Skip to main content

Chromatography

               An analytical technique which is used for the isolation, identification and separation of compounds based on differences in affinity for a stationary phase and a mobile phase. In chromatography, the stationary phase may be a solid, liquid, gel or a solid/liquid mixture.. and mobile phase may be a liquid or a gas. 


Principles of Chromatography

"A mixture of compound is allowed to pass through two phases called stationary phase and mobile phases, the separation of compounds can be seen according to their Resolution Front (Rf) values. " Basically it is adsorption and partition. 
The Resolution front (Rf) value is defined as the ration of distance traveled by the sample solute divided by distance traveled by the solvent front. 
During chromatographic separation, there are two interactions occur..
1. Adsorption: separation can be achieved by the differential adsorption of substances by the mobile and stationary phases. 
2. Partition: when a solute is dissolved in two immiscible liquids of equal volume, the ration of its concentration in two phases at a given temperature and equilibrium is called partition coefficient. 

Types of Chromatography
(Click on the type of chromatography to know the details)

6) Gas-Liquid Chromatography (GLC)
7) High Performance Liquid Chromatography (HPLC) 
Labels:

Comments

Post a Comment

Popular posts from this blog

Statistics in Flow Cytometry Data and "MFI" values

                                 The speed of the flow cytometry offers wide range of data points and data plots. Due to its sensitivity and versatility, it has been used extensively. Flow cytometer can detect up to 1000s of cells per second.. But, it should be noted that the flow cytometry also involves the statistics, its significance, calculating the fluorescence intensity..                Statistics in flow cytometry involves, total number of data points acquired, percentage population, most importantly mean and median fluorescence intensity, and others. Most of us get confused about the term "MFI", whether it means mean fluorescence intensity or median fluorescence intensity.. Truth is, both.. It means it can be described as either mean or median fluorescence intensity. Some researchers use Mean for MFI and some use Median for MFI values.  (Keep...
1 comment
....CONTINUE READING

Protocol: Sample preparation for Cell Cycle Analysis by Flow Cytometry

Protocol:  Preparation of Samples for Cell Cycle Analysis by Flow Cytometry: 1) Suspend approximately million cells (10^6) in 1mL of phosphate-buffered saline solution (1x). And with the help of pipette, gently aspirate about 7-8 times, to make the single cell suspension.  2) Take a centrifuge tube and add 4mL of 70% ethanol. (70% ethanol should be one ice before using). and add 1mL of cells in PBS to this tube. Keep the tubes on ice for minimum 2 hours at 4℃.. This step will fix the cells.  ( cells can kept at 4 ℃ for a week).  3) Centrifuge the ethanol fixed cells for 5 minutes at 300g. Carefully take out the supernatant. Do not disturb the pellet.  4) Add 5mL of cold 1x PBS to this pellet and suspend the cells using pipette.  5) Centrifuge the tubes at 300g for 5 minutes. and discard the supernatant. Again do not disturb the pellet.  6) if required, step 5 can be repeated for one more time.  7) Add 1mL of Propidium Iodide (PI...
Post a Comment
....CONTINUE READING

Incubators

            In biological sciences or bio medical sciences, the incubators are the devices which are used to maintain or grow microorganisms or cell cultures. Main task of an incubator is to maintain the optimal temperature, humidity, carbon monoxide, or CO2 and O2 content.  Integrated Scientific Solutions     Incubators are regularly, in fact daily are used in clinical or research areas such microbiology, molecular biology, cell biology, etc. Commonly they are used to culture or grow bacteria as well as eukaryotic cells. Incubators are insulated boxes in which the required temperature can be adjusted depending on the cells or bacteria to be grown or cultured. The temperature can be up to 65℃ to  70 ℃.. Most commonly used incubators are 37 ℃ and 30 ℃ incubators. 37 ℃ are used for growing bacteria such as E.coli bacteria. and 30 ℃ are commonly used for growing yeast cells. There are special incubators which maintain the humidity...
Post a Comment
....CONTINUE READING