Journal of Biotechnology and Phytochemistry

All submissions of the EM system will be redirected to Online Manuscript Submission System. Authors are requested to submit articles directly to Online Manuscript Submission System of respective journal.
Reach Us +44 7460731551

Research Article - Journal of Biotechnology and Phytochemistry (2022) Volume 6, Issue 4

Phytochemical analysis and anti inflammatory activity in aqueous and hydro ethanolic extract of Morinda citrifolia leaves.

D Chandra Prabha*, Nivetha V

Department of Biochemistry, Sri Ramakrishna College of Arts and Science for Women, Tamil Nadu, India

Corresponding Author:
Dr. D Chandra Prabha
Department of Biochemistry
Sri Ramakrishna College of Arts and Science for Women
Tamil Nadu
E-mail: [email protected]

Received: 24-February-2021, Manuscript No. M-26761; Editor assigned: 01-March-2021, PreQC No. P-26761; Reviewed: 15-March-2021, QC No. Q-26761; Revised: 31-May-2022, QI No. Q-26761 Manuscript No. R-26761; Published: 28-June-2022, DOI: 10.35841/aajbp-6.4.116.

Citation: Prabha DC, Nivetha V. Phytochemical analysis and anti inflammatory activity in aqueous and hydro ethanolic extract of morinda citrifolia leaves. J Biotech Phytochem. 2022;6(4) :116.

Visit for more related articles at Journal of Biotechnology and Phytochemistry


The Morinda citrifolia commonly called as Indian noni or Indian mulberry or Manjanathi is widely known for its therapeutic effect. Manjanathi belongs to the family Rubiaceae. It contains minerals such as selenium and zinc. In this study, we made an attempt to investigate phyto compounds and anti-inflammatory activity in aqueous and hydroethanolic extract of fresh leaves of the manjanathi (Morinda citrifolia). Leaves were extracts by the standard  protocol. The fresh leaves were shade dried and extracted with water and hydroethanol. The aqueous extract showed positive results for alkaloids, cardiac glycosides, terpenoids, saponin, carbohydrates and reducing sugar, phlobatanins and quinone. Whereas the hydroethanol extract showed the presence of alkaloids, tannin, terpenoids, carbohydrates, cardiac glycosides and reducing sugar. The M. citrifolia plant extract contains the increased anti inflammatory activity in the concentration of 50 µl the aqueous and hydroethanolic extract shows the maximum range is 87.26 and 97.36. It can be concluded that although the bioactive components responsible for the therapeutic effect are extracted in the polar and organic extracts. However further characterization is required to elucidate the mechanism of therapeutic action.


Morinda citrifolia, Aqueous, Hydro ethanol, Phytochemical, Anti-inflammatory.


Medicinal plants play a significant role in providing primary health care services to people and serve as therapeutic agents as well as important raw materials for the manufacture of traditional and modern medicine [1]. The World Health Organization (WHO) reported that 80% of people in the developing world use medicinal plants for their primary health careduring the last 10 years an intense interest has emerged in “nutraceuticals” (or functional foods) in which phytochemical constituents can have long term health promoting or medicinal qualities [2-5].

Morinda citrifolia L. commonly known as Indian Noni, is one of the most significant sources of traditional medicine, belongs to the family Rubiaceae and its fruit has long history of use as a food in tropical regions throughout the world. It is also known as Indian mulberry. The species of Morinda especially M. citrifolia has been reported to have a broad range of health benefits for cancer, infection, arthritis, asthma, hypertension, and pain [6]. Noni has a broad range of therapeutic effects such as analgesic, anti-inflammatory, antihypertensive, immune enhancing, anticancer, antibacterial, antiviral, antifungal, antituberculous, anti-protozoal, antioxidant, anti-stress and also sedative properties. The high therapeutic profile and safety potential of Noni has made it a popular health enhancer and food supplement worldwide [7-9]. Antioxidant and free radical scavenging effect of M. citrifolia fruit extract. Anti diabetic potential of Morinda citrifolia products. Morinda citrifolia L., has been applied to prevention and treatment of various diseases relating to skin, brain, gastrointestinal tract, heart, liver and cancer.

