I am currently using ion chromatography but I would like to explore other options too. I have also used ascorbic acid using some test kit, but it only detects low levels. Are there other methods and if anyone has used ion chromatography, can I get the protocol.
By using a known dilution of you sample you can use the Ascorbic Acid method. Once your measurement is obtain, using your dilution factor, you can determine the concentration in your sample.
I agree with Raphael! And you can use spectrophotometer to measure phosphates.
When I measured the concentration of phosphates (as P) in the solution (25 mg P/L), I diluted it 100 times. And the measurement I got was 0.38 Abs.
Dear Kamal,
Attached you will find a pdf file where the "standard methods for the examination of water and wastewater" are properly described. Look at page 239 where distinc standardized methods for the analysis of phophorous including sample preparation are defined and described (selection of method, precision and bias, sampling and storage, apparatus, reagents...).
Following one of th standardized methods in the analysis of your ssamples ensures the quality of your analytical procedure and thus of your results.
I hope, this will help you for this an other analysis. The file also contents methods for the analsysis of 1) inorganic nonmetallic contituents, aggregate organic constituents and organic compounds)
Good luck
Dear Kamal,
Phosphate measurement by UV-VIS spectrophotometry (either in the field with a portable equipment or in the lab) works pretty well up to 4-5 mg/L PO4 (or around 1.5 mg/L PO4-P). Thus, in most cases you would not need to dilute your waters by high factors. You have commercially available cuvette tests which are easy to use and there is available QA/QC procedures to ensure the accuracy of your measurements. I obatined good results (comparable to those obtained by ion chromatography) in strongly contaminated mine waters with a complex metal matrix. You may have some problems with possible interferences with certain elements, specially metals (in concentrations of mg/L). Then you may need to use interference supressors (also available in the market). Thus, the precision of this method will depend on the composition of your waste waters.
Cheers,
Thank you everyone for your answers. However, I have to analyze lots of sample on a daily basis. I am looking for method that is easy and efficient and also takes short time. I am using ion chromatography currently. I am looking for other methods so i can compare results. Thank you
Hello Kamal,
For a quickest way of measuring the amount of soluble phosphate, try ICP-OES for Phosphorus. This method requires preparation of standards and controls as well as addition of internal standard to samples for quality assurance. The run can be automated and you can have the results the next morning if you put the run overnight. Over 30 samples can be done in an hour - that is what I call efficient. Try it.
Various methods are available for determination of soluble phosphate in waste water. Spectrophotometry, ion -chromatography and ICP_OES. Use suitable SRM for calibration.
Hello Kamal,
In our lab we use Hach kits to measure Phosphate, perhaps this might be an option for you?
Best wishes,
Ciara
Sample is first hydrolysed with acid and digested (heated) which breaks down strong chemical bonds and allows all phosphorus in the sample to be measured colorimetrically.
Hello dear friends I need help regarding Phosphate solubilizing activity because I,m working on endophytic bacteria,s so I want to determin this observation from bacteria,s if anyone know about this observation, so kindly send to me protocol of this observation as well as other plant growth promoting observation protocols Thanks
Phosphorus determination in biological fluids
The use of detergent to solubilize proteins in the determination of serum phosphorus was reported by several researchers, but every modification for the ingredients in the final reagent involve a change of the used detergent. In past years, I tried Teepol 610 S, an anionic detergent, which prevented the acid precipitation of serum protein while allowing the formation of heteropoly blue, the term given to the mixture of reduced phosphomolybdic acids measured colorimetrically for the quantitation of phosphorus through an MTP procedure.
Reagents
Sol. A: dissolve in 25 ml of dH2O, 2.2 g ammonium molybdate, then add 18 ml of conc. H2SO4 and the mixture is cooled to room temperature.
Sol. B: dissolve in 25 ml dH2O, 4.0 g ferrous ammonium sulfate (FAS), followed by the addition of 10 ml of conc. H2SO4 and the solution is cooled to room temperature.
Teepol 10%: dilute 10 ml of Teepol 610 S stock solution (Fluka cod. 86350) to 50 ml with dH2O, with gentle heating not over 56 °C followed by the addition of sufficient water to bring the volume to 100 ml.
One Reagent: mix Sol. A and Sol. B and add 300 ml of dH2O, then 40 ml of Teepol 10%, dissolve 0.25 g of polyvinylpyrrolidone (PVP in the range from 10K to 60K m.w.), and brought to a volume of 1000 ml by the addition of deionized water, and store at room temperature resulting stable 1 year.
Phosphorus Stock Standard 1 mg/ml: weigh 439.4 mg of dried and desiccated potassium dihydrogen phosphate and dissolve in dH2O to a 100 ml volume, and then add 5 ml chloroform as preservative. Store at 4 °C. Phosphorus Working Standard 100 µg/ml: dilute 10 ml of the stock standard to 100 ml with dH2O.
Procedure
Dispense in wells 25 µl of serum or and add 150 µl of one reagent ad incubate for 6 min. and then read the Abs at 650 nm. and calculating against standard and zero instrument with bank.
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