Hey Shashikant,

Be aware that C (in C1V1=C2V2) stands for molar concentration (mol/l), not ratio (molar or otherwise). In the paper you are using, the authors say that you need 5% formic acid. Although they did not say it specifically, I asume what they are referring to is a volume ratio. This being said, if you need to make a mobile phase manually, you can do it by mixing 47.5 parts of water, 47.5 parts of ACN and 5 parts of formic acid (if you need one liter of solution, it would be 475 ml H20, 475 ml ACN and 50 ml HCOOH).

Good luck!

Shashikant, Polina is correct. You stated that the authors wrote that the solution was a volume/volume solution, so use a graduated cylinder(s) to prepare the mixture.

Dear Shashikant Kotwal

Please do not think it complex.

Just follow this formula C1V1 = C2V2

Question: Example for 2000 mL mobile phase solution of acetonitrile and water (1:1).

Ans: 950 mL Acetronitrile + 950 mL Water + 100 mL formic acid = 2000 mL

Thank you

good question

and it is already answered by Polina D. Blagojevic and William Letter

the 5% is mostly referring to the volume ratio, first, you can prepare acetonitrile:water solution by mixing equal amounts of both solvents, say 50 ml and 50 ml.

then to 95 ml of the acetonitrile:water solution you add 5 ml of formic acid to make volume 100 ml, and like this, you get "5% formic acid in acetonitrile/water 1:1 v:v "

Thank you all for the help! Separation did not go so well, but will keep trying. :)