HPLC測定黑果枸杞鮮果中花青素含量的方法研究

束彤 于瑤 馬明芳 陳睿

摘? 要：目的：優(yōu)化并建立黑果枸杞鮮果中HPLC的測定方法。方法：采用乙醇、鹽酸、水為提取液，超聲提取、水浴水解后用HPLC測定黑果枸杞鮮果中的花青素含量。對乙醇濃度、鹽酸濃度、超聲時間、水浴溫度進行正交實驗條件優(yōu)化，對實驗的檢出限、線性關(guān)系、重復(fù)性及回收率進行了考察。結(jié)果：確定各因素影響主次順序為水解溫度>鹽酸濃度>超聲時間>乙醇濃度，最優(yōu)提取條件為鹽酸30%、乙醇50%、超聲時間15min、沸水浴。本方法6種花青素的檢出限都可達到12mg/kg，在1～50μg/mL范圍內(nèi)，除天竺葵色素R2=0.9989外，其余花青素R2均大于0.999，6次獨立平行測定RSD均小于4%，6種花青素回收率在89.0%～91.7%。結(jié)論：該方法可以用于黑果枸杞鮮果中花青素含量的測定。

關(guān)鍵詞：黑果枸杞鮮果;花青素;高效液相色譜;正交實驗

中圖分類號 TS264.4文獻標識碼 A文章編號 1007-7731（2019）01-0024-07

Determination of Anthocyanin in Fresh Fruit of Lycium ruthenicum Murr.by HPLC

Shu Tong et al.

（Qinghai Province Institute for Food Control，Xining 810000，China）

Abstract：Objective：A high performance liquid chromatographic （HPLC） method was established for the determination of anthocyanin in fresh fruit of Lycium ruthenicum Murr.，and the orthogonal method was use to research the optimum extraction conditions. Methods：Anthocyanin were extracted by ultrasonic （solvent extraction were ethanol，hydrochloric acid and water），extract was hydrolyzed into major anthocyanins in hot water bath，and then HPLC was used to test the content of anthocyanins. Four factors （concentration of ethanol，concentration of hydrochloric acid，ultrasonic time and water bath temperature） were optimized by orthogonal experiment. The limits of detection （LOD），linearity，repeatability and recovery of the method were studied. Result：The order of primary and secondary of extract factor were temperature of water bath，concentration of hydrochloric acid，ultrasonic time and concentration of ethanol，and optimal extraction conditions were 30% hydrochloric acid，50% ethanol，ultrasonic extract 15 min and boiling water bath;and the result showed that LOD could up to 12mg/kg of all 6 kinds of anthocyanins，the linearity of the method is good （R2 of pelargonidin was 0.9989，and the others'were all above 0.999，over the concentration range from 1 μg/mL to 50μg/mL ），the relative standard deviations （RSD） of 6 tests were all less than 4%，and the ranges of recoveries at different levels were 89.0%～91.7%. Conclusion：the method can be used to test anthocyanidins in fresh fruit of Lycium ruthenicum Murr..

Key words：Fresh fruit of Lycium ruthenicum Murr.;Anthocyanin;High performance liquid chromatographic （HPLC）;Orthogonal experiment

花青素，又名花色素，是一類水溶性的類黃酮化合物，廣泛存在于植物的花、果實、莖、葉中[1]?；ㄇ嗨鼐哂卸喾N生理功能，包括抗氧化、抗輻射、抗衰老以及調(diào)節(jié)血脂功能，能有效防治動脈粥樣硬化等慢性疾病[2-4]。本試驗對黑果枸杞鮮果中花青素的提取試劑、提取條件、色譜條件等進行了研究，以期建立黑果枸杞鮮果中花青素成分及含量測定的高效液相色譜法，為黑果枸杞鮮果中花青素的檢測提供技術(shù)支撐，也為黑果枸杞鮮果及下游產(chǎn)品的質(zhì)量標準檢測體系提供科學(xué)依據(jù)。