高效液相法测定虾青素的含量

Determination of Astaxanthin by High Performance Liquid Chromatographic Method

目的：建立不同虾青素提取物中皂化后游离虾青素的含量测定方法。方法：不同的虾青素提取物以二氯甲烷-甲醇（体积比1∶3）为皂化溶剂,加入1%KOH甲醇溶液于4℃避光皂化7h,在Diamonsil-C18色谱柱（250mm×4.6mm,5μm）上以甲醇-水（95∶5）为流动相,采用紫外检测器在475nm处测定。结果：通过皂化,虾青素提取物中的虾青素类酯可以很好地转化为游离虾青素。在0.0802~0.4812/μg时峰强度与虾青素的质量浓度呈良好线性（r=0.9993）,平均回收率为98.73%,RSD为1.11%。结论：采用本方法测定了多种市售虾青素制剂,结果表明该方法稳定可靠,可用于虾青素的含量测定。

Objective：To establish a high performance liquid chromatographic （HPLC） method for the determination of free astaxanthin in the different astaxanthin extracts after saponification. Methods ： Astaxanthin extracts were saponified with methylene chloride - meth- anol （ 1 ： 3, by volumn ） and 1% KOH methanol avoiding light for 7 hours at the temperature of 4 ℃. The separation of astaxantin was conducted on a Diamonsil C 18 column （250mm × 4.6mm,5 μm） with the mobile phase consisting of methanol -water（95：5 ）. The de- tection waverlength was 475nm. After saponification, the astaxanthin esters converted to free astaxanthin completely. Result： An excel- lent linearity （r =0. 9993） was obtained among the range of 0. 0802 -0. 4812/μg. The average recovery was 98.73% , and the RSD was 1.11%. Conclusion ： The detection method has been applied in the analysis of selling astaxanthin preparations with its good repeat- ability and sensitivity.