**3.3. Qualitative screening of saponins**

The general method involved in the qualitative analyze of saponins is: 2 ml of aqueous extract and 2 ml of distilled water are shaken for 15 min in a graduated cylinder. A 1 cm foam layer is a positive response to the presence of saponins (see **Table 6**).

Qualitative screening of saponins in aqueous extracts from sea buckthorn and gooseberry revealed that only the second one contains saponins.

The test for phenolic flavonoids (**Figure 7**): 1 ml aqueous extract is mixed with 2 ml of 10% lead acetate solution and a brown precipitate indicates a positive response (see **Table 7**).

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Flavonoids are present in both aqueous extracts (sea buckthorn and gooseberry), while phe-

Alkaloids are a group of basic plant bioactive compounds that possess an N-containing heterocycle, are generally colorless, crystalline, insoluble in water but soluble in many organic solvents [29].

nolic flavonoids are present as small traces in gooseberry aqueous extract.

**Phytochemical test Sea buckthorn Gooseberry**

**Phytochemical test Sea buckthorn Gooseberry** Saponins 0.2 cm foam layer 1.5 cm foam layer

Tannins Green-black solution Green-black solution Phlobatannins Pale pink solution Red-orange solution

**3.5. Qualitative screening of alkaloids**

**Figure 7.** Phenolic flavonoids in sea buckthorn and gooseberry.

**Table 6.** Qualitative screening of saponins.

**Table 5.** Qualitative screening of tannins and phlobatannins.

**Figure 6.** Hexose sugars in sea buckthorn aqueous extract.

### **3.4. Qualitative screening of flavonoids and phenolic flavonoids**

Flavonoids have important functions in plants: attract pollinating insects, fight against different microbial infections, and control cell growth [28].

Flavonoids are tested according to the following method: 2 ml aqueous extract and 1 ml of 2N sodium hydroxide are mixed. A yellow color indicates the presence of flavonoids.


**Table 4.** Qualitative screening of hexose sugars.

Qualitative Analysis of Phytochemicals from Sea Buckthorn and Gooseberry http://dx.doi.org/10.5772/intechopen.77365 169

**Figure 6.** Hexose sugars in sea buckthorn aqueous extract.

**c.** *Ammonium molybdate test*: this test reveals the presence of ketohexozes as follows: to 2 ml aqueous extract, 2 ml ammonium molybdate solution are added, the solution is then heat-

As it is clear from the **Table 4**, hexose sugars are present in both sea buckthorn aqueous

Most of the tannins, a group of phenol compounds usually found in plants, are soluble in water. Phlobatannins are considered a novel class of ring-isomerized condensed tannins [17]. The test for tannins is generally described as [27]: to 1 ml aqueous extract 2 ml of 5% ferric

Phlobatannins are tested following a standardized method: to 1 ml aqueous extract of sea buckthorn and gooseberry few drops of diluted HCl (1%) is added and a red precipitate

Tannins are present in both aqueous extracts, while small traces of phlobatannins can be

The general method involved in the qualitative analyze of saponins is: 2 ml of aqueous extract and 2 ml of distilled water are shaken for 15 min in a graduated cylinder. A 1 cm foam layer

Qualitative screening of saponins in aqueous extracts from sea buckthorn and gooseberry

Flavonoids have important functions in plants: attract pollinating insects, fight against differ-

Flavonoids are tested according to the following method: 2 ml aqueous extract and 1 ml of 2N

Lower layer-blue precipitate, upper

Cognac-red solution Red solution

Blue-green solution Blue-green solution

Reddish solution, yellow-white

precipitate

sodium hydroxide are mixed. A yellow color indicates the presence of flavonoids.

layer-pink solution

**Phytochemical test Sea buckthorn Gooseberry**

ed to form a bluish-green solution.

should appear (**Table 5**).

Carbohydrates (hexose sugars)—Seliwanoff

chloride

Carbohydrates (hexose sugars)—cobalt

**Table 4.** Qualitative screening of hexose sugars.

Carbohydrates (hexose sugars) ammonium molybdate

found in gooseberry aqueous extract.

**3.3. Qualitative screening of saponins**

extract as well as in gooseberry aqueous extract.

168 Phytochemicals - Source of Antioxidants and Role in Disease Prevention

**3.2. Qualitative screening of tannins and phlobatannins**

chloride are added and a dark-blue or greenish-black color appears.

is a positive response to the presence of saponins (see **Table 6**).

**3.4. Qualitative screening of flavonoids and phenolic flavonoids**

revealed that only the second one contains saponins.

ent microbial infections, and control cell growth [28].


**Table 5.** Qualitative screening of tannins and phlobatannins.


**Table 6.** Qualitative screening of saponins.

The test for phenolic flavonoids (**Figure 7**): 1 ml aqueous extract is mixed with 2 ml of 10% lead acetate solution and a brown precipitate indicates a positive response (see **Table 7**).

Flavonoids are present in both aqueous extracts (sea buckthorn and gooseberry), while phenolic flavonoids are present as small traces in gooseberry aqueous extract.

#### **3.5. Qualitative screening of alkaloids**

Alkaloids are a group of basic plant bioactive compounds that possess an N-containing heterocycle, are generally colorless, crystalline, insoluble in water but soluble in many organic solvents [29].

**Figure 7.** Phenolic flavonoids in sea buckthorn and gooseberry.


According to the results presented in **Table 9**, anthraquinones and anthocyanosides are absent

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Proteins are involved in all physiological processes that take place in all living cells. Proteins are colloidal, do not diffuse through the plasma membrane, are irreversible coagulated upon

Amino acids are amphoteric phytocompounds, highly reactive, with an amino and carboxylic

Experimental: 1 ml aqueous extract reacts with 5–6 drops of Millon's reagent (mixture of mercuric nitrate, mercurous nitrate, concentrated nitric acid, and distilled water) and a white precipitate is formed that changes its color to red upon heating. Millon's test is a non-specific test for detecting proteins and amino acids (tyrosine) and, therefore, it must be confirmed by other qualitative tests. The results obtained after the two aqueous extracts react with Millon reagent are as follows: an opalescent orange solution in the case of Sea buckthorn and a red-brownish precipitate in the case of Gooseberry, therefore confirming the presence of small amounts of proteins and/

**a.** *Ninhydrin test*: take 3 ml aqueous extract and mix it with three drops of 5% lead acetate solution then heat the resulted solution. A purple or blue coloration indicates a positive

**b.** *Test for cysteine*: 5 ml aqueous extract is boiled with a small amount of 40% NaOH and few

The test for cysteine gives a positive reaction in the case of sea buckthorn, while ninhydrin test

precipitate

Opalescent dark-brown solution

drops of 5% lead acetate solution are added. A black precipitate is formed.

Proteins and amino acids—Millon Opalescent orange solution Red-brownish precipitate

Amino acids—ninhydrin test Opalescent white-yellow solution Opalescent orange solution, gray

from both aqueous extracts.

**3.7. Qualitative screening of proteins and amino acids**

heating and are insoluble in neutral salts [30].

acid moiety, therefore, being mostly water soluble.

*3.7.1. General screening of proteins and amino acids*

or aminoacids in Gooseberry aqueous extract.

There are two different standard methods used (see results in **Table 10**):

**Phytochemical test Sea buckthorn Gooseberry**

precipitate

*3.7.2. Detection of amino acids*

reaction (**Figure 8**).

is negative for both aqueous extracts.

**Table 10.** Qualitative screening of amino acids.

Amino acids—test for cysteine Red-brown solution, black

**Table 7.** Qualitative screening of flavonoids and phenolic flavonoids.

There are three different standard phytochemical methods used to determine the presence of tannins in aqueous extracts from sea buckthorn and gooseberry:


According to the results presented in **Table 8**, alkaloids are absent from all the aqueous extracts.