The present study was designed to study the presence of various phytochemical constituents present in the aqueous and hydro ethanolic extract of Morinda citrifolia L by the standard phytochemical tests.

Materials and Methods

Collection of plant materials

The fresh leaves of  Morinda citrifolia were collected, cleaned and shade   dried for two weeks. Then leaves were powdered  by mortor and pestle until to get coarse powder and are stored at refrigerator.

Extraction preparation

12.5 gm of shade dried powdered sample was extracted with 50 ml of aqueous and hydro ethanol and kept for extraction for two days by covering with an absorbent cotton to avoid contamination, then thoroughly shaken intermittently. At the end of the second day the extract was collected, filtered by using muslin cloth and filtrate was taken out  and it maintained at room temperature.

Phytochemical constituents

Phytochemicals screening of the prepared extracts was conducted with various qualitative test to identify the presence of chemical constituents.

Test for alkaloids (Wagner’s test)

Add few drops of diluted Hydrochloric acid and filter the extract and then filtrate the tested for wagner’s reagent (red brown precipitate).

Test for flavonoids (Lead acetate test)

1 ml of test solution was treated with few drops of lead acetate (10%) solution, yellow precipitate is formed presence of flavonoid.

Test for cardiac glycosides (Keller-Killani test)

The 2 ml of extract is added with the 2 ml of chloroform then add the equal volume of  concentratedsulphuric acid to  form a reddish brown colour steroid ring indicates the presence of cardiac glycosides.

Test for anthraquinone

1 ml of extract is boiled with 10% HCl for few minutes in water bath. Its filtered and allowed to 100 ml. Equal volume of  CHCl3 is added to filtrate, few drops of 10% ammonia is added to the mixture and heat. Formation of rose pink color indicates the presence of anthraquinone.

Test for saponin (Foam test)

1 ml of extract was shaken with 2 ml of water. If form produced persist for 10 minutes it confirms the presence of saponin.

Test for tannin (Ferric chloride test)

To 2-3 ml of extract was taken and added 5% of feric chloride, the appearance of brownish green colour indicated the presence of tannins and phenolic compounds.

Test for terpenoids

3 ml of extract were added with 3 ml of chloroform and sulphuric acid  to form a reddish brown colour layer indicates the presence of terpenoids.

Test for proteins and aminoacids

Ninhydrin test: To 1 ml of extract added few drops of  freshly prepared 0.2% Ninhydrin reagent and heated. A blue colour develops indicating the presence of protein.

Xantho protein test: To 1 ml of extract added few drops of  sodium hydroxide. Orange colour indicates presence of amino acids.

Test for carbohydrates (Fehling’s test)

Equal volume of Fehling’s solution A and B was added to 1ml of extract and heat gently. Formation of brick red precipitate  indicates the  presence of carbohydrates.

Test for reducing sugar (Fehling’s test)

1 ml of the extract was added with few drops of Fehling’s reagent. Then the mixture was boiled in a boiling water bath for about 10 minutes to observe the formation of blue colour.

Test for phenols (Ferric chloride test)

1 ml of extract was added with the 1 ml of ferric chloride. Then it form deep blue colour presence of phenols.

Test for phlobatanins

The sample is dissolved with distilled water and filtered. Then the filtrate is boiled with 2% HCl solution to form the red color precipitate indicates the presence of phlobatanins.

Test for quinones

1 ml of the peel extract was added to 1 ml conc. sulphuric acid. Formation of red color indicates the presence of quinones.

In vitro anti-inflammatory activity

Anti-inflammatory activity by inhibition of protein denaturation method was evaluated. The aqueous and hydroethanolic extracts of the plant Morinda citrifolia were added in different concentrations such as 10 μl, 20 μl, 30 μl, 40 μl and 50 μl. To this added 0.45 ml of 1% aqueous solution of bovine serum albumin in all the test tubes. The pH is adjusted by adding a small amount of 1N HCl, this sample were kept in incubation for 20 minutes at 37°C and then heated at 57°C for 20 minutes. Then the samples are allowed to cool and 2.5 ml of phosphate buffer is added in all the tubes. In control tube 0.45 ml of bovine serum albumin, 0.05 ml of distilled water and 2.5 ml of phosphate buffer were added. The turbidity was measured at 660 nm under UV-Visible spectrophotometer. The experiments were carried out in triplets. Aspirin is used as standard. The percentage inhibition of protein denaturation was calculated.

