高效液相色谱法测定发酵液中血红素含量
Determination of heme content in fermentation broth by high performance liquid chromatography
史珂欣 1王文雅 1国振 2黄建 3唐艳斌3
作者信息
- 1. 北京化工大学 生命科学与技术学院,北京 100029
- 2. 中国计量科学研究院,北京 100029
- 3. 中国疾病预防控制中心 营养与健康所,北京 100050
- 折叠
摘要
血红素中的卟啉分子在溶液中(特别是在碱性溶液中)会发生聚集而形成多聚体,导致血红素溶液不稳定,从而影响血红素含量测定的准确性.为了解决这一问题,通过在NaOH溶液中加入表面活性剂Triton X-100 构建了血红素的稳定溶液体系,实验结果表明血红素在 Triton X-100/NaOH溶液(含有 25 g/L Trition X-100 和0.1 mol/L NaOH)中稳定性较好,在0～6d内血红素含量变化不显著.采用高效液相色谱法(HPLC)分析了微生物发酵液中血红素含量,得到优化的HPLC测定条件为:以三氟乙酸/甲醇-乙腈体系为流动相进行梯度洗脱,流速为1.0 mL/min,柱温为室温(25～28℃),分离时间为50 min.方法学评价结果显示,所建立的HPLC测定方法的重复性好、准确度高,可用于发酵液中血红素含量测定.
Abstract
Porphyrin molecules in heme aggregate and form polymers in solution(especially in alkaline solution),resulting in instability of heme solutions,thus affecting the accuracy of heme content determination.In order to solve this problem,a stable solution system of heme was constructed by adding surfactant Triton X-100 to NaOH solution.The experimental results showed that heme was stable in Triton X-100/NaOH solution(containing 25 g/L Triton X-100 and 0.1 mol/L NaOH),and the heme content did not change significantly within 0-6 days.The content of heme in microbial fermentation broth was analyzed by high performance liquid chromatography(HPLC).The optimized HPLC conditions were as follows:trifluoroacetic acid/methanol-acetonitrile system was used as the mobile phase for gradient elution,the flow rate was 1.0 mL/min,the column temperature was room temperature(25-28℃),and the separation time was 50 min.The results showed that the established HPLC method had good repeatability and high accuracy,and could be used for the determination of heme content in fer-mentation broth.
关键词
血红素/微生物发酵/高效液相色谱法(HPLC)/含量测定Key words
heme/microbial fermentation/high performance liquid chromatography(HPLC)/content determina-tion引用本文复制引用
基金项目
国家卫生健康委员会微量元素与营养重点实验室开放基金(WLKFZ202204)
出版年
2024
NETL
NSTL
万方数据