应用真空紫外线探测器用于液体色谱:在发酵过程中同时分析四种有机酸和四种碳水化合物
Mayur Bansal1, Annika Dombrowski2, Chinoso Nwanebi3
1Interdisciplinary Life Sciences Graduate Program, The University of Texas at Austin, 2500 Speedway Avenue, Austin, TX 78712, United States.
概括
一个新的真空紫外线探测器用于高性能液体染色学准确量化有机酸和碳水化合物在发酵. 这种方法提供了扩展的动态范围,简化了生物处理应用的样本准备.
科学领域:
- 分析化学 分析化学
- 生物技术是生物技术.
- 生物化学 生物化学
背景情况:
- 生物处理中的代谢物分析对于监测发酵至关重要.
- 像折射率这样的传统探测器在动态范围和选择性方面存在局限性.
- 真空紫外线 (VUV) 检测提供了改善代谢物量化的潜力.
研究的目的:
- 为了验证和描述一个原型的VUV吸收探测器系统,用于高性能液态染色学 (HPLC).
- 评估探测器对有机酸和碳水化合物的同时量化能力.
- 评估VUV探测器在线性,动态范围和复杂生物样品的光谱选择性方面的性能.
主要方法:
- 使用原型VUV探测器与HPLC相结合,同时分析八种分析物 (四种有机酸和四种碳水化合物).
- 采用阿米内克斯HPX-87H柱,使用硫酸移动相和样品预热.
- 收集了七个离散波长波段的全频谱VUV数据,用于光谱解卷.
主要成果:
- 在0.0228-3.59g/L和高线性 (R2 = 0.941-0.998) 之间达到检测极限.
- 显示了延长的线性反应高达100g/L,明显比折射率检测更广泛.
- 通过光谱解卷,成功地分离和量化了共同排泄的峰值,并准确地回收了有机酸和糖.
- 在Saccharomyces cerevisiae发酵中监测葡萄糖和乳酸的生产,展示实际应用.
结论:
- LC-VUV系统提供了强大的,高通量代谢物分析,具有出色的定量准确性和光谱选择性.
- 扩展的动态范围通过减少样品稀释需求来简化工作流程.
- 尽管VUV探测器在非常低度下敏感性较低,但它显示出生物处理应用的巨大潜力,特别是对于高度样品.
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