Procedure I - Thin-Layer Chromatography

Using the lipids key for lanes 1-3 found under the Activity Form tab, list the lipids present in lanes 4 and 5 of the chromatography plate in the table below. List the lipids using the abbreviations found under the Activity Form tab (TG for triglycerides, etc.).

Tip: It is helpful to use a ruler when attempting to match up lipids on the chromatography plate.

Table I-1. TLC Plate Bands for Fish and Nut Samples
Lane 1 Standards	Lane 2 Standards	Lane 3 Standards	Lane 4 Fish	Lane 5 Nut
CE
	TG
	DHA
	EPA
	ALA
	LA
	OA
	PA
FC
		PS

Observations and Questions

[1] Closely examine the bands for the fish and nut samples. Describe the overall appearance (number, intensity, and distribution) of the bands for each test sample, the fish and nut samples.

[2] How many bands do the fish and nut samples have in common?

[3] Compare the standard bands with the bands on the TLC plate that the two test samples have in common. Identify (name) the lipids that the fish and nut samples have in common. Be specific and refer to your results in Table I-1 above.

[4] For each of the fatty acids that the fish and nut samples have in common, discuss which sample (fish or nut) contains the greater amount of those fats. Hint: Refer back to your Procedure I screenshot and note that the darkness of a band is an indication of the amount.

[5] Which lipids are distinct, i.e., not in common, for the fish and nut samples? In other words, how is the lipid composition different between the fish and nut samples? Be specific and refer to your results in Table I-1 above.

[6] Omega-3 fatty acids are associated with reductions in the risks of cardiovascular disease, reduced inflammation, and a healthy balance of LDL and HDL cholesterol. Based on your results, can all three omega-3 fatty acids be obtained from fish? Or from nuts? Discuss information from your Table I-1 results and your TLC plate to explain how you arrived at your answer.

Procedure II - Unsaturation Testing

In the table below indicate the number of drops needed for each sample to change color.

Table II-1. Unsaturation Test for Presence of Double Bonds
Drops Needed for Sample A	Drops Needed for Sample B	Drops Needed for Sample C	Drops Needed for Sample D	Drops Needed for Sample E

Observations and Questions

[7] Based on information you learned from the Background material, what does the test of unsaturation reveal about the structural biology of lipids?

[8] Think about your experience dispensing the drops in this experiment. What is the nutritional significance of the ability of the lipid samples to retain the color of the solution in the dropper? For example, how does the colorization process observed here help you to understand which lipids should be more abundant in your diet and which should be less abundant?

[9] List the fatty acid samples (A-E) from lowest to highest saturation. Label the lowest and highest. Discuss how knowledge of, and experience with, the lipid saturation test can be used to make decisions about the saturation ranks. BE CAREFUL: This question is asking about 'saturation.'

LOWEST SATURATION

:

:

:

HIGHEST SATURATION

[10] Based on your Procedure II results (Table II-1), identify which fatty acid sample (A-E) corresponds to each fatty acid (and its corresponding number of double bonds) given in Table II-2 below.

Table II-2. Test Sample-to-Fatty Acid Family Member Correspondence
Fatty Acid	Number of Double Bonds	Fatty Acid Sample	Omega-# Fatty Acid Identification
DHA	6
EPA	5
ALA	3
LA	2
OA	1

[11] The fatty acid acronyms listed in Table II-2 all belong to the omega fatty acid family. Conduct research or an internet search to determine the corresponding omega-# identification for each of the five fatty acids in the table. Add the corresponding names to the rightmost column of Table II-2.

[12] Choose one of the omega fatty acid family members you listed in Table II-2 to address questions (a) and (b) below. Cite all sources used to answer the question.

(a) What are the known health benefits associated with including this member of the omega fatty acid family as a regular part of your dietary intake?

(b) Is the omega fatty acid member that you selected regarded as an essential fatty acid? Explain your reasoning.

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