A Preliminary study on phytochemical analysis of leaves of Bixa orellana (Kumkum)

This Article is published here.

Sanjeet kumar, M. Kabi and M. Kumari ,2010 “Study on phytochemical analysis from leaves of Bixa orellana (Kumkum)”, Emerging Science,Vol 2, No. 5 ,October 2010

Sanjeet Kumar

sanjeet.biotech@gmail.com

Abstract

In this study, the medicinal properties of Bixa orellana were evaluated by phytochemical screening of acetone & methanol extract .In acetone extract steroid, terpenoid & glycoside shown but in methanol extract tannin & flavonoid shown , these bioactive compounds indicated the medicinal value of  leaf of Bixa orellana.

Key words : Bixa orellana, Phytochemical screening, Medicinal value.

 Introduction



Fig : Plant of Bixa orellana



The state of Orissa possesses a rich bio-diversity of medicinal plants which are the richest and can provide diverse flora and natural vegetations, many of which are used in traditional medicine to cure various sickness and diseases. They not only serve as source of medicinal purpose, but also used for other purposes; which many due to their odoriferous nature and useful in flavoring or as food additives and preservatives.

                          The use of plants whether herbs, shrubs or trees, in parts or in whole, in the treatment and management of diseases and disorder date back to pre-historic days. Plant extracts or plant-derived compounds are likely to provide a valuable source of new medicinal compounds (Carvalho and Ferreira, 2001; Kayser and Kiderlen, 2001) and the urgent need for alternative treatment has led to screen natural products for potential therapeutic use in microbial and fungal infections. Plant parts have been used in folk medicine practices for the treatment of various ailments since antiquity. The medicinal properties of various plant materials and extracts have been recognized since the beginning of developing golden period of India. A rich store house of medicinal plants exist everywhere especially in Orissa which offers a vast reservoir of medicinal plants that have been categorized.

The current study is aimed to investigate the phytochemical properties of Bixa Orellana.

            Bixa orellana commonly called as Annato tree belongs to family Bixaceae and has been named as:-

·         Kumkum in Oriya

·         Sundri in Marathi

·         Latkan in Hindi & Bengoli.

·         Japhora in Tamil & Telgu.

·         Annato and Uruco in Brazil.

·         Achiott in Amezen India.

Botany:-

Shrub or small tree leaves ovate, base subcordate, apex acute to acuminate, Flowers, Brown, caduceus, sepals purplish red, broadly ovate to orbicular, petals purplish- pink to nearly white. Capsules ovoid, dark red, seed smooth.

Distributions:-

Cultivated and naturalized throughout the area and more in the central districts in Orissa. It is native of tropical America, now widely cultivated in tropical countries and frequently naturalized.

                    The outer coat of seed of Kumkum is used for colouring of fabrics in rural areas of Orissa; also seeds are very important for the isolation of active compounds which is Bixin, recognized as a safe colourent for different types of food like in butter, chocolate etc.

Traditional uses: -

Leaves are used naturally in Constipation, fevers, heartburn, asthma, gastrointestinal tonic (Lorenzi and Matos, 2002) Annatto extract is a natural food color, which is obtained from the outer coatings of the seeds of the Annatto tree (Bixa orellana L.) (Hagiwara et al., 2003). The use of annatto by the New World man dates back to ancient times. The indigenous Amazon people have used “urucum” for body. Painting for centuries. Seeds and leaves of the annatto tree were used by the Aztecs to prepare remedies for a variety of illnesses such as tonsillitis, asthma, pleurisy, rectal disorders, headache, jaundice, sunstroke, and burns (De Oliveira et al., 2003). Although the plant species originated in northern South America, it is now widely cultivated in tropical areas for commercial production (Hagiwara et al., 2003). Aqueous annatto extract had a hypocholesterolemic effect. (Heberth de Paula et al., 2009).

Chemical Constituents

Bixa orellana exhibited some level of antifungal activity when a group of Latin American plants were subjected to screening for antifungal activity (Freixma et al. 1998). In another experiment, the lipid fraction of B.orellana seeds was extracted using n-hexane and isolated by thin layer chromatography; it seems that Bixa seeds contain a higher concentration of delta-tocotrienol than any other vegetable species (Frega, Mozzon & Bocci 1998). Recently, three apocartenoids were isolated from annatto (B.orellana) and synthesized. They were: methyl (9Z)-8’-oxo-6, 8’diapocarten-6-oate (2), methyl Z)-10’-oxo-6, 10’diapocaroten-6-oate (4), and methyl (9Z)-14’-oxo-6,14’-diapocaroten-6-oate (5) (Haberli & Pfander 1999). In another experiment, three minor cartenoids were isolated from the seed coat of B.orellana fruits by chromatographic methods and, for the first time, geranylgeraniol has been found esterifies with a carotenoid carboxylic acid (Mercandante, steck & Pfander 1999).

Active compounds
Bixin and norbixin are the main pigments contributing to the color of Annatto extracts. Extracts are obtained from the outer layer of the seeds the principal pigment in annatto extract is cis-bixin, which is contained in the resinous coating of the seed itself. The seeds contain a higher concentration of delta-tocotrienol than any other vegetable species (Frega, Mozzon & Bocci 1998).

Materials and Methods:-

The leaves of Kumkum were collected from Ekamra kanan in Bhubaneswar, Orissa. The analysis for detection of phytochemicals was done at Biotechnotrics Pvt. Ltd. Bhubneshwar.

