Separation identification of amino acids - paper chromatography

the purpose of the experiment
. 1. Learning
Amino Acids
Basic Principles of Paper Layering
2. Mastering the Operation Techniques of Amino Acid Paper
(Experimental Principles
Paper Chromatography is the separation and identification of amino acid mixtures on
Biochemistry
Common techniques can be used for qualitative identification and quantitative determination of amino acid components of
protein
, and are also a separation analysis tool for qualitative or quantitative determination of
peptides
, nucleic acid bases, sugars,
organic
acids, vitamins, antibiotics and other substances. Paper lamination method is the distribution of paper as an inert support method, wherein the water adsorbed on filter paper cellulose is fixed phase, the organic solvent used for layering is the flow phase. In the lamination, the sample point is about 2 to 3 cm from one end of the filter paper, the point is called the origin, and then in a closed container the layering solvent along one direction of the filter paper, so that the mixed amino acids are constantly distributed in the two phases, due to different distribution coefficients (Kd), the results are distributed in different positions of the filter paper. The position of matter on the paper layer map after it is separated can be represented by a ratio shift value (rate of flow, Rf). The so-called RF refers to the ratio of the distance from the origin to the center of the amino acid dwell point (also known as the analysis point) (X) to the distance (Y) of the origin to the solvent frontier in paper analysis: the RF value of a substance is constant under certain conditions The size of the Rf value is related to the structure, properties, solvent system, temperature, humidity, type and quality of the layering filter paper.
equipment and
reagents
)
(i) equipment
1.layering cylinder
2. dot-like capillary tube
3.small
beech 4.
cultured
dishes
5.measurer
6.sprayer
7. hair dryer (or oven)
8.layering filter paper (Xinhua One)
9.straight ruler and pencil
(ii) reagents
1. The extender (water-saturated orthotinol and acetic acid mixture)
mixes orthanol and acetic acid in a fractional funnel with a volume ratio of 4:1, and the resulting mixture is mixed with distilled water by volume ratio of 5:3;
2. Amino acid solutions
0.5% lysine, proline, limeine and their mixtures (0.5% for each
). 3. Colorants
0.1% hydrationtridone orthodol solution.
the "Experimental Steps"
. 1. Prepare the filter papertake the lamination filter paper (22cm long, 14cm wide) one sheet, at the edge of one end of the paper 2 to 3cm with a pencil to draw a straight line, in this line each interval of 3cm to mark, as shown in the right image..