High performance liquid chromatography broth

Beran, M. and Zima, J., Determinations of Monensins A and B in the fermentation broth of Streptomyces cinnamonensis by high performance liquid chromatography, Chromatographia, 35, 206, 1993. 

La Higuera, AH Hunt, JL Occolowitz, JH Gilliam. High performance liquid chromatography (HPLC) of natural products. III. Isolation of new tripeptides from the fermentation broth ofP. chrysogenum. Helv Chim Acta 63 1119-1129, 1980. 

A bacterial fermentation broth from S. rimus sp. paromomycinus was fractionated with high-performance liquid chromatography (HPLC), and the individual fractions were screened against 16S and 16Sc. Two pbL aliquots of each LC fraction was combined with a 15-/rL solution containing 2.5 p.M each 16S and 16Sc in 100 mM ammonium acetate and 33% isopropyl alcohol. This mixture was vortexed and incubated for 60 minutes at room temperature prior... 

Figure 2. High performance liquid chromatography profile of centrifuged whole broth and mycelium extract. Column Waters Assoc. 1-125 protein analysis column (30 cm x 7.8 mm). Mobile phase 0.05 M phosphate buffer, pH 6.9. Flow rate 1.0 ml/min, Detector UV at 210 nm.

Additionally, a variety of analytical equipment and techniques that allow the examination of small- (and micro-) scale microbial cultures and their products have become available. Examples include near infrared and Fourier transform infrared spectroscopy, which offer the ability for in situ detection of specific compounds in fermentation broth [22]. However, sensitivity and the required sample volumes pose serious obstacles that still have to be overcome. Another alternative is offered by sensitive pyrolysis mass spectroscopy, which was demonstrated to be suitable for quantitative analysis of antibiotics in 5-pl aUquots of fermentation broth when combined with multivariate calibration and artificial neural networks [91]. The authors concluded that a throughput of about 12,000 isolates per month could be expected. Furthermore, standard chromatographic methods such as gas chromatography or high-performance liquid chromatography, possibly in combination with mass spectroscopy (MS) for detection, can provide simultaneous quantitative detection of many metabolic products. 

K. Holzhauer-Rieger, W. Zhou, and K. Schiigerl (1990) On-line high-performance liquid chromatography for determination of cephalosporin C and by-products in complex fermentation broth. 

Pandey RC, Toussaint MW. High-performance liquid chromatography and thin-layer chio-matography of anthracycline antibiotics. Separation and idemilicatioit of components of the daunorubicin complex from fermentation broth. J Chtomaiogr 1980 198 407-420. 

Neuss, N., R. D. Miller, C. A. Affolder, W. Nakatsukasa, J. A. Mabe, L. L. Huckstep, N. de la Higuera, A. H. Hunt, J. L. Occolowitz, and J. H. Gilliam High Performance Liquid Chromatography (HPLC) of Natural Products. III. Isolation of New Tripeptides from the Fermentation Broth of P. chrysogenum. Helv. Chim. Acta 63, 1119 (1980). 

Reusability of the Electrode. Reusability of the microbial electrode was examined with the sample solution containing 125 r /mi of phenyl acetyl-7ADCA. The results obtained are shown in Figure 6. The cephalosporin determination was performed several times a day. However, no decrease of the potential difference was observed for a week. This system was applied to the determination of cephalosporins in a fermentation broth. Cephalosporin C in the cephalospolium acremonium broth was determined by the conventional method using high pressure liquid chromatography (HPLC) (13) and the electrochemical method. The results obtained are shown in Table 1. The relative error of the determination by the microbial electrode was within 10 %. Therefore, this system can be used for the continuous determination of cephalosporin in a fermentation broth. 

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