HPLC法测定发酵液中灵菌红素的含量

Determination of Prodigiosin in Fermentation Broth by HPLC

摘要:

灵菌红素是具有由三个吡咯环组成的甲氧基吡咯骨架结构的次级代谢产物，因为具有良好的抗癌活性而受到广泛关注。目前，灵菌红素主要是通过分光光度法进行检测。为实现对发酵液中灵菌红素的快速准确测定，实验采用三氟乙酸酸化的流动相体系，利用高效液相色谱法梯度洗脱检测灵菌红素含量。实验结果显示，在优化后的色谱条件下，乙腈-水梯度洗脱体系可以显著改善灵菌红素的色谱峰形以及洗脱时间，并与杂质达到较好的分离度。色谱条件为：sepax Bio-C18色谱柱（150 mm×4.6 mm, 3 μm）；柱温35 ℃；流速为0.6 mL/min。灵菌红素的线性范围为6~150 μg/mL，检测限为0.0058 μg/mL，加标回收率为95.74~98.41%。实验表明，研究所用的分析方法灵敏度、精密度、重现性和准确度均较好，能够较好的应用于灵菌红素发酵样品的分析检测，。

Abstract:

Prodigiosin is a multifaceted secondary metabolite with tripyrrole structure, which attracts great interest for its anticancer activities. At present, the main detection method is spectrophotometry. In order to detect prodigiosinin fermentation broth accurately and rapidly， a high performance liquid chromatography（HPLC）method with gradient elution was developed. The best conditions were established as follows: the separation performed on sepax Bio-C18 column (150 mm×4.6 mm, 3 μm) at 35 ℃ with flow rate of 0.6 mL/min. Under the chosen conditions, prodigiosin could be separated well and showed a symmetrical peak shape with acetonitrile-water gradient elution system. The liner range of prodiogisin was 6~150 μg/mL, and the limit of detection was 0.0058 μg/mL. The recovery of standard addition was 95.74~98.41% with precision of 0.92% relative standard deviation (RSD). The proposed HPLC method could be used for the determination of prodigiosinin fermentation broth with high sensitivity, good repeatability and accuracy.