Development and validation of analytical method for quantification of pfáffico acid in Hebanthe eriantha

Author(s)

Kelly de Paula Souza

• Document type: Master's Dissertation
• Press: Campinas, SP.
• Institution: Universidade Estadual de Campinas (UNICAMP). Instituto de Química
• Defense date: 2011-02-17

Examining board members

Susanne Rath; Fernando Batista Da Costa; Ivo Milton Raimundo Júnior

• Advisor: Susanne Rath; Marili Villa Nova Rodrigues

Abstract

Hebanthe erianthe (Amaranthaceae) is known as "Brazilian ginseng¿, due to the similarity of its roots with those of Panax ginseng (Araliaceae), the "Korean ginseng¿. Its roots, with proved antineoplastic activity, are constituted by saponins, which during hydrolysis, release saccharides and free molecules of pfaffic acid, a nortriterpene. Pfaffic acid is an excellent option for phytochemical marker of H. erianthe, because it distinguishes it from Pfaffia glomerata, often employed as adulterant. The aim of this work was the development and validation of an analytical method for determination of pfaffic acid in H. erianthe. The development of sample preparation included the optimization of the extraction and hydrolysis steps of the saponins root, through univariate analysis and experimental design (central composite), respectively. The sample preparation consisted of extraction of saponins with 80% (v/v) ethanol, in two steps of 2 h, followed by hydrolysis with 1.8 mol L HCl during 4 h, at 120 °C. The pfaffic acid was quantified by high performance liquid chromatography, employing a C18 column, with isocratic elution and mobile phase constituted by a mixture of 0.1% (v/v) formic acid solution and methanol (18:82, v/v). The detection was performed at 205 nm. The identity confirmation of the analyte was carried out using liquid chromatography coupled to mass spectrometry, with atmospheric pressure chemical ionization. The method was validated and the following parameters were established: linearity (0.9999), linear range (0.04% to 12.8% m/m), intra-day precision (4.8%, n=7) and inter-day precision (5.2%, n=10), detection limit (0.01% m/m) and quantification limit (0.04% m/m). The accuracy was evaluated by comparing two methods, validated in two different laboratories. The validated method was applied for the determination of pfaffic acid in four commercial samples of roots (AU)

• FAPESP's process: 09/06738-6 - Development and validation of analytical method for quantitation of pfaffic acid in Pfaffia paniculata
• Grantee: Kelly de Paula Souza
• Support Opportunities: Scholarships in Brazil - Master