Method of analysis for determination of Curcumin and its impurities by HPLC

Buffer solution: 10 ml acetic acid in 500 ml water.

Mobile phase: Buffer solution: acetonitrile (50:50)

Chromatographic conditions:

Column : Alpha C18, 250 mm x 4.6 mm , 5 µ

Flow rate : 1.2 ml/min
Column oven temperature : 35⁰C
Wavelength : 425 nm
Injection volume : 20 µl
Needle wash : Water: Methanol (50:50)
Run time : 25 min
Diluent: Methanol

Sample preparation

Transfer 100 mg of powdered sample to 100 ml volumetric flask. Add about 30-40 ml of diluent.
Shake well and sonicate for 3 min. Make up to the mark with diluent. Shake well and filter through
0.45 µ syringe filter. Discard first few ml of the filtrate. Use the further filtered solution as sample
preparation.

Retention times:

Name Retention time

Bisdesmethoxycurcumin 11.7 min
Desmethoxycurcumin 13.0 min
Curcumin 15.0 min

Note: Proposed impurities are based on literature search. Individual RTs can be confirmed by
individually injecting the impurities.
Typical Chromatograms:

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Sample Preparation