Methods Column chromatography was set up using a 60 ml syringe clamped to a ring stand. The syringe was plugged using a small amount of cotton. An Erlenmeyer flask was placed underneath the column to collect any excess material. Eleven 20 ml vials were labeled numbers 1 through 11. Silica gel was measured to 42 ml using a graduated cylinder and then moved into a 150 ml beaker. A slurry was created by adding 50 ml of hexane to the beaker containing the silica gel. The slurry was stirred for about one minute. This was the stationary phase for the column chromatography. Pour the slurry into the column that is plugged with cotton. Hexane was added to the beaker to move all the slurry into the column. More hexane was added to the column until the…
Introduction Separation, a process by which the distinguishing between substances through physical or chemical means in a mixture may be carried out. A widely employed technique for this purpose is called chromatography. 1 There are many different types of chromatography, including gas-liquid, paper, column and so on. 1 Chromatography differentiates by allowing the differences in polarity, vapor pressure, solubility and so on of different substances to be observed. 1 In our endeavor, we will be…
ABSTRACT: In this experiment, we determined the relative polarity of substances using the principles of chromatography. Three substances were tested using a thin layer chromatography. The column chromatography tested a mixture of two substance which were separated as a result of the chromatography. The results for both the thin layer and column chromatography supported the hypothesis. Therefore, chromatography is an effective tool that can be used to determine relative polarities. INTRODUCTION:…
Initially, the protein extract solution was subject to ion exchange column chromatography using SP Sepharose Fast Flow (GE Healthcare, Sweden). The column (with 5 ml of stationary phase) was equilibrated with five column volumes of sodium acetate/acetic acid 10mM pH5 and 10 ml of protein extract containing bacteriocin were injected into this. After washing the column with sodium acetate/acetic acid 10 mM pH5 (Buffer A), elution was performed using a linear gradient from sodium acetate/acetic…