Development and validation of a rapid RP-HPLC-DAD analysis method for the quantification of pilocarpine in Pilocarpus microphyllus (Rutaceae)

• Published in: Food and chemical toxicology Vol. 119; pp. 106 - 111
• Main Authors: Caldas Pereira, Rafael, Nonato, Carla de Fátima Alves, Camilo, Cicera Janaine, Melo Coutinho, Henrique Douglas, Rodrigues, Fabíola Fernandes Galvão, Xiao, Jianbo, da Costa, José Galberto Martins
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01-09-2018
• Subjects: Alkaloid; HPLC; Pilocarpine; Pilocarpus microphyllus; Quantification; Validation; Validation; Alkaloid; Pilocarpine; Quantification; Pilocarpus microphyllus; HPLC
• ISSN: 0278-6915; 1873-6351
• DOI: 10.1016/j.fct.2018.05.023

A method using high performance liquid chromatography with diode array detector (HPLC-DAD) for identification and quantification of pilocarpine in the extract of Pilocarpus microphyllus, popularly known as jaborandi. The analysis was conducted using RP-18 column (250 mm × 4.6 mm x 5 μm id) and a buffer solution composed of acidified water, phosphoric acid and triethylamine and methanol as a mobile phase in a flow rate of 1.0 mL/min and detection at 215  nm at 25 °C. Excellent linearity with r2 equal to 0.9999 was obtained. The recovery percentage was very satisfactory with values within the specifications. It is correct to affirm that the method has optimal intracurrent and intercurrent precision values with relative standard deviations of 0.1852% and 0.1932%, respectively. The robustness of the method, assessed through the Youden test, showed no significant influence of any of the evaluated parameters. In general, the method proved to be suitable for the intended purpose.
•The quantification of pilocarpine was validated by HPLC.•The analyzes confirmed above 97% purity of pilocarpine.•Linearity with r2 equal to 0.9999 was obtained.•The method has optimal intracurrent and intercurrent precision.