%inhibition=[Abs control-Abs sample]/Abs control × 100]

Results and Discussion

The results of the phyto constituents residing in the aqueous and hydro ethanolic extract of Morinda citrifolia L are tabulated in Table 1.

Phytoconstituents Aqueous extract Hydro ethanolic extract
Alkaloids + +
Flavonoids - -
Cardiac glycosides + +
Anthraquinone - -
Saponin + -
Tannin - +
Terpenoids + +
Protein - -
Amino acids - -
Phlobatanins + -
Carbohydrates + +
Reducing sugar + +
Phenol - -
Quinone + -

Table 1. Phytoconstituents in Morinda citrifolia L.

Different phytochemicals have been found to possess a wide range of activities, which may help in protection against chronic diseases [10]. The aqueous extract showed positive results for alkaloids,cardiac glycosides, terpenoids, saponin, phlobatannins, carbohydrates and reducing sugars and quinone negative results were observed for flavonoids, tannin, anthroquinone, protein, amino acids and phenol [11-15]. Phyto components such as alkaloids, tannin, terpenoids, carbohydrates, reducing sugar and cardiac glycosides were present in the hydro ethanolic extract of M. citrifolia L and Flavonoids, Anthraquinone, Saponin, protein, amino acids, Phlobatanins, Phenol and quinone were absent. The alkaloids protect against chronic diseases. Phyto constituents protect against hypercholesterolemia and possess antibiotic properties. They exhibit the analgesicproperties. Alkaloids especially are capable of reducing headaches associated with hypertension.

From the results of the present study, it is obvious that the hydro ethanolic extract also has several phyto components which indicates that the bioactive principles of M.citrifolia are separated in polar and nonpolar.

Positive tests of alkaloids, tannins and coumarins in the ethanolic extract of leaves of Morinda citrifolia L. The phenols and saponins in ethanolic extract of Morinda citrifolia L. leaves have also been recorded [16]. These are in agreement with our results displaying the availability of alkaloids, tannins, coumarins, phenols and saponins in leaves.

The positive analysis of coumarin glycosides in the ethanolic extract of Morinda citrifolia L. fruits [17]. The positive analysis of phenols, cardiac glycosides and steroids in the ethanolic extract of roots of Morinda citrifolia L. Besides, coumarin tested positive in the ethanolic extract of Morindacitrifolia L. roots, which is similar to the records.

Inhibition of protein denaturation

Protein denaturation is a procedure by which the utilization of outer pressure or intensify the organic proteins lose their optional and tertiary structure. At the point when denaturation happens the majority of the organic proteins lose their natural capacity.  It is critical to demonstrate the impact of our plant separates in hindering warmth prompted proteins. The calming action of plant extricates are given Figure 1.

Figure 1. The anti-inflammatory activity of Morinda citrifolia using aqueous and hydro ethanolic extract.equationequation.


The Morinda citrifolia contain the increased anti inflammatory activity in the hydroethanolic extact compared than aqueous. Its used to reduce the inflammatory disorders. The inflammatory response is a dynamic process involving complex interactions among inflammatory molecules that arise in any tissue in response to traumatic, infectious, toxic, or auto-immune injury. M. citrifolia leaves have a therapeutic potential that supports its use in traditional medicine, and could be developed as a commercial anti-inflammatory. Phytochemical constituents in Morinda citrifolia L have the potential to act as a source of useful drugs and also to improve the health status of the consumers as a result of the presence of various compounds that are vital for good health. Morinda citrifolia L is found to be a potent health vitalizer to the mankind.

Conflict of Interest



Get the App