自然科学版
陕西师范大学学报(自然科学版)
功能化合物及其生物活性研究专题
醋酸菌辅酶Q10的RP-HPLC测定方法及提取工艺优化
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颜统晶,张宝善*,韦露莎,沈洋洋
(陕西师范大学 食品工程与营养科学学院,陕西 西安 710119)
张宝善,男,教授,硕士生导师,研究方向为食品微生物及果蔬加工。E-mail:baoshanz@snnu.edu.cn
摘要:
建立了一种测定辅酶Q10(CoQ10)含量的反相高效液相色谱方法(RP-HPLC)并优化了其提取工艺。色谱测定使用Diamonsil反相C18柱(4.6 mm×250 mm,5 μm),流动相甲醇与乙醇的体积比为20∶80,检测波长275 nm,柱温35 ℃,进样体积20 μL,流速1.0 mL/min。在此条件下,CoQ10在1~100 μg/mL质量浓度范围内的线性关系良好(R2=0.999 2),平均回收率为97.07%,相对标准偏差(RSD)为2.93%,可很好地应用于醋酸菌CoQ10含量的测定。在单因素实验的基础上,采用响应面分析法,建立了巴氏醋杆菌中CoQ10酸热法提取的二次多项式回归方程,确定了CoQ10的最优提取工艺:热处理温度为91 ℃,HCl溶液物质的量浓度为0.08 mol/L,热处理时间为38 min。采用该工艺提取的巴氏醋杆菌样品中CoQ10的质量分数为114.81 μg/g,与理论值115.66 μg/g相近,相同样品的CoQ10提取值为优化前的368.83%,说明优化后的提取工艺可以有效提取醋酸菌中CoQ10。
关键词:
醋酸菌;辅酶Q10;反相高效液相色谱法;响应面分析法;提取工艺
收稿日期:
2018-12-29
中图分类号:
TS261.9
文献标识码:
A
文章编号:
1672-4291(2020)05-0034-08
基金项目:
西安市科技计划项目(20193044YF032NS032);陕西省科技计划项目(2019NY-132,2019CGXNG-036)
Doi:
RP-HPLC determination and extraction optimization of CoQ10 in acetic acid bacteria
YAN Tongjing, ZHANG Baoshan*, WEI Lusha, SHEN Yangyang
(School of Food Engineering and Nutritional Science, Shaanxi Normal University,Xi′an 710119, Shaanxi, China)
Abstract:
Coenzyme Q10 (CoQ10) is one of the most important evaluation index for the metabolism ability of acetic acid in acetic acid bacteria. In this investigation, a reversed-phase high performance liquid chromatography (RP-HPLC) method was established to determine the content of CoQ10 in acetic acid bacteria. It was performed on a Diamonsil C18 column (4.6 mm×250 mm, 5 μm) with column temperature setting at 35 ℃. Mobile phase was composed with methanol and ethanol in a volume ratio of 20∶80. The flow rate was 1.0 mL/min and the detective wavelength was 275 nm. Under these conditions, the calibration curve of CoQ10 demonstrated good linear relationship in the range of 1~100 μg/mL (R2=0.999 2) with the relative standard deviation (RSD) of 2.93% and the average recovery rate of 97.07% , proving the effectiveness of this method in determining CoQ10 content in acetic acid bacteria. On the basis of single factor experiments, a quadratic polynomial rgression equation of CoQ10 extraction in acetic acid bacteria was constructed using the response surface methodology. The optimal extraction conditions were established as follow: the temperature was 91 ℃, the concentration of hydrochloric acid was 0.08 mol/L, the extraction time was 38 min. In these optimized conditions, the content of CoQ10 extracted from acetic acid bacteria (114.81 μg/g) was close to the theoretical value (115.66 μg/g), and was 368.83% higher than the extracted value before optimization. The results can be used to further utilize acetic acid bacteria and develop functional CoQ10 products.
KeyWords:
acetic acid bacteria;coenzyme Q10;RP-HPLC;response surface methodology;extraction technology