UPLC法测定糕点中β-胡萝卜素的含量

Determination of Beta Carotene in Pastries by UPLC

为获得糕点中β-胡萝卜素的定性定量分析的方法,以添加了β-胡萝卜素的糕点为原材料,通过对不同提取试剂、有无皂化反应等试验条件的筛选,优化糕点中β-胡萝卜素的提取条件;通过确定色谱条件,以及重复性、精密度、回收率试验,建立糕点中β-胡萝卜素的超高效液相色谱法分析体系。结果表明,以正己烷作为提取剂,经过皂化反应提取糕点中的β-胡萝卜素,同时采用Agilent Proshell SB-C18色谱柱,以乙腈︰甲醇=85︰15为流动相,在流速0.5 mL/min、最大吸收波长450 nm的条件下进行测定,是糕点中β-胡萝卜素定性定量分析的最佳方法。该方法检出限为0.72μg/100 g,在0.2~4.0μg/mL质量浓度范围内线性关系良好,线性相关系数为0.999 7。当加标含量分别为2.8, 28.0和10 000.0μg/100 g时,加标回收率分别为86.4%, 88.4%和84.6%。连续进行7次试验,相对标准偏差小于6.97%。该方法所使用有机试剂种类少、用量小,具有对人体危害小、容易操作、稳定好的优点。样品经过皂化反应,用正己烷提取与超高效液相色谱法结合的分析体系适合糕点中β-胡萝卜素的定性定量分析。

In order to obtain the qualitative and quantitative analysis method of β-carotene in pastries, pastries with beta-carotene have been used as research materials. Different extraction agents, saponification or not and other experimental conditions are studied. The extraction condition of β-carotene from pastry is optimized. Chromatographic conditions, repeatability, precision and recovery tests are determined. The ultra-performance liquid chromatography(UPLC) is established for the determination of β-carotene in pastries. The results show that the best extract solvent for β-carotene in pastries is normal hexane. It shall be saponified and analyzed by the column of Agilent Proshell SB-C18 with acetonitrile: methanol=85︰15 as mobile phase at 450 nm. Meanwhile, the flow rate is 0.5 mL/min. The detection limit of this method is 0.72 μg/100 g. And the correlation coefficient is 0.999 7 in the range of 0.2 μg/mL to 4.0 μg/mL. Respectively, the recoveries are 86.4%, 88.4% and 84.6% when the spiked concentrations are 2.8, 28.0 and 10 000.0 μg/100 g. The relative standard deviation of 7 experiments is less than 6.97%. The type and amount of organic reagents used in this method are relatively small. Moreover, it has the advantages of less harm to human body, easier operation and better stability. The analysis system has been combined with normal hexane extraction and UHPLC. Furthermore, it is suitable for qualitative and quantitative analysis of β-carotene in pastries.