 Preparation of Plants Extract

The leaves of the plant were thoroughly washed with tap water. Each leaf was then dried with the help of tissue paper. The leaves were cut into small pieces and were washed with distilled water. The pieces of leaves were dried and their initial weight was taken. The leaves were then kept in a hot air oven at 60 degree Celsius for 12 hours. The pieces of leaves were then powdered with the help of mortar and pestle. The extract was prepared by using the solvents of methanol and acetone. The crude extracts were kept in sample bottles and was stored in refrigerator for further phytochemical analysis.

Phytochemical analysis

Phytochemical Screening was carried out following the methods provided by Sofowora, 1982 for qualitative analysis of methanol and acetone extracts of Bixa Orellana.( Kumkum)

1.    Test for Tannins :-

700 µl of leaf extracts was mixed with 50 ml of distilled water. 1% of ferric chloride was added to it as a drop. A dark green solution indicates the presence of tannins.

2.    Test for Flavonoids :-

Few drops of leaf extract were dissolved in 10% NaOH. Then concentrated HCl was added to it. A yellow solution that turns colourless indicates the presence of Flavonoids.

3.    Test for Terpenoids :-

1 ml of plant extract was taken and to it 400 µl of chloroform was mixed. Then few drops of concentrated Sulphuric acid was carefully added to form a layer. A reddish brown colouration of the interface was formed to indicate positive results for the presence of terpenoids.

4.    Test for Steroids :-

100 µl of plant extracts was taken and to it 400 µl of aceatic anhydride was added. To it few drops of concentrated sulphuric acid was added drops by drops.The formation of a brown ring indicates the presence of Steroids.

5.    Test of Saponins :-

About 1 ml of plant extracts was taken and to it 1% ferric chloride was added. Frothing or appearance of creamy mass of small bubbles shows the presence of saponins.

6.    Test for Glycosides :-

100 µl of plant extract was taken and to it 400 µl of acetic anhydride was added in a cool medium. Then few drops of concentrated sulphuric acid was added to it. The change of colour from violet to blue and then to green indicates the presence of Glycosides.

7.    Test for amino acids :-

2 ml of leaf extract which was dissolved in protein extracting buffer. To it 500 µl ninhydrin reagent solutions was added. The formation of purple colour indicates the presence of amino acids.

Results

Table no.1:-

Initial weight	Final weight
21.93 gm	7.75 gm

Table no. 2

The moisture % and the solid % of Bixa orellana leaves are:-

Moisture %	Solid %
64.66%	35.34%

Table no. 3:- Methanol Extract

Test	Results
Tannin	+ve
Flavonoid	+ve
Terpenoid	-ve
Saponin	-ve

Table no.4:- Acetone Extract

Test	Result
Tannin	-ve
Steroid	+ve
Terpenoid	+ve
Glycoside	+ve

Conclusion

The medicinal properties of the leaves of Bixa orellana could be attributed to the presence of one or more of the detected phytochemicals. The methanol extract of leaf contains flavonoids which has antioxidant properties. The acetone extract show positive for steroids which has indicative of stimulant properties.. The methanol extract also revealed the presence of terpenoids and it helps in the biosynthesis of lipids and increase cholesterol and decreases blood sugar.However, the pharmacological actions of the plant cannot be ascertained by only the qualitative result of the phytochemical screening. Further research can exhibit the detail pharmacological characteristics potent to its medicinal value.

References

·         Carvalho, P.B., Ferreira, E.I., 2001. Leishmaniasis phytotherapy. Nature’s leadership against an ancient disease-review. Fitoterapia 72, 599–618.

·         De Oliveira AC, Silva IB, Manhaes-Rocha DA,Paumgartten FJ (2003) Induction of liver monooxygenases by annatto and bixin in female rats. Braz J Med Biol Res 36:113-118.

·         (Frega, Mozzon & Bocci 1998. Frega, N., M.Mozzon,& F.Bocci.1998. Identification and estimation of tocotrienols in the annatto lipid fraction by gas chromatography mass spectrometry. Journal of the American Oil Chemists Society. 75 (12): 1723-1727.

·         Freixa, B., R. Vila, L. Vargas, N. Lozano, T. Adzet & S. Canigueral.1998.
Screening for antifungal activity in nineteen Latin American plants. Phytotherapy Research.12 (6): 427-430.

·         H.Sexena and M.Brahaman, The Flora of Orissa, Vol-1

·         Heberth de Paula Effect of an Aqueous Extract of Annatto (Bixa orellana) Seeds on Lipid Profile and Biochemical Markers of Renaland Hepatic Function in Hipercholesterolemic Rats, Braz. Arch. Biol. Technol. v.52 n. 6: pp. 1373-1378, Nov/Dec 2009.

·         Haberli, A.& H. Pfander. 1999. Synthesis of bixin and three minor carotenoids from annato (Bixa orellana).
Helvetica Chemica Acta.82 (5): 696-706.

·         Hagiwara A, Imai N, Ichihara T, Sano M, Tamano S,Aoki H, Yasuhara K, Koda T, Nakamura M, Shirai T(2003) A thirteen-week oral toxicity study of annatto extract (norbixin), a natural food color extracted from the seed coat of annatto (Bixa orellana L.), in Sprague-Dawley rats. Food Chem Toxicol 41:1157-1164.

·         (Kayser, O., Kiderlen, A.F., 2001. In vitro leishmanicidal activity of naturally occurring chalcones. Phytotherapy Research 15, 148–152.

·         Lorenzi, H., Matos, F.J.A., 2002. Plantas Medicinais do Brasil. Nova Odessa, Instituto Plantarum, S˜ao Paulo, p. 544.

·         Mercadante, A.Z., Steck, H. and Pfander, H. (1999). Three minor carotenoids from annatto (Bixa orellana).

Sanjeet Kumar, Regional Institute of Education (NCERT) Bhubaneswar.

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