Journal of Chromatography & Separation Techniques
Open Access

Abstract

Gel Filtration Chromatography Technique as Tool of Simple Study Seminal Plasma Proteins in Domestic Animals

Vasconcelos André Belico de, Oliveira Jamil Silvano de and Lagares Monique de Albuquerque

Seminal fluid is the liquid component of sperm, providing a safe surrounding for spermatozoa. The seminal plasma has the feature common to many other body fluid, characterized by a high dynamic range of proteins, which visualizes physiological and biochemical processes of semen. As in other body fluids, it is convenient to distinguish in the seminal plasma between proteins and non-proteins. The molecular exclusion chromatographic technique presents important points to constitutional proteins preservation, in addition to the conventional phenomenon; witch exclusion hydrophobic interactions can provide a higher resolution in the chromatogram and do not provide nonspecific interactions between protein structures. The aim was the chromatography profile to simple study of proteins seminal plasma domestic animals (Stallions, Canine, and Goat), by the technique of gel filtration chromatography. The samples (seminal plasma proteins) were chromatographed in a Superose 12 HR 10/30, equilibrated with 25 mMTris-HCl (Sigma) with 0.15 M NaCl (Sigma), pH 7.4 at room temperature in a fast performance liquid chromatography (FPLC-system), using a flow rate of 0.5 mL.min-1. There was a molecular separation of significantly different molecular weights, which makes possible the logarithmic relationship between molecular weight and the elution volume. Calibration of the gel filtration column resulted in an equation y = ax + b, where the values of a and b were 5.43 and -2.175, respectively, and the corresponding errors were 0.115 and 0.256, respectively. The experimental error was less than 5% for most of the protein molecular masses. The work showed that the gel filtration chromatography technique provided an excellent analytical repeatability, and could therefore, be a valuable tool to the study preliminary of seminal plasma.