Thin Layer Chromatography Procedure

1) Apparatus:-
Ultraviolet light (UV), glass plates, 4×8, spreader , oven ,micro pipette, beaker ,shaker ,electric balance, water bath, china dish
2) Reagents:-
Silica gel (powder form) 254 Gr 60 number, double dish H2O, chloroform, standard solution,
3) Procedure:-
Sample preparation:-
a) Take 25 gm sample add 20 ml distilled water.
b) Mix it and add 100ml chloroform in it.
c) Shake for 15 minutes.
d) And filter it in china dish put china dish in water bath and dried the solution and then cool it.
e) Add 1ml chloroform and make spots of 2, 5, 7 and 10 micro liter.
f) Make spots of standard solution with micro pipette.
Preparation of silica gel plates:-
a) Take silica gel powder form 254 Gr 60 number and add double distilled water in it put 2.5 to 3 gm silica gel on spreader and spread it on plate.
b) Stay it as such for some time and then transfer it in oven at 100C for 2 hours. OR 60 to 70C for one night.
c) Next day make spotting on it. And then calculate the toxin with the help of standards.
Solution Preparation:-
1) Standards of sodium:-
23+35.5=58.5/23=2.543+distilled water = 1000ml 1gm/1000ml or 1gm/1liters or 1mg/ml
Take 10ml from this + distilled water = 100ml
Now take 20ml from it (100ml) and + add distilled water= 100ml it is 20ppm
Now take 10ml from 100ml and + add water= 100ml it is 10ppm
Now take 60ml from 100ml and + add water= 100ml it is 60ppm
Now take 80ml from 100ml and + add water= 100ml it is 80ppm
2) For Potassium:-
K2SO4 = (039 x2) +32+64=174/78=2.231gm + Distilled water 1000ml other procedure is same as above.