高效测定发酵液中透明质酸含量的改良CTAB浊度法

Efficient Quantification of Hyaluronic Acid in Fermentation Broth by Modified CTAB Method

透明质酸(HA)在保健品、化妆品及临床医疗等领域都具有重要应用,发酵法是HA生产的主要方法。发酵液中HA含量的快速、准确测定对于HA的生产具有重要意义。在原始十六烷基三甲基溴化铵(CTAB)浊度法测量HA浓度的基础上,提出了去除HA与乙酸缓冲液混合后的水浴步骤、降低CTAB试剂浓度(2.5 g/L)以及确定HA-CTAB络合反应时间(5 min)的改良CTAB法。进一步研究了发酵液各成分对改良CTAB方法的干扰,结果表明,葡萄糖、阿拉伯糖、D-葡萄糖酸前体等对CTAB浊度法没有干扰,但Mg2+等具有较为明显的干扰。通过冰乙醇沉淀与改良CTAB浊度法的耦合,实现了发酵液中HA含量的高效测定。

Hyaluronic acid（HA）,which was mainly produced by fermentation in industry,had many significant applications in the fields of health products,cosmetics,and clinical treatments etc.The rapid and precise measurement of HA concentration in fermentation broth played an important role in microbial production of HA.Quantitative analysis of HA concentration in fermentation broth was modified on basis of original cetyltrimethylammonium bromide（CTAB） turbidity method.The water bath incubation of HA and acetate buffer was eliminated,the optimal CTAB buffer concentration was reduced to 2.5 g/L and the HA-CTAB complex reaction time was restricted at 5 minutes.The interference of fermentation broth to CTAB measurement was further observed.It is revealed that Mg^2＋ concentration exhibits a significant impact on CTAB results,but glucose,arabinose and D-glucuronic acid not.The ethanol precipitation coupled with the modified CTAB measurement was then proposed to implement the effective quantification of HA in fermentation broth.