Purification of clavulanic acid using zeolites

Author(s)

Marcus Bruno Soares Forte

• Document type: Master's Dissertation
• Press: Campinas, SP.
• Institution: Universidade Estadual de Campinas (UNICAMP). Faculdade de Engenharia de Alimentos
• Defense date: 2008-04-25

Examining board members

Maria Isabel Rodrigues; Fernando Antonio Cabral; Marlei Barboza Pasotto; Daniel Ibraim Pires Atala

• Advisor: Maria Isabel Rodrigues; Francisco Maugeri Filho

Abstract

In this research work, a method for separation and purification of clavulanic acid from a fermented medium was studied using natural and synthetic zeolites. The natural zeolite (ZN) and synthetic faujasite (13X) were modified by ionic exchange with different compensation cations (Na+1, K+1, Ca+2, Ba+2, Mg+2, Sr+2). The kinetic analysis by adsorption of clavulanic acid indicated that the zeolite 13X-Na was the most promising for the adsorption of the related compound. At the equilibrium conditions, the retention was about 17.4% (CAC*/CAC0 = 0.826), where the amount adsorbed clavulanic acid relative to the amount of zeolite (q*) was 0.4927 mg/g. The average diameter of a clavulanic acid molecule (DAC) was estimated as 9.6 A. The composition of the zeolite 13X-Na was characterized in terms of the ratio Si/Al (1.5), density (dz = 2.248 g/cm3), superficial area (SBET = 444.86 m2/g), total pore volume (Vpores = 0.308 cm3/g), micropore volume (Vmicro = 0.203 cm3/g) and average pore diameter (Dpores = 28 A). Accordingly, particle and bed porosity (ep and eb) were calculated as 0.69 and 0.85 respectively. Solutions of pure clavulanic acid were obtained in semi-preparative scale HPLC. The adsorption isotherm for 13X-Na and solutions of pure clavulanic acid were determined for the temperatures of 10, 15 and 200C, showing at increasing temperature, the performance of the process decreases. Addition of fermented medium in the fixed bed column of Zeolites 13X-Na improved the separation process between clavulanic acid and contaminant proteins, using as effluent phosphate buffer (0.2 M and pH 6.2) and zeolites with particle diameters in the range of 0.053 to 0.062 mm. The separation efficiency (ES), factor concentration (FC), purification factor, referred to proteins (FPP), and purification factor referred to the contaminants (FPC) were determined, as ES = 0.38, FC = 0.40, FPP = 1.25 and FPC = 1.28 respectively (AU)