Tropical Journal of Pharmaceutical Research

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Liquid Chromatography – Mass Spectrometry Method for the Simultaneous Determination and Confirmation of Seven Active Components in Chinese Medicine Kumu Injection

Z Lai, H Liao, S Ip, Y Yi, S Shi, J Su, X Lai, Z Su, Z Lin


Purpose: To develop and validate a simple and selective high  performance liquid chromatography photo diode array mass spectrometry (HPLC-PDA-MS/MS) method for simultaneous determination and confirmation of seven major active alkaloids (6-Hydroxy-ß-Carboline-1-carboxylic acid, ß-Carboline-1- carboxylic acid, ß-Carboline-1-propanoic acid, 3-Methylcanthin-5,6-dione, 4-Methoxy-3-methylcanthine- 5,6-dione, 5-Hydroxy-4-methoxycanthin-6-one, 4,5-Dimethoxycanthin-6-one) in  Kumu injections (KMIs)
Methods: For the analysis of the preparation, the optimal chromatographic condition was achieved on a Phenomenex Gemini C18 column with  gradient elution of 25 mM aqueous ammonium acetate (pH = 4.0 adjusted by glacial acetate acid) and acetonitrile with flow rate at 1.0 mL/min,  column temperature at 35oC and detection wavelengths at 245, 260 and 271 nm.
Results: Excellent linear behavior over the investigated concentration ranges was observed with regression coeffcient (R2) > 0.9997 for all analytes. Intra- and inter-day precisions for all studied constituents ranged from 0.20 to 1.80 %. Recoveries of the assayed constituents were in the range of 98.73 to 100.34 %. The results showed the contents of these seven marker compounds differed significantly among different batches of KMIs both from the same and different manufacturers.
Conclusion: The validated method was reliable, accurate, repeatable and can be applied to routine quality assessment of these active components in KMIs.

Keywords: Alkaloids, High performance liquid chromatography, Photo diode array, Mass spectrometry, Kumu injection, Quality control
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