Hello,
I will be culturing anaerobic bacteria in broth containing sulfate. I have to measure sulfide produced in broth at different interval. I checked the method of "Cline, J. D. Spectrophotometric determination of hydrogen sulfide in natural waters. Limnol. Oceanogr. 1969, 14, 454−458". This paper seems very complicated to me to use, I could not understand how to use the method in lab. Please suggest me method.
This is very popular method:
Trüper HG, Schlegel HG. (1964) Sulphur metabolism in Thiorhodaceae. Quantitative measurements of growing cells of Chromatium okenii. Antonie van Leeuwenhoek 30: 225-238.
Since many years we measure sulfide with the method of Golboa-Garber (Analytical Biochemistry 43:126, 1971). Recent commercially available test kits for sulfide are also based on this method (see www.merckmillipore.com/GB/en/product/Sulfide-Test,MDA_CHEM-114779 )
You can use a hydrogen sulfide test kit (HACH model HS-WR) for rapid measurements. You have to use it immediately after you collect your samples as hydrogen sulfide volatizes quickly. I use this test previously :
Personna, Y. R., L. Slater, D. Ntarlagiannis, D. Werkema, and Z. Szabo (2013), Complex Resistivity Signatures of Ethanol Biodegradation in Porous Media, J. of contaminant hydrology, 153, 37-50.
Personna, Y. R., D. Ntarlagiannis, L. Slater, N. Yee, M. O’Brien, and S. Hubbard (2008), Spectral Induced Polarization and Electrodic Potential Monitoring of Microbially Mediated Iron Sulfide Transformations, J. Geophys. Res, 113, G02020.
please find attached a paper by Ralph Cord-Ruwish
in which he describes a method for sulfides measurement in anaerobic cultures using CuSO4 in HCl
Method based on Cline 1969 is more complicated to perform but it is very precise. Samples after fixation can be stored for some time before analysis. Method with CuSO4 is very quick and absorbance has to be measured immediately but it is not very precise since formed CuS tends to aggregate and form flocs that disturb measurement of absorbance.
Arvind, please, consider that some sulfide in broth could be formed not from sulfate but released from S-containing organic compounds of broth.
I forgot to add that CuSO4 method can not measure FeS if formed but Cline method can
Thanks researchers
Method by Ralph Cord-Ruwish is easy one and I used this for standard of Sodium Sulfide. Standard curve was not exactly as it should be, but I hope proper care of anaerobic conditions will help. Aim is to measure sulfide produced from sulfate added in broth.
I' don't have experience regarding handling of sodium sulfide. The available box have liquid and took paste (not dry powder), washed with anoxic water, filtered & dried at 50 degree and used further to make standard.
Any more input please..
Dear Arvind,
Very often commercially available Sodium sulfide hydrate is used for standard curve preparation. It is supposed to have following formula: Na2S x 9 H2O, that is used for calculations. Very often is is in the form of krystalls. If the surface is white - then the krystalls are not good. It is recommended indeed to wash/rinse krystalls quickly with anoxic water and dry with a paper. If you obtained colorless transparent krystalls - then you can use them. We never dried krystalls at 50C. Numerous preparation of standard curve gives quite good results.
I hope that helps!
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