Determination of huperzine A in formulated products by reversed-phase-liquid chromatography using diode array and electrospray ionization mass spectrometric detection

• Published in: Phytomedicine (Stuttgart) 2003-03, Vol.10 (2), p.200-205
• Main Authors: Yang, Q.P., Kou, X.L., Fugal, K.B., McLaughlin, J.L.
• Format: Article
• Language: English
• Subjects: Alkaloids; Biological Products - chemistry; Chromatography, High Pressure Liquid - methods; Club-mosses; Clubmosses; Health aspects; HPLC-UV; HPLC/MS; Huperzia; Huperzia serrata; huperzine A; Liquid chromatography; Mass spectrometry; Medicinal plants; Medicine, Botanic; Medicine, Herbal; Physiological aspects; Plant Extracts - chemistry; Plant Extracts - isolation & purification; Sesquiterpenes - chemistry; Sesquiterpenes - isolation & purification; Spectrometry, Mass, Electrospray Ionization - methods; Spectrum Analysis
• ISSN: 0944-7113; 1618-095X
• DOI: 10.1078/094471103321659942

A precise and selective high-performance liquid chromatographic (HPLC) method with diode-array detection for quantifying huperzine A in formulated products was developed and validated. A liquid chromatographic-mass spectrometric (LC/MS) procedure was devised to confirm the HPLC method. Huperzine A was dissolved in 1,2-dichloroethane, chromatographed on a YMCBasic C18 column, and detected at 308 nm. A gradient mobile phase of 10 mM ammonium acetate (pH = 3.5) – methanol was used. Identification was based on retention time, UV spectra and mass spectra by comparison with a commercial standard. The UV peak areas were used for quantitation of huperzine A content. The correlation coefficient (R 2) of the calibration curve was 1 over the range 0.8–11.6 μg/ml. Overall recovery of huperzine A was 103.9% ± 1.8 (mean ± SD). Relative standard deviations for intra- and interday precision were