What is the method of preparation of phosphate buffer at pH 4.7 for alkaloid determination ?

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W.J. Colonna

Dear Y K D D Thathsarani,

I can't explain why the dibasic sodium phosphate did not dissolve in water. Its solubility is ~118 g/L.

Attached below is a recipe for citrate-phosphate buffer.

I hope this information helps you.

Bill Colonna

Dept. Food Science & Human Nutrition, Iowa State University, Ames, Iowa, USA [email protected]

Randolph Arroo

The anhydrous salt will dissolve eventually, but you may want to use the dihydrate salt which dissolves much quicker.

Phil Käding

Assessing Water Quality: Ensure that the water used is of high purity. Impurities in water can significantly affect solubility and buffer pH. Deionized or distilled water is recommended.

Temperature Adjustment: Increasing the temperature of the solvent (water) can enhance the solubility of sodium phosphate dibasic dihydrate. Heating the water to 30-40°C before adding the salt may facilitate dissolution without decomposing the salt or affecting its buffering capacity.

Incremental Addition: Gradually add sodium phosphate dibasic dihydrate to the water while continuously stirring. This method prevents the formation of aggregates that are difficult to dissolve.

Use of a Magnetic Stirrer: Employ a magnetic stirrer for uniform mixing. Continuous stirring for an extended period may be necessary until complete dissolution is achieved.

Adjusting to Desired pH with Citric Acid

Preparation of Citric Acid Solution: Dissolve citric acid in deionized or distilled water to create a concentrated solution.

Gradual Addition to Phosphate Solution: Add citric acid solution dropwise to the sodium phosphate solution under continuous stirring. Monitor the pH closely using a calibrated pH meter.

pH Adjustment: Continue the addition until the pH reaches 4.7. The precise amount of citric acid needed can vary; thus, incremental addition and continuous pH monitoring are crucial.

Troubleshooting

If solubility issues persist, consider using a sonication bath to facilitate dissolution. Ultrasonic waves can help break down aggregates, enhancing solubility.
Verify the purity and quality of the sodium phosphate dibasic dihydrate. Impurities can significantly impact solubility and buffer efficacy.
Reassess the concentration of sodium phosphate dibasic dihydrate. Overconcentration may exceed the solubility limit under the given conditions.