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Journal of Drug Delivery and Therapeutics

Open Access to Pharmaceutical and Medical Research

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Open Access Full Text Article  Research Article

Pharmacognostic, Physicochemical, Phytochemical evaluation & Thin layer chromatography of Ocimum basilicum leaves

Brij Raj Singh *

Department of Pharmacognosy, Malti Memorial Trust CSM Group of Institutions, Faculty of B.Pharmacy, 8^th Mile stone Rewa Road, Iradatganj, Prayagraj 212111, U.P., India 

 

 

INTRODUCTION 

Ocimum genus has around 50-150 types of plants¹ and belonging to the Lamiaceae family. Tropical regions are really places to find Ocimum species. These places include Asia, Africa and India.² The Ocimum species are found in these areas. Sweet basil, or Ocimum basilicum, is a plant of the genus Ocimum that is found in South America, Africa, and Asia.³ O. basilicum has large green leaves, glabrous, woody stems, and a broad, elliptical shape that measures 2.5 to 5 cm × 1 to 2.5 cm. It grows to a height of 20 to 80 cm. The 3 mm red, pink, or white flowers are grouped in terminal spikes.⁴ In the past, O. basilicum fruit was used as traditional medicine to treat eye conditions, diarrhea, worm infestation, and inflammation.⁵ In addition to being used as a tonic and vermifuge, leaves and blossoms can be used as a tea to cure diarrhea, gas, and nausea. O. basilicum has essential oils that are frequently used to cure wasp stings, snakebites, colds, and convulsions.⁶ O. basilicum was utilized in Mediterranean cuisine, including Greek and Italian, throughout South Europe.⁷

MATERIALS AND METHODS

Plant collection: Fresh Plant leaves of O. basilicum were collected from local area of Prayagraj, U.P., India. The plant material identified and authenticated by Botanical Survey of India (BSI), Prayagraj, Authentication No.11/81/2025-26/ 54/Tech/912 on dated 30/03/2026.

Macroscopic studies: Organoleptic characteristics, such as color, taste, texture, shape, and size, were used in morphological investigations and assessed botanically.^{8, 9, 10}  

 Microscopic studies: ^{8, 9, 10}

Leaf was cut into tiny transverse slices for microscopic examination. For additional observations and the identification of particular microscopic diagnostic features, the slices were placed in a glycerine water solution. Additionally, the powder properties and fluorescence examination were performed.

 

Physicochemical parameters ^{11, 12}

After powdering the dried plant leaves of O. basilicum, several physicochemical analyses were performed and values were noted, including those for foreign matter, moisture content, extractive value, and ash value. 

Phytochemical screening ^{13, 14, 15}

100g of O. basilicum powder in a Soxhlet extractor was extracted using 95% ethanol. Following the concentration of the liquid extract, a number of qualitative chemical assays were performed and documented.

Total Flavonoids Content¹⁶

First, fill a graduated test tube with 50 ml of the extract, add 1 ml of methanol (Sigma Aldrich, CAS no. 67-56-1), and thoroughly mix. Next, add 4 ml of distilled water and 5% sodium nitrite (CDH, CAS no. 7632-00-0), and incubate for 5 minutes. Let it stand for ten minutes after adding the 10% w/v aluminum chloride solution (CDH, CAS no. 7446-70-0). Next, add 2 ml of 1M sodium hydroxide ( Ranchem, CAS no. 1310-73-2) and use distilled water to get the content to 10 ml. A UV visible spectrophotometer is used to measure absorbance at 765 nm. The standard quercetin (Research Lab fine Chem, CAS no. 849061-97-8) absorbance is determined using the same approach. The calibration curve was used to calculate the total flavonoids content.

Total Phenolic Content¹⁶

Total phenolic content has been determined by FC method. One milligram per milliliter of the extract was put in a test tube; three milliliters of water was put in; then 0.5 milliliters of Folin-Ciocalteu reagent (CDH, batch no. 611221) was mixed. Then, added 20% sodium carbonate (Thomas Baker, CAS no. 497-19-8) and left it in the dark for about 10 minutes. Finally, measured the absorbance at 650 nm with a UV-visible spectrophotometer Likewise, measured the absorbance of gallic acid (CDH, CAS no. 5995-86-8).

Thin layer chromatography ^{17, 18}

Using an activated silica gel G plate as the stationary phase and chloroform: Methanol: Acetic acid (6:3:1) used as mobile phase, and an iodine solution or chamber as the detecting reagent, an alcoholic extract of O. basilicum was assessed for TLC.

RESULTS AND DISCUSSION 

Macroscopic study: The organoleptic & macroscopic characters of the fruits as colour, odour, taste, shape, size, & surface were evaluated botanically.

