紫荆花色素的提取、粒度分析和毒性检测

Extraction,Analysis of Particle Size and Toxicity Detection of Flower Pigment from Cercis Chinensis

鲜花色素是一种优质的天然食用色素。试验以产率和色价评价5种不同提取方法获得紫荆花色素,并对喷雾干燥后的色素颗粒进行粒径和粒度分布分析,以CCK-8试剂盒检测紫荆花色素对乳腺癌MDA-MB-231细胞活力的影响,按国标方法检测鲜花色素对小鼠的急性致死性毒性。结果显示:纯水浸提盛花期紫荆鲜花色素的色价最高,以95%乙醇浸提末花期紫荆鲜花的色素产率最高。检测喷雾干燥后的色素颗粒,以纯水提取紫荆花色素颗粒粒径小(2~10μm)、颗粒形态规范(体积平均粒径与表面积平均粒径差值小),而亚临界丁烷提取的紫荆鲜花色素颗粒粒径偏大(5~20μm),颗粒间形态差异较大。紫荆花色素喷雾形成的颗粒粒径集中在5~10μm之间,粒度分布比较均匀便于制剂。CCK-8检测细胞毒性发现,以95%乙醇浸提末花期紫荆鲜花的色素对乳腺癌MDA-MB-231细胞活力有一定影响,当色素质量浓度大于300μg·mL^(-1)时,细胞活性维持在12.3%左右,当质量浓度在100μg·mL^(-1)以下时,细胞活性可保持在80%以上。通过国标法检测该色素对小鼠的急性致死性毒性发现,小鼠的最大耐受剂量(MTD)大于15000 mg·kg-1体质量,因此判定提取的紫荆花色素的急性经口致死性毒性为无毒级别。结论:首次检测了紫荆花色素的喷雾颗粒粒径及其分布,并检测了紫荆花色素的细胞毒性和急性毒性,这些结果为紫荆花色素的生产和应用提供了基础数据。

Flower pigment is a kind of high quality natural edible pigment.In this paper,we extracted flower pigment from Cercis chinensis with five different methods evaluated by the yield and color value.Then,the particle size and size distribution of pigment particles after spray drying were analyzed.The toxic effect of flower pigment on the viability of MDA-MB-231 cells was explored by CCK-8 test kit.To determine the acute lethal toxicity of flower pigment,flower pigment samples were gavaged to mice according to the national standard method.The results showed that the color value of flower pigment extracted by pure water with fresh flower was the highest among five different methods,and the yield of flower pigment extracted by 95%ethanol with withered flower was the highest.Detection of pigment particles after sprayed drying,the particle size of pigment extracted by pure water is smaller(2-10μm),particle shape is more regular,i.e.the difference between volume average particle size and surface area average size is small.As to the pigment particle extracted by subcritical butane extraction with fresh flower,particle size is larger(5-20μm),the morphological difference of particles is relatively large.The particle size of the pigment spray formed concentrated between 5-10μm,particle size distribution is relatively uniform and easy preparation.The results of CCK-8 assay showed that the pigment extracted from 95%ethanol with withered flower affected the viability of breast cancer MDA-MB-231 cells.When the concentration of pigment was above 300μg·mL^(-1),the cell viability decreased to 12.3%,and the concentration lowered to 100μg·mL^(-1),the cell viability increased to 80%.The acute oral lethal toxicity of the pigment extracted 95%ethanol with withered flower was considered to be non-toxic because the maximum tolerated dose(MTD)of the pigment in mice was more than 15000 ng·kg-1body weight.In this study,the particle sizes and distributions of pigment were detected for the first time,at the same time,the cytotoxicity and acute toxicity of the pigment were tested.These results provide basic data for the production and application of the flower pigment from Cercis chinensis.