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The protonated form of bromothymol blue has its peak absorption at 427 nm thus transmitting yellow light in acidic solutions, while the deprotonated form has its peak absorption at 602 nm thus transmitting blue light in more basic solutions. [3] In contrast, highly acidic bromothymol blue is magenta in color. The general carbon skeleton of ...
It was developed in 1953 by Hugh and Leifson to be utilized in microbiology to determine the way a microorganism metabolizes a carbohydrate such as glucose (dextrose). [1] OF-glucose deeps contain glucose as a carbohydrate, peptones, bromothymol blue indicator for Hugh-Leifson's OF medium or phenol red for King's OF medium, and 0.5% agar.
Universal indicator components Indicator Low pH colour Transition pH range High pH colour Thymol blue (first transition) Red 1.2 – 2.8 Yellow Methyl orange: Red 3.2 – 4.4 Yellow Methyl red: Red 4.8 – 6.0 Yellow Bromothymol blue: Yellow 6.0 – 7.6 Blue Thymol blue (second transition) Yellow 8.0 – 9.6 Blue Phenolphthalein: Colourless
Bromothymol blue was added in order to reduce false positives. The citrate agar is green before inoculation, and turns blue, because of BTB as a positive test indicator, meaning citrate is utilized. The test is also prepared on a slant to maximize bacterial growth for an even better indication of the use of citrate.
Bromothymol blue is the indicator used in the agar, it changes to yellow in case of acid production during fermentation of lactose or changes to deep blue in case of alkalinization. Lactose-positive bacteria build yellow colonies. Bacteria which decarboxylate L-cystine cause an alkaline reaction and build deep blue colonies. [1]
Bacteria are inoculated on a medium containing sodium citrate and a pH indicator such as bromothymol blue. The medium also contains inorganic ammonium salts, which are utilized as sole source of nitrogen. Use of citrate involves the enzyme citrate lyase, which breaks down citrate to oxaloacetate and acetate.
The increase in pH then causes color change in the bromothymol blue indicator, turning it blue. Under neutral conditions the medium remains a green color. The color change to blue is useful because growth on Simmons' citrate agar is often limited and would be hard to observe if it were not for the color change.
The standards used include potassium dichromate (isosbestic points at 339 and 445 nm), bromothymol blue (325 and 498 nm) and congo red (541 nm). The wavelength of the isosbestic point determined does not depend on the concentration of the substance used, and so it becomes a very reliable reference.
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