Chromatography Forum

Has anybody here had experience running Formic Acid in Povidone using either USP or EP monographs? I am having an issue getting my retention time down to 11 minutes at 30 C with the column I currently have (Hamilton HC-75 H+ 4.1 x 250mm). If I bump the temperature up to 60 C I get a retention time of 10.9 min, but as far as I can tell, I only have +/- 10% on the temperature to keep compliant with USP methods. Any suggestions on a better column, the USP calls for a L17 column? The Hamilton can only handle 400 psi, which is my biggest constraint. Below is the EP monograph, which is the same as the USP. Any help would be appreciated.

Formic acid. Liquid chromatography (2.2.29).
Test solution. Dissolve a quantity of the substance to be
examined equivalent to 2.0 g of the anhydrous substance in
water R and dilute to 100.0 mL with the same solvent (test
stock solution). Transfer a suspension of strongly acidic
ion-exchange resin R for column chromatography in water R
to a column of about 0.8 cm in internal diameter to give a
packing of about 20 mm in length and keep the strongly acidic
ion-exchange resin layer constantly immersed in water R. Pour
5 mL of water R and adjust the flow rate so that the water
drops at a rate of about 20 drops per minute. When the level
of the water comes down to near the top of the strongly acidic
ion-exchange resin layer, put the test stock solution into the column.
After dropping 2 mL of the solution, collect 1.5 mL of
the solution and use this solution as the test solution.
Reference solution. Dissolve 0.100 g of anhydrous formic
acid R in water R and dilute to 100.0mL with the same solvent.
Dilute 1.0 mL of the solution to 100.0 mL with water R.
Column:
— size : l = 0.25-0.30 m, Ø = 4-8 mm;
— stationary phase: strongly acidic ion-exchange resin R for
column chromatography (5-10 μm);
— temperature: 30 °C.
Mobile phase: dilute 5 mL of perchloric acid R to 1000 mL
with water R.
Flow rate: adjusted so that the retention time of formic acid is
about 11 min.
Detection: spectrophotometer at 210 nm.
Injection: 50 μL.
System suitability : reference solution:
— repeatability : maximum relative standard deviation of
2.0 per cent after 6 injections.
Limit:
— formic acid : not more than 10 times the area of the principal
peak in the chromatogram obtained with the reference
solution (0.5 per cent).

Here is the exact columns used in Ph. Eu.

Column 0.8 cm ID: Michel-Miller chromatography column from Aldrich - resin: IR 120 from Rohm and Haas Column Shim-pack SCR-101H (30 cm x 7.9 mm ID): SHIMADZU CORPORATION or equivalent

And you can use the link below to search the columns used in Ph. Eu.

https://extranet.edqm.eu/page7/page2/page2.html

I have done the formic acid analysis in povidone with same column i.e (Hamilton HC-75 H+ 4.1 x 250mm)and I am getting the retention time at 11.0 mins at 30 C.My issue is that I am getting unwanted peaks till 350 mins.Please suggest me a solution for this issue.This situation is urgent kindly do respond as earliest.

Procedure as per EP monograph

Formic acid. Liquid chromatography (2.2.29).
Test solution. Dissolve a quantity of the substance to be
examined equivalent to 2.0 g of the anhydrous substance in
water R and dilute to 100.0 mL with the same solvent (test
stock solution). Transfer a suspension of strongly acidic
ion-exchange resin R for column chromatography in water R
to a column of about 0.8 cm in internal diameter to give a
packing of about 20 mm in length and keep the strongly acidic
ion-exchange resin layer constantly immersed in water R. Pour
5 mL of water R and adjust the flow rate so that the water
drops at a rate of about 20 drops per minute. When the level
of the water comes down to near the top of the strongly acidic
ion-exchange resin layer, put the test stock solution into the column.
After dropping 2 mL of the solution, collect 1.5 mL of
the solution and use this solution as the test solution.
Reference solution. Dissolve 0.100 g of anhydrous formic
acid R in water R and dilute to 100.0mL with the same solvent.
Dilute 1.0 mL of the solution to 100.0 mL with water R.
Column:
— size : l = 0.25-0.30 m, Ø = 4-8 mm;
— stationary phase: strongly acidic ion-exchange resin R for
column chromatography (5-10 μm);
— temperature: 30 °C.
Mobile phase: dilute 5 mL of perchloric acid R to 1000 mL
with water R.
Flow rate: adjusted so that the retention time of formic acid is
about 11 min.
Detection: spectrophotometer at 210 nm.
Injection: 50 μL.
System suitability : reference solution:
— repeatability : maximum relative standard deviation of
2.0 per cent after 6 injections.

regards
Deepti

We're running this analysis on a regular basis. We've solved the problem by first packing a pasteur pipette with a strongly acidic ion-exchange resin, transferring the eluate to a HPLC-vial and analysing it on a Benson BP-OA column(300x7.8 mm) Flow: 0.75ml/min. Temp: 30 C.