坛紫菜氨基酸提取工艺影响条件的优化与分析

Optimization and analysis of Porphyra haitanensis amino acid extraction methods

以坛紫菜(Porphyra haitanensis)为实验材料,采用盐酸水解法和浸提法,以日立L-8900型高速全自动氨基酸分析仪为测定仪器,分别研究了坛紫菜总氨基酸(TAA)和游离氨基酸(FAA)在提取过程中各条件的不同水平对提取结果的影响.实验结果表明:(1)利用正交设计L9(34)对影响TAA提取的4个条件(藻体质量、稀释倍数、干燥温度、空白值)在3个水平上进行优化实验,藻体质量、稀释倍数对测定结果影响显著,干燥温度影响不显著;通过实验筛选出各条件的最佳水平为:藻体质量为0.050 0g,稀释3倍,干燥温度为60℃.(2)利用正交设计L16(45)对影响FAA测定的5个条件(藻体质量、提取液、提取温度、提取时间、空白值)在4个水平上进行优化实验,结果表明藻体质量、提取液、提取温度、提取时间的不同水平对测定结果均有显著影响,其中藻体质量对结果的影响最大;通过实验筛选出各条件的最佳水平为:藻体质量为0.010 0 g,提取液为去离子水,提取温度为60℃,提取时间为4 h.

Total amino acid and free amino acid of Porphyra haitanensis extracted by HCl-Hydrolysis and soaking were analyzed using a Hitachi L-8900 Amino Acid Analyzer under different extraction conditions,i.e.with different levels of each factor.The results showed that（1） when using orthogonal experimental design L9（34） to design different optimization experiments to evaluate four factors（weight of thallus,dilution ratio,temperature and blank） that affect the extraction of total amino acid in three levels,the results were significantly affected by the weight of the thallus and extractive solution,and insignificantly affected by dry temperature.The optimum conditions were found to be as follows： 0.050 0 g thallus,three times dilution and drying at 60℃.（2） when using orthogonal experimental design L16（45） to develop different optimization experiments based on five factors（weight of thallus,dilution ratio,temperature,extraction time and blank） that affect the extraction of free amino acid,the results were significantly influenced by the five factors,with the weight of thallus having the greatest effect.The best conditions were as follows： 0.010 0 g thallus,deionized water,extraction at 60℃ for 4 h.These optimization experiments to evaluate the extraction methods of Porphyra haitanensis provide a good foundation for its precise measurement and support for the breeding of high-quality Porphyra haitanensis.