A rapid liquid chromatography/tandem mass spectrometry method for simultaneous determination of levodopa, carbidopa, entacapone and their six related compounds in film-coated tablets.

RATIONALE

A liquid chromatography/tandem mass spectrometry (LC/MS/MS) method has been developed and validated to determine levodopa, carbidopa, entacapone, and corresponding six related substances - levodopa impurity B, levodopa impurity C, methyldopa, methylcarbidopa, entacapone impurity C, and entacapone impurity A - in film-coated tablets for the first time.

METHODS

Chromatographic separation was achieved with a gradient elution by using a C18 column, a mobile phase containing 0.5% formic acid in water and 0.5% formic acid in methanol. The mobile phase flow rate was 0.5 mL min . The UV detector was set at 280 nm and the triple quadrupole mass spectrometer was used in multiple reaction monitoring (MRM) mode.

RESULTS

The limit of detection (LOD) and limit of quantification (LOQ) results were 1.3 and 3.94 ng mL for levodopa impurity B; 5.26 and 15.9 ng mL for levodopa impurity C; 0.833 and 2.53 ng mL for methyldopa; 3.31 and 10.0 ng mL for methylcarbidopa; 1.67 and 5.06 ng mL for entacapone impurity C; and 0.61 and 1.86 ng mL for entacapone impurity A.

CONCLUSIONS

The method was rapid, linear, accurate, and reproducible. The LC/MS/MS method that was developed to determine the related substances and assay of levodopa, carbidopa, and entacapone can be used to evaluate the quality of regular samples in the pharmaceutical industry. It can be also used to test the stability of samples.