Detection of D-Dopa in levodopa tablets by derivatization coupled with LC-MS/MS.

Chiral impurity analysis presents significant challenges due to limitations of detection methods. Levodopa (L-Dopa) is the primary medication for Parkinson's disease treatment for over 50 years. However, the detection of its chiral impurity is difficult due to the similarity of polarity. This study presents a novel analytical method for the quantitative detection of trace amount of d-Dopa, as well as other three impurities. The method demonstrated excellent chromatographic resolution for d-Dopa and l-Dopa (resolution = 4.15) and achieved a derivatization efficiency of 97 % using a chlorine-labeled chiral probe (D-BPCl) for chemical derivatization, followed by analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) on a conventional C18 column. Meanwhile, d-BPCl showed strong chiral selectivity on d-Dopa, and the ratio of mass spectrometric response for labeled d-Dopa to that of l-enantiomers was 2.78-fold under the same condition. The developed method demonstrates high sensitivity with detection limits of 7.88 μg l for d-Dopa, and 9.85 μg l for l-Dopa, with good linearity for every analyte (R > 0.99), recoveries ranging from 94.1 % to 106.8 %, and high repeatability with relative standard deviation (RSD) values predominantly below 5 %. Then the method was applied to analyze fifteen batches of levodopa tablets from five manufacturers. d-Dopa was successfully detected with the corresponding content ranging from 0.15 % to 0.44 %, which suggests that d-Dopa are more likely generated during the production process. The study underscores the significant safety concerns regarding Dopa-containing medications.