In a protocol book the composition of HPLC eluent is given as - "2mM sodium acetate in 0.75% (v/v) triethylamine:acetonitrile (97:3)". Could anyone suggest me how to prepare it step by step?
Typically these solutions don't need to be prepared with extreme precision. A simple protocol would be to add 656 mg of sodium acetate and 30 mL of triethylamine to a fresh 4L bottle of HPLC grade (or higher) acetonitrile. If you want to be more precise, you could measure out 3.970 L of acetonitrile, then add 30 mL of triethylamine and 656 mg sodium acetate. Either way, be sure to stir the solution thoroughly to dissolve all the sodium acetate.
Thank you William Sir,
But what exactly is the significance of the 97:3 ratio?
What I feel is:
We have to prepare 0.75% triethyl amine in water and mix it in acetonitrile in 97:3 ratio. Then add required amount of sodium acetate to get its final concentration as 2mM.
Am I right????
97 : 3 means that in a solution there must be 97 parts of the first constituent and 3 parts of the second, is important the order, for your formula, for example meaning - triethylamine: acetonitrile (97: 3) - if you prepare 100 ml you would have to add 97 ml of triethylamine and 3 ml of acetonitrile. But if the order is acetonitrile: triethylamine (97: 3) to prepare 100 ml you would have to put 97 ml of acetonitrile and 3 ml of triethylamine. All solvents must be HPLC grade. To prepare your mobile phase you must prepare a 2mM solution of sodium acetate diluted in deionized water. Subsequently you substitute a volume of 0.75% of the solvent solution triethylamine: acetonitrile. For example, if you prepared a liter of 2mM sodium acetate, take a volume of 7.5 ml of this solution and substitute it with 7.5 ml of your solvent preparation (97: 3).
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