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Metabolites 2013, 3(4), 912-930; doi:10.3390/metabo3040912
Article

A Rapid Method for the Extraction and Analysis of Carotenoids and Other Hydrophobic Substances Suitable for Systems Biology Studies with Photosynthetic Bacteria

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Received: 23 August 2013; in revised form: 26 September 2013 / Accepted: 27 September 2013 / Published: 11 October 2013
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Abstract: A simple, rapid, and inexpensive extraction method for carotenoids and other non-polar compounds present in phototrophic bacteria has been developed. The method, which has been extensively tested on the phototrophic purple non-sulphur bacterium Rhodospirillum rubrum, is suitable for extracting large numbers of samples, which is common in systems biology studies, and yields material suitable for subsequent analysis using HPLC and mass spectroscopy. The procedure is particularly suitable for carotenoids and other terpenoids, including quinones, bacteriochlorophyll a and bacteriopheophytin a, and is also useful for the analysis of polar phospholipids. The extraction procedure requires only a single step extraction with a hexane/methanol/water mixture, followed by HPLC using a Spherisorb C18 column, with a mobile phase consisting of acetone-water and a non-linear gradient of 50%–100% acetone. The method was employed for examining the carotenoid composition observed during microaerophilic growth of R. rubrum strains, and was able to determine 18 carotenoids, 4 isoprenoid-quinones, bacteriochlorophyll a and bacteriopheophytin a as well as four different phosphatidylglycerol species of different acyl chain compositions. The analytical procedure was used to examine the dynamics of carotenoid biosynthesis in the major and minor pathways operating simultaneously in a carotenoid biosynthesis mutant of R. rubrum.
Keywords: bacteriochlorophyll; carotenoids; HPLC-MS; Rhodospirillum rubrum; systems biology; photosynthesis; quinones; phospholipids bacteriochlorophyll; carotenoids; HPLC-MS; Rhodospirillum rubrum; systems biology; photosynthesis; quinones; phospholipids
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Bóna-Lovász, J.; Bóna, A.; Ederer, M.; Sawodny, O.; Ghosh, R. A Rapid Method for the Extraction and Analysis of Carotenoids and Other Hydrophobic Substances Suitable for Systems Biology Studies with Photosynthetic Bacteria. Metabolites 2013, 3, 912-930.

AMA Style

Bóna-Lovász J, Bóna A, Ederer M, Sawodny O, Ghosh R. A Rapid Method for the Extraction and Analysis of Carotenoids and Other Hydrophobic Substances Suitable for Systems Biology Studies with Photosynthetic Bacteria. Metabolites. 2013; 3(4):912-930.

Chicago/Turabian Style

Bóna-Lovász, Judit; Bóna, Aron; Ederer, Michael; Sawodny, Oliver; Ghosh, Robin. 2013. "A Rapid Method for the Extraction and Analysis of Carotenoids and Other Hydrophobic Substances Suitable for Systems Biology Studies with Photosynthetic Bacteria." Metabolites 3, no. 4: 912-930.



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