Figure 1: Ocimum basilicum plant

 

 

Table 1: Macroscopic study of Ocimum basilicum leaf

Characters	Leaf
Colour	Upper surface darkish green upper surface and light green lower surface
Odour	Pleasant, aromatic, strong
Taste	Sweet
Size	2.5-5.3 cm long, 1.5- 2 cm width
Shape	Simple, ovate, oval
Margin	Entire, smooth
Apex	Acute
Venation	Pinnate, Reticulate
Surface	Pubescent
Base	Attenuate

 

 

 

 

Microscopic study:

Transverse section of O. basilicum leaf shows single layered cylindrical palisade cells; 4-6 layers of parenchymal cells, vascular bundles were ringed by parenchymatous cells. Glandular trichomes and diacytic stomata were also present

 

 

 

.                                            

Figure 2: Transverse Section of Ocimum basilicum

 

Table 2: Powder characteristics of Ocimum basilicum leaf

S. no.	Features	Observation
		Leaf
1	Nature	Coarse powder
2	Colour	Greenish
3	Odour	Noxious
4	Taste	Slightly bitter

 

 

Powder microscopy: It is greenish coarse powder with bitter taste. The powder microscopic study reveals the presence of trichomes, parenchymal cells, lignin, calcium oxalate crystal & volatile oil sac.

 

 

Table 3: Fluorescence analysis of Ocimum basilicum leaf powder

Treatment	Visible	Long U.V. 254 nm	Short U.V. 365 nm
Powder	Brown	Black	Green
Powder + % KOH	Dark brown	Dark	Light green
Powder + 5%NaOH	Blackish green	Blackish	Green
Powder + FeCl3	Black	dark	Blackish green
Powder + con. H2SO4	blackish	Dark	Blackish green
Powder + dil. H2SO4	Brown	Dark	Green
Powder + con. HCl	Brown	Black	Dark Green
Powder + dil. HCl	Brown	Dark	Greenish
Powder + con. HNO3	Black	Dark	Light Green
Powder + dil. HNO3	Brown	black	Light green
Powder + dil. NH3	Black brown	Dark	Green
Powder + Iodine soln	Black -brown	Black	Blackish green

 

 

Table 4: Physicochemical data of Ocimum basilicum leaf

S.N.	Physicochemical Parameter	Values (% w/w)
1.	Foreign matter	Nil
2.	Moisture Content	13%
3.	Total Ash	14.60%
4.	Acid- Insoluble ash	4.15%
5.	Water soluble ash	3.27%
6.	Alcohol soluble extractive	2.4%
7.	Water soluble extractive	1.6%

 

Percentage yield and physical characteristics of various extract of O. basilicum leaf 

Solvent extract	%w/w	Consistency	Fluorescence analysis
			Visible	Long U.V.	Short U.V
Ethanol (95%)	28.38	Sticky	Blackish brown	Dark black	Greenish

 

 

Qualitative phytochemicals screening

Phytochemical screening of ethanolic extract of O. basilicum shows the presence of several secondary metabolites. Shinoda and zinc-hydrochloric acid tests shows the presence for flavonoids while triterpenoids & alkaloids were absent. Saponins, tannins, and steroids were also present.

 

Table 5: Qualitative Phytochemical screening of Ethanolic extract of O. basilicum

	Phytochemical test	Ethanolic extract of O. basilicum
1.	Carbohydrates i) Molisch ii) Fehling Reagent	-ve -ve
2.	Alkaloids i) Dragondroff’s reagent ii) Mayer’s reagent iii) Wagner reagent iv) Hager reagent	- - - -
3.	Tannins i) Lead acetate ii) FeCl3	+ +
4.	Flavonoids i) Shinoda test ii) Zinc- HCl reduction test iii) Alkaline reagent test	++ ++ ++
5.	Saponins i) Foam test	+
6.	Steroids i) Libermann – Burchard test	++
7.	Cardiac glycosides Keller-Kiliani	+
8.	Triterpens Salkowaski’s test	-

     +++ More amount, ++ moderate amount, + less amount, - absent   

                                                                                             

 Table 6: Total Phenolic Content and Total Flavonoid content

Extract	Total Phenolic content mg GAE/g	Total Flavonoid content mg Quercetin equivalent/g
Ethanolic extract O. basilicum leaves	25.40	18.30

 

Table 7: TLC Profile: Thin layer chromatography of alcoholic extract of O. basilicum

Solvent system	No. of spots	Rf value
Chloroform: Methanol: Acetic acid (6:3:1)   Spraying agent/Detection : Iodine chamber	3	0.60, 0.68, 0.72

 

 

DISCUSSION:

T.S. of O. basilicum shows isodiametric epicarp with thin cuticle, mesocarp have 5-7 layers of large parencymatous elongated cells with starch grains, vascular bundles are present in the mesocarp. Powder microscopy reveals the presence of parenchymal cells, trichomes, & calcium oxalate crystals. Qualitative phytochemical screening indicates presence of alkaloids, tannins, tritepenes, steroids and flavonoids, carbohydrates; total phenolic content and total flavonoids contents were found 25.40 mg GAE/g and 18.30mg Quercetin eq/g respectively; thin layer chromatography of ethanolic extracts indicates presence of many compounds. 

CONCLUSION:

The plant O. basilicum is a common species that has been used traditionally. The above pharmacognostic, physicochemical, phytochemical & chromatographic studies will give approaches for identification, safety & quality parameters as well as new incentive to natural system of medicine in the research & in the treatment of other diseases.

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