INTRODUCTION:

Traditional herbal drugs are the major source of health care for more than two thirds of the world’s population. The World Health Organization is strongly advocating for wider acceptance and use of traditional medicines including Ayurveda and Unani. This primarily stems from belief that herbal medicines are safe, inexpensive and have no adverse effects.¹

Murraya koenigii Spreng. belonging to family Rutaceae, commonly known as “Curry Patta” is a deciduous aromatic shrub, up to a height of 6 meters.² Murraya koenigii is genus of tree, native to tropical Asia from Himalaya foothill’s of India to Shrilanka eastward through Myanmar, Indonesia, Southern China and Hainan. In India it occurs in foothill of Himalaya, Assam, Sikkim, Kerala, Tamilnadu, Andhra Pradesh and Maharashtra.^3,4

Various parts of Murraya koenigii have been used in traditional or folk medicine for the treatment of rheumatism, traumatic injury, snake bite and it has been reported to have antioxidant, antidiabetic, antidysenteric^5,6, antioxidant⁷, anticancer, anti-inflammatory⁸, antifungal⁹ and immunomodulatory activity¹⁰.

Though various parts of Murraya koenigii have been highlighted for different ethnopharmacological properties, research study report on pharmacognostic studies of stem bark is lacking. In the present study an attempt has been made to evaluate the pharmacognostical characteristics of Murraya koenigii stem bark.

MATERIAL AND METHODS:

Plant material:

The stem bark of Murraya koenigii was collected from Moshi, Pune in the month of October and was authenticated by Dr. Jayanti, Botanical Survey of India, Yerwada, Pune.

Processing of plant material:

The fresh stem bark was shed dried and then powdered and stored in air-dried containers.¹¹

Pharmacognostic evaluation:

The fresh stem bark of Murraya koenigii was studied for external visible characters such as size, shape, surface, fracture and organoleptic properties. The microscopic studies were carried out using Nikon camera attached with microscope. The clear free hand sections of stem bark previously soaked in water were boiled in chloral hydrate to remove the coloring matter. Then sections were stained with phloroglucinol and concentrated hydrochloric acid. The selected section was mounted on clean glass slide using glycerine and covered with coverslip. The section was then viewed under 10X and 45X. Some of the sections were also stained with dilute Iodine for starch, with Sudan Red for cuticle, Rhuthenium Red for mucilaginous cells and Acetic Acid for calcium oxalate.

The powder of stem bark was subjected to powder microscopy. Powder was first cleared with chloral hydrate and then stained with staining reagents phloroglucinol and concentrated HCl and then viewed under 10X and 45X.

Physicochemical evaluation:

Physicochemical evaluation like total ash value, acid insoluble ash value, water soluble ash value, sulphated ash value, moisture content, foaming index, swelling index, foreign organic matter were determined.

Extractive values were determined by using different methods including hot extraction, cold maceration and successive extraction. All parameters were determined as per WHO guidelines.

Fluorescence analysis:

The powdered drug and different solvent extracts of stem bark were examined under short UV (254 nm), long UV (366 nm) and day light using different solvents.¹²

Phytochemical screening:

Preliminary phytochemical screening of various extracts obtained in successive solvent extraction was carried out with different reagents and solvents by performing various qualitative chemical tests.¹³

RESULTS AND DISCUSSION:

The present study reports the pharmacognostical characteristics of Murraya koenigii stem bark. This can be used to determine correct identity and purity of plant’s part and detection of adulteration as well. To ensure reproducible quality of herbal products, proper control of starting material is utmost essential. Thus in recent years there has been an emphasis in standardization of medicinal plants of therapeutic potential. Despite the modern techniques, identification and evaluation of plant drugs by pharmacognostical studies is still more reliable, accurate and inexpensive means. According to World Health Organization the macroscopic and microscopic description of a medicinal plant is the first step towards establishing its identity and purity and should be carried out before any tests are undertaken.¹⁴

Pharmacognostic evaluation:

Macroscopic or organoleptic evaluation is a technique of qualitative evaluation based on the study of morphological and sensory profile. Organoleptic characters of stem bark were observed and recorded in Table 1.

Table 1. Organoleptic characters of Murraya koenigii stem bark.

Sr. No.	Characters	Observations
1	Color	Dark brown (unpeeled), Creamish brown (peeled)
2	Odor	Aromatic
3	Taste	Characteristic
4	Shape	Quelled longitudinally
5	Size	0.5-1 cm thickness
6	Fracture	Splintery
7	Surface	Rough, hard, numerous creamish spots on outer surface

Transverse section of stem bark (Fig.1) shows few layers of tangentially elongated cork cells. Phellogen is two to three layers of thin walled cells without any cellular content. Phelloderm is of five to six rows, thin walled. Cortex contains several layers of thin walled tangentially elongated cells containing reddish brown matter. Circular, lignified phloem fibers are present. Tangentional rows of stone cells separated by medullary rays are present. Medullary rays are 2-3 celled wide towards the outer part. Numerous oil cells are also present.

Powder microscopy of stem bark shows thin walled, polygonal with yellowish brown matter cork cells (Fig.2). Medullary rays are two to three seriate, extended upto cortex cells, radially elongated (Fig.3). Oil vacuole appear yellowish (Fig.4). Numerous, large, stratified thick lignified wall and narrow lumen phloem fibers are present (Fig.5). Starch grains are simple (Fig.6).

Physicochemical evaluation:

Various physicochemical parameters such as total ash value, acid insoluble ash value, water soluble ash value, sulphated ash value, extractive values, moisture content, foaming index, swelling index, foreign organic matter were determined and are depicted in Table 2, 3 and Table 4.

Table 2. Physicochemical parameters of Murraya koenigii stem bark.

Sr. No.	Parameters	Values obtained
1	Total ash	8.5 % w/w
2	Acid insoluble ash	0.5 % w/w
3	Water soluble ash	6.5 % w/w
4	Sulphated ash	4 % w/w
5	Moisture content	14 % w/w
6	Foaming index	Less than 100
7	Swelling index	Nil
8	Foreign organic matter	0.4 % w/w

Table 3. Extractive values of Murraya koenigii stem bark.

Sr. No.	Parameters	Values obtained
1	Water soluble extractive value (cold maceration)	27.2 % w/w
2	Alcohol soluble extractