Feb 27, 2023
Preliminary qualitative phytochemical analysis
- Matin Mahmood1,
- Abdulkareem Hameed Abd2,
- Enas Jawad Kadhim3
- 1Department of Pharmacology, College of Pharmacy, Alkitab University, Altun Kopre, Kirkuk, Iraq;
- 2Department of Pharmacology, College of Medicine, Al-Nahrain University, Kadhimiya, Baghdad, Iraq;
- 3Department of Pharmacognosy and Medicinal Plants, College of Pharmacy, University of Baghdad, Baghdad, Iraq
Protocol Citation: Matin Mahmood, Abdulkareem Hameed Abd, Enas Jawad Kadhim 2023. Preliminary qualitative phytochemical analysis. protocols.io https://dx.doi.org/10.17504/protocols.io.e6nvwj27wlmk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: February 26, 2023
Last Modified: February 27, 2023
Protocol Integer ID: 77638
Keywords: Detection of fractions, Phenolics, Terpene
Abstract
Plant extraction is a process that aims to extract certain components present in plants. It is a solid/liquid separation operation: a solid object (the plant) is placed in contact with a fluid (the solvent). The plant components of interest are then solubilized and contained within the solventChemical tests were carried out using the ethanolic extracts from plants and different fractions, using standard procedures to identify the active constituents
Materials
I.Alkaloids test:
HCL (Sigma-Aldrich cat-no. 1009861000)
Mayer’s (1.35gm mercuric chloride (Sigma-Aldrich, Germany ca-no. 215465) in 60ml water + 5gm potassium iodide (Sigma-Aldrich, Germany cat-no. 221945) in 10ml water)
Wagner’s reagents (1.27g of iodine (Sigma-Aldrich, Germany cat-no. 1047630050)
II.Flavonoids tests
a.Lead acetate test: lead acetate solution (BDH limited, England cat-no. LL0093)
b.NaOH test: NaOH and HCl
III.Steroids tests
sulfuric acid (BDH limited, England cat-no. BDH3068-500MLP)
IV. Terpenoids test (Salkowski test):
chloroform (Honeywell, USA cat-no. C2432)
sulphuric acid (BDH limited, England cat-no. BDH3068-500MLP) w
Test for alkaloids
Test for alkaloids
Ten (2ml) of alcoholic extract and fractions were stirred with 5 ml of 1% HCL on a steam bath.
Mayer’s (1.35gm mercuric chloride in 60ml water + 5gm potassium iodide in 10ml water )and Wagner’s reagents (1.27g of iodine and 2g of potassium iodide in 100ml of water) were added, white and reddish brown color precipitate respectively, were taken as evidence for the presence of alkaloids
Test for flavonoids
Test for flavonoids
Two tests used for detection of flavonoids
Lead acetate test: to 2ml of alcoholic extract and fractions, 1ml of 10% lead acetate solution was added. The formation of a yellowish- white precipitate was taken as a positive test for flavonoids
NaOH test: 2ml of the extract and fractions were treated with aqueous NaOH and HCl, and looking for the formation of a yellow orange color
Tests for steroids
Tests for steroids
Two tests used for identification of steroids
Liebermann-Burchard test: 2ml of the extract and fractions were treated with chloroform, acetic anhydride and drops of sulphuric acid was added. The formation of dark pink or red color indicates the presence of steroids
H2SO4 test: The development of a greenish color was considered as indication for the presence of steroids, when 2 ml of the extract and fractions were treated with sulphuric and acetic acids.
Test for terpenoids
Test for terpenoids
Two ml of the organic extract and fractions were dissolved in 2 ml of chloroform
Evaporated to dryness
2 ml of concentrated sulphuric acid was then added and heated for about 2 min. A grayish color was considered an indication for the presence of terpenoids.