Development and Validation of a Simple HPLC-UV Method for Analysis of Harmine

Authors : Galip Mesut Demir, Afife Büşra Uğur Kaplan, Mine Gülaboğlu, Meltem Çetin

Pages : 1096-1102

View : 58 | Download : 60

Publication Date : 2025-07-15

Article Type : Research Paper

Abstract :In this study, a rapid, simple, and sensitive HPLC-UV method was developed and validated in accordance with ICH Q2 (R1) guideline for the quantitative analysis of harmine (HRM), a main β-carboline alkaloid found in the Peganum harmala (Uzerlik) plant. The method was established based on the UV spectral analysis, which indicated maximum absorbance for HRM at 243 nm. A mixture of methanol and ultrapure water containing 1% formic acid (50:50, v/v) was selected as the mobile phase. In the separation carried out under isocratic conditions, the retention time of HRM was determined as 5.5 min, and the entire analysis was completed in under 8 min. The method demonstrated excellent linearity within the concentration range of 1–36 µg/mL, with a correlation coefficient (R²) of 0.99998. The accuracy and precision parameters of the method were found to be within acceptable limits, with relative error (RE) and relative standard deviation (RSD%) both below 2. The limits of detection and quantification (LOD and LOQ) were 0.269 µg/mL and 0.816 µg/mL, respectively. Also, it was determined that HRM standard solutions were stable for up to 7 days when stored at different temperatures (−20°C, 2–8°C, 25°C). In conclusion, this method offers a reliable alternative for the quantitative determination of HRM with high analytical quality, reproducibility, and short time. It can be widely used in routine laboratory practices for the monitoring of HRM in pure form and in various pharmaceutical formulations.
Keywords : Harmine, HPLC-UV method, Peganum harmala, Stability, Validation

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