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-1 . 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-1 for levodopa impurity B; 5.26 and 15.9 ng mL-1 for levodopa impurity C; 0.833 and 2.53 ng mL-1 for methyldopa; 3.31 and 10.0 ng mL-1 for methylcarbidopa; 1.67 and 5.06 ng mL-1 for entacapone impurity C; and 0.61 and 1.86 ng mL-1 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.

Determination of Zoledronic Acid and Its Related Substances by High Performance Liquid Chromatography with Evaporative Light Scattering Detection.

A method has been developed and validated for analysis of zoledronic acid (ZOL) and its related substances by ion-pair reversed-phase high performance liquid chromatography with evaporative light scattering detection (ELSD). Chromatographic separation was achieved with gradient elution by using a C18 column, mobile phase containing 12 mM ammonium acetate buffer and 35 mM n-pentylamine, whose pH value is 7.0, and 5% acetonitrile. The mobile-phase flow rate was 1.0 mL/min. The calibration plot was linear in the range from 0.4 mg/mL to 6.0 mg/mL for ZOL and from 6.25 µg/mL to 100 µg/mL for its related substances. ZOL and its related substances, namely imidazole-1-yl-acetic acid, phosphate, phosphite and degradation products did not interfere with each other. The method was rapid, linear, accurate and reproducible. The high performance liquid chromatographic method that has been developed to determine the related substances and assay of ZOL can be used simultaneously to evaluate the quality of regular samples. It can be also used to test the stability samples of ZOL.