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Paper chromatography of amino acids | |||
BAW solvent should be made up fresh on the day.
Using a pencil, lightly draw a line 1.5 – 2cm above the bottom of the chromatography paper. Make small marks at 1.5cm intervals along the line.
Fill a capillary tube or pipette by capillary action with your first sample. Touch the tip of the capillary to the first mark and pull it away. You should leave a small (>5mm) wet spot. Let this dry and apply the sample again. Make a note of which sample is placed on the spot (you can lightly write on the paper beneath the mark, using a pencil). Repeat for each sample or mixture, on a different mark and using a fresh tube or pipette each time. Let the paper dry.
Put 1cm or less of BAW into the chromatography tank. The sample spots should not dip into the solvent. Place the tank in a cool place out of direct sunlight. Stand the chromatography paper in the tank so that the bottom edge is in the solvent but the remaining paper doesn't touch the tank. Place the lid on the tank and leave for about 1 hour.
Wearing gloves, remove the damp paper and mark where the solvent has reached. Dry the paper in the fume cupboard. Still in the fume cupboard, spray the whole of the paper with ninhydrin[?] in butanol. Dry the paper and transfer it to the drying oven for 1-2min.
Mark the position of each spot that develops. Note the colour and Rf (or travel distance) of each standard and hence find the amino acid composition of the unknown samples.
The teeming autumn, big with rich increase,
Like widow'd wombs after their lords' decease:
But hope of orphans, and unfather'd fruit;
And, thou.html">thou.html">thou.html">thou.html">thou away, the very birds.html">birds are mute:
That leaves look pale, dreading the winter.html">winter's near.
XCVIII
From you have I been absent in the spring,
Hath put a spirit of youth in every.html">every thing,
Yet nor.html">nor.html">nor the lays of birds, nor the sweet smell
Could make me any summer's story tell,
Nor did I wonder at the lily.html">lily's white,
They were but sweet, but figures of delight,
Yet seem'd it winter still, and you away,
Sweet thief, whence didst thou steal thy sweet that smells,
Which on thy soft cheek for complexion dwells
The lily I condemned for thy hand,
The roses fearfully on thorns did stand,
A third, nor red nor white, had stol'n of both,
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More flowers I noted, yet I none could see,
To speak of that which gives thee all thy might?
Darkening thy power to lend base subjects light?
In gentle numbers time so idly spent;
And gives thy pen both skill and argument.
If Time have any wrinkle graven there;
And make time's spoils despised every where.
So thou prevent'st his scythe and crooked knife.
CI
O truant Muse what shall be thy amends
Both truth.html">truth and beauty on my love.html">love depends;
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Beauty no pencil, beauty's truth to lay;
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I love not less.html">less, though less the show appear;
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