الاثنين، 18 أبريل 2022

Phosphorus

 Phosphorus method

 Test solution

 Use a volumetric flask with a suitable volume for preparation of a  solution containing about 5 mg per millilitre of dry polysaccharide.

 Transfer the  contents of a container quantitatively to the flask and dilute to volume with water R.

  Dilute the solution so that the volume used in the test (1 mL) contains about 6 µg of  phosphorus. 

Transfer 1.0 mL of the solution to a 10 mL ignition tube.   

 Reference solutions

 Dissolve 0.2194 g of potassium dihydrogen phosphate R in  500 mL of water R to give a solution containing the equivalent of 0.1 mg of  phosphorus per millilitre. 

Dilute 5.0 mL of the solution to 100.0 mL with water R.  Introduce 0.5 mL, 1.0 mL and 2.0 mL of the dilute solution into 3 ignition tubes.

 Prepare a blank solution using 2.0 mL of water R in an ignition tube.

 To all the tubes add 0.2 mL of sulfuric acid R and heat in an oil bath at 120 °C for 1 h  and then at 160 °C until white fumes appear (about 1 h). Add 0.1 mL of perchloric  acid R and heat at 160 °C until the solution is decolorised (about 90 min). 

Cool and  add to each tube 4 mL of water R and 4 mL of ammonium molybdate reagent R.  

Heat in a water-bath at 37 °C for 90 min and cool.

 Adjust the volume to 10.0 mL with  water R. The blue colour is stable for several hours.

Measurements

Measure the absorbance of each solution at 760 nm using the blank as the  compensation liquid. 

Draw a calibration curve from the absorbances of the 6  reference solutions and the corresponding protein contents and read from the curve  the content of protein in the test solution.

References

Ph. Eur. method 2.5.18


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