Chromatography

potential designer modification of banned drugs and show the same elemental composition and yielding identical nominal, exact masses and general analytical properties which make the potential of misidentification increase. Electron- ionization gas chromatography- mass spectrometry (EI-GC-MS) technique was used as initial methods to differentiate and separate the target compounds. In addition, consecutive confirmation methods was provided by…

MDMA (3,4-methylenedioxymethamphetamine), commonly referred to as ecstasy, and methamphetamine are central nervous system stimulants, which can produce euphoria, enhanced mental and emotional clarity, sensations of lightness and floating, and other hallucinations. (Chandra et al. 2015) Due to their effects, MDMA and methamphetamine are used in Drug-Facilitated Crimes (DFC), intended to impair behaviour, perceptions, or decision-making capacity to take advantage of an impaired person without…

grown in medium containing sorbital and betaine. Ferritin synthesis was induced in mid-log cells using isothiopropyl thiogalactoside (IPTG), and the soluble ferritin was isolated, after sonication of the bacterial cells, by fast protein liquid chromatography (FPLC) on a Fast Q-Sepharose matrix. Protein purified to homogeneity was sterilized by filtration through a 0.45 µm membrane and stored at 4 °C for less than 1 week before mixing with…

Transformation of pGlO plasmid to an E.coli bacteria cell We pipetted 250 ul of CaCl2 transformation solution to into two separate tubes. Next, we used a sterile loop to transfer two to four larger bacteria colonies from an E.coli plate into each of the two tubes. The bacteria was observed under UV light to make initial observations. We pipetted 10 ul of pGLO plasmid (0.08 ug/ul) into one of the tubes. Both tubes were incubated on ice for ten minutes. After ten minutes of incubation, the tubes…

Anion exchange chromatography: In anion exchange chromatography negatively charged ions will bind to the column. This will leave positively charged ions flushed through the column. By picking a pH of 6.5, protein X will flush through the column while protein Y and Z will stick to the sides of the column. Then by changing the pH to 8 it would make protein Z negatively charged and will be stuck in the column as protein Y is flushed to the bottom of the column. Cation exchange chromatography: This…

4.3 High Performance Liquid Chromatography (HPLC) An HPLC analysis was carried out for the Fraction C2-7 for confirmation of the results obtained by TLC analysis. According to the HPLC chromatogram, there is only one peak was observed at the retention time of 4.510 minutes. Therefore, it can be concluded that there is only one compound presents in the Fraction C2-7. Basically, the first few small peaks in the chromatogram which is around 2-3 minutes are ‘noise’ from the injection that been made…

Introduction: LC-MS is used for the bioanalysis of small molecules drugs, which has molecular mass of less than 800 Da. Now a days, it is also frequently used for the bioanalytical determination of biopharmaceuticals.1 In pharmaceutical R & D, it takes a year to develop a new marketing drug identified from thousands of new compounds. The drug discovery is divided into early and late phase, during which pharmacokinetics study or bioanalysis of a drug is done. Once, the bioanalysis of a drug is…

During this experiment that was conducted in partners, the concentration of RNase H that was purified in the previous experiment #6 was determined and also the concentration of an unknown given solution using their absorbance values obtained from the spectroscopy apparatus. These concentrations were calculated using the equation of the line produce with the values of the concentration (x-axis) and corresponding absorbance (y-axis) of five protein bovine gamma globulin standard solutions. In…

1. Abstract In this study, reverse micellar extraction of asparaginase model system was carried out using CTAB/Iso-octane/hexanol/butanol system to optimize the forward and back extraction efficiency. By using Taguchi’s orthogonal array (OA), maximum forward extraction efficiency of 55.0%, 61.0% and 54% was achieved with an aqueous phase pH of 11.0, 150mM CTAB/Iso-octane and 0.1 M NaCl, respectively. Similarly, Taguchi’s orthogonal array (OA) was applied to optimize the pH of stripping phase,…

successful separation of the pigments from the spinach leaves, the pigments are put into silica gel and the solvent is allowed to evaporate off so that the pigments (now in powder form) can be put through column chromatography to separate the chlorophyll and β-carotene pigments. Column chromatography is a means to divide chemical compounds from a mixture. Separation is achieved by eluting solvents through the column which hold similar properties to one or the other pigment. Chlorophyll, being a…