FEHLING’S TEST

Fehling’s test is used in detecting reducing sugars in a sample. The Fehling’s reagent is composed of two
solutions: Fehling’s A and Fehling’s B. In this test, to confirm that the sample is positive the substance
should show reddish brown precipitate such as glucose, fructose, and lactose (Caton, 2011). Based from
the table above, all of the samples turned into brick red colored substance except for starch and
glycogen. Samples like glucose, xylose, fructose, maltose, and unknown sugar resulted to a positive
substance (brick red color), thus they are reducing sugars. The presence of brick red precipitate was due
to the Fehling’s Reagent containing Cupric ion (Cu2+). Cupric ion then is reduced to Cuprous ion (Cu+)
therefore, forming a red precipitate of Cu2O. On the other hand, starch and glycogen are non-reducing
sugars. Non-reducing sugars do not have an OH group attached to the anomeric carbon so they cannot
reduce other compounds (Gunawardena, 2011). As for starch, the first ring cannot open up because
there is no hydrogen to allow for ring opening. Glycogen has a lot of branches thus it gives a lot more
ends for enzymes to work on to hydrolyze the glycogen to form glucose. Starch and glucose have to be
hydrolysed and broken down into smaller units before the rings can open up.

CHEM EQ:

(n.d.). Retrieved from http://dl.clackamas.edu/ch106-07/disaccha.htm

Caton, K. A. (2011, July 15). Carbohydrates. Retrieved from

https://www.slideshare.net/katealyssacaton/carbohydrates-8609628

Gunawardena, G. (2019, June 6). Nonreducing sugars. Retrieved from

https://chem.libretexts.org/Ancillary_Materials/Reference/Organic_Chemistry_Glossary/Nonreducing_S
ugar
BARFOED’S TEST

Aldoses and ketoses can reduce cupric ions even in acidic situations. The Barfoed’s Test is used to detect
the presence of monosaccharides (reducing sugars) in a solution (“Objective,” n.d). Based on the table
above, only xylose and lactose had a change in color surface. The rest of the samples did not show any
presence of reducing sugars. However, the above result is faulty because of human error – the time for
observing the possible changes was not followed. It was instructed that after heating the substance for
30 seconds in a boiling water bath, it should be allowed to stand for 15 minutes before noting the
results. Due to time scarcity, it was only allowed to stand for approximately 8 minutes. Nevertheless,
results should be positive in glucose, fructose, and xylose. In a positive result a scanty brick red
precipitate should be observed. In monosaccharides, the changes were noticed in a shorter amount of
time because monosaccharides are stronger reducing agents compared to disaccharides. Reducing
monosaccharides are oxidized by the copper ion in solution to form a carboxylic acid and reddish
precipitate of copper (I) oxide within three minutes. Due to the weakly acidic nature of Barfoed's
reagent, it is reduced only by monosaccharides.

Objective. (n.d.). Retrieved from https://vlab.amrita.edu/?sub=3&brch=63&sim=631&cnt=1

TOLLEN’S TEST

Tollen’s test is used to distinguish between an aldehyde and a ketone (Libretexts, 2019). The tollen’s
reagent is colourless, basic, and aqueous solution containing silver ions. The expected result for this test
is that there should be a silver mirror on the innerside of the test tube. As for the table above, the actual
result is totally different from the expected result. According to the table above, the result gave a
brownish precipitate. However, there came a problem. The H2O that was used in immersing the tollen’s
reagent was not warm enough to have the substance show the expected result. The substance was
immersed in warm H2O for approximately 3 minutes only because of time scarcity. Nonetheless, the
silver mirror that is expected to appear in the substance is due to the oxidation of aldehyde into the
corresponding carboxylic acid. The reaction is accompanied by the reduction of silver ions in Tollen’s
reagent into metallic silver, thus, forms a mirror on the test tube.

Libretexts. (2019, June 05). Tollens' Test. Retrieved from

https://chem.libretexts.org/Bookshelves/Organic_Chemistry/Supplemental_Modules_(Organic_Chemist
ry)/Aldehydes_and_Ketones/Reactivity_of_Aldehydes_and_Ketones/Tollens’_Test

What happens when glucose reacts with Tollen's reagent? (n.d.). Retrieved from
https://www.quora.com/What-happens-when-glucose-reacts-with-Tollens-reagent

ANSWER TO QUESTION 1:

Molisch’s Test is a general test for all carbohydrates – monosaccharide, disaccharides, and
polysaccharides (Karki, 2018). In this test, monosaccharides are dehydrated in the presence of
concentrated sulfuric acid to form an aldehyde as furfural (pentoses) or hydroxymethyl
(hexoses) derivatives. Disaccharides and polysaccharides are converted to monosaccharide
through hydrolysis of the glycosidic bonds. It is the responsibility of the acid to hydrolyse the
carbohydrates into monosaccharides especially if it is either disaccharide or polysaccharide.
The derivatives then react with 1-naphthol/alpha-naphthol in Molisch reagent through a
condensation reaction to form purple colored compound on the junction of the sample.
ANSWER TO QUESTION 4:

Strong alkalies decompose the sugars. Weak alkalies, like sodium bicarbonate, act upon the
sugar. The monosaccharides are easily decomposed by alkalies, even very weak ones.