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Method development for PPB culture screening, pigment analysis with UPLC-UV-HRMS vs. spectrophotometric methods, and spectral decomposition-based analysis

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Grassino, M., Batstone, D. J., Yong, K. W. L., Capson-Tojo, G. and Hülsen, T. (2022) Method development for PPB culture screening, pigment analysis with UPLC-UV-HRMS vs. spectrophotometric methods, and spectral decomposition-based analysis. Talanta, 246 . p. 123490. ISSN 0039-9140

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Article Link(s): https://doi.org/10.1016/j.talanta.2022.123490

Publisher URL: https://www.sciencedirect.com/science/article/pii/S0039914022002867

Abstract

PPB carotenoids are usually measured through spectrophotometric analysis, measuring total carotenoids (TCs) which has low accuracy and cannot identify individual carotenoids or isomers. Here, we developed an ultra-performance liquid chromatography method with ultraviolet and high-resolution mass spectrometry detection (UPLC-UV-HRMS) to quantify neurosporene, lycopene, and bacteriochlorophyll a contents in PPB cultures. The method exhibited satisfactory recoveries for individual pigments (between 82.1% and 99.5%) and was applied to a range of mixed PPB cultures. The use of a C30 column also enabled the detection of three different isomers of lycopene. In addition, a method for anaerobic photoheterotrophic PPB cultivation to acquire live-cell spectrophotometric information was developed and tested by modifying a standard microbial culture microplate system. A rapid, and relatively low effort principal component analysis (PCA) based decomposition of the whole-cell spectra for pigment analysis in the microplates was also developed. Analysing whole-cell spectra via PCA allowed more accurate prediction of individual pigments compared to absorption methods, and can be done non-destructively, during live-cell growth, but requires calibration for new media and microbial matrices.

Item Type:Article
Business groups:Biosecurity Queensland
Keywords:Purple phototrophic bacteria Pigments Carotenoid Lycopene Bacteriochlorophyll
Subjects:Agriculture > Agriculture (General) > Agricultural chemistry. Agricultural chemicals
Technology > Technology (General) > Spectroscopy
Deposited On:17 May 2022 06:24
Last Modified:17 May 2022 06:24

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