasonic power, ultrasonic temperature, ultrasonic time, ratio of material to liquid, the best combination was A2B2C1D2, namely:ultrasonic temperature50℃, ultrasonic time30min, ultrasonic power190W, ratio of solid to liquid was1:8, with this condition,pepper pigments can reach4%.(3) Ultrasonic and microwave with solvent extract pepper pigments, pepper pigments can be increased by0.78%, the optimum process conditions of capsanthin ultrasonic and microwave with solvent extraction were:ultrasonic temperature50℃, ultrasonic time30min, ultrasonic power190W, the ratio of material to liquid is1:8, microwave power500W, microwave time10min. Under this condition, pepper pigments extraction rate was4.78%.2. After saponification, pepper pigments are separated by silica gel column chromatography and thin layer chromatography:(1) Through the evaluation of pepper pigment color value in single factor test of pepper pigments’saponification, the optimum conditions of saponification:ethanol concentration was50%, the concentration of NaOH was15%, saponification temperature50℃, saponification time2.5h, the ratio of material to liquid5:3.(2) Chilli pigments was purified by silica gel column chromatography, the best condition was selected:silica gel was30g, column diameter15×300mm, elution flow rate300ml/min, petroleum ether and acetone volume ratio10:1. Under this condition, Saponified pepper red pigment first with n-hexane gets yellow eluate, then with petroleum ether and acetone (10:3) gets the red eluate, and finally eluted with65%ethanol gets a pale yellow eluate. According to HPLC-MS analysis, the n-hexane yellow eluate exists β-carotene, petroleum ether and acetone eluate exists violaxanthin diesters, β-carotene and (3-cryptoxanthin,65%ethanol eluate exists capsaicin and capsanthin. But from the HPLC-MS results, each eluent with pepper pigments was mixture, we need further separation.(3) Chilli pigments was purified by thin layer chromatography with different petroleum ether and acetone ratio as developing agent, the best separation effect of chilli pigment was etroleum ether and acetone10:1, which can separate12clear different color spots, its color include red, yellow or between yellow and red. From the origin to the top of the Rf values were0.03,0.1,0.22,0.27,0.33,0.38,0.53,0.58,0.65,0.8,0.87,0.95, Each point analysed by HPLC-MS, we get the following results:Rf0.03spot mainly contains β-cryptoxanthin, Rf0.1spot mainly contains violaxanthin diester, capsanthin monoester, Rf0.22spot mainly contains capsanthin, Rf0.27spot mainly contains capsanthin ester, Rf0.33spot mainly contains capsanthin ester, Rf0.38spot mainly contains capsorubin diester, capsanthin esters, Rf0.53spot mainly contains lutein esters, capsanthin esters, Rf0.58spot mainly contains lutein esters, capsanthin esters, Rf0.65spot mainly contains zeaxanthin diester, Rf0.8spot mainly contains β-cryptoxanthin esters, leaf flavin esters, Rf0.87spot mainly contains lutein, Rf0.95spot mainly contains β-carotene. From HPLC-MS results, the substance is not a single component of the spot, if we want to determine the material composition of each spot, it need further separation and analysis. 3. Pepper pigments of each component irradiated by different doses, the color will change, and with increasing radiation dose, discoloration also intensified. But esterification process can improve pigments’radiation stability. Red pigments’ radiation stability is better than the yellow pigment.Capsanthin has changed partly under2kGy radiation dose, while capsanthin by4kGy,6kGy,8kGy irradiation dose has been completely changed, the capsanthin monomer to radiation is very unstable, so in the study of pure pepper red irradiation dose to the less than2kGy.The results show that to red peppers or red spicy food good color after irradiation, irradiation dose should be controlled in4kGy.