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# I predict that the juice will have at least 2 or more of the same amino acids as the ones in the amino acid mix. I predict that the amino acid more likely to occur is aspartic acid, as lemon juice is acidic.

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Introduction

CHROMATOGRAPHY Hypothesis I predict that the juice will have at least 2 or more of the same amino acids as the ones in the amino acid mix. I predict that the amino acid more likely to occur is aspartic acid, as lemon juice is acidic. Method * We first extract some lemon juice from a lemon. Cut the lemon around the equator and squeeze it with a juicer. * Pour the juice into six tubes at approximately the same level each. * Place the tubes in a beaker and weigh them (remembering to tare the beaker first). Make pairs of tubes of equal mass by adding or taking out juice with a clean pipette) and place them opposite each other in the centrifuge and set it to run for 5 minutes. * After centrifuging the juice, pour all the supernatant into a beaker and wash out the solid precipitate from the tubes. * Take 2 samples of the raw centrifuged juice in tubes, label and put them away. * Add alcohol to the remaining juice with the ratio of 1:3 (1unit of juice to 3 units of alcohol). ...read more.

Middle

1st aa: x = 9/154 = 0.058 y = 15/154 = 0.097 z = 1/154 = 0.0065 2nd aa: x = 34/154 = 0.22 y = 43.5/154 = 0.28 z = 21/154 = 0.14 Amino Acid Mix 1st aa: x = 33/154 = 0.21 y = 44.5/154 = 0.29 z = 15.5/154 = 0.10 2nd aa: x = 54.5/154 = 0.39 y = 73/154 = 0.47 z = 48.5/154 = 0.31 3rd aa: x = 80/154 = 0.52 y = 85.5/154 = 0.56 z = 74/154 = 0.48 4th aa: x = 104/154 = 0.68 y = 117.5/154 = 0.76 z = 90/154 = 0.58 Asp. x = 35/154 = 0.23 y = 45.5/154 = 0.30 z = 17/154 = 0.11 Leu. x = 77/154 = 0.50 y = 114/154 = 0.74 z = 60.5/154 = 0.39 Lys. x = 52.5/154 = 0.34 y = 72/154 = 0.47 z = 45/154 = 0.29 Pro. x = 75/154 = 0.49 y = 90/154 = 0.58 z = 64.5/154 = 0.42 Percentage Variation (PV) Juice 1st aa: 20-10/10*100 = 100% 0-10/10*100 = -100% PV = 0.065 +- 100% 2nd aa: 46-33.5/33.5*100 = 37.3% 28-33.5/33.5*100 = -16.4% PV = 0.22 + 37.3% or 0.22 - 16.4% Juice + ...read more.

Conclusion

I had marked what I thought was the darkest region of the spot with an 'x.' However that mark was aligned with the mark for Pro and so indicating that they are the same amino acid. This is impossible, but may have been caused by either: Misinterpreting the darkest region of the spot (marking the wrong place) Accidentally using the same micropipette for two substances It would make more sense if the mark on the spot for Leu was closer to the top of the spot, where it would be aligned to one of the spots in the amino acid mix. The 1st amino acid spot for the juice had a smaller spot within it. The smaller spot even showed up as a different color to that of the larger spot, so it must be just another amino acid, not present in the amino acid mix. I could have improved this experiment by making sure that I do not add too much of a substance onto the chromatogram, by getting a second opinion on whether I had marked the darkest region of a spot or not and/or by using a wider variety of amino acids to identify the other amino acids in the juice which do not align with any others. ...read more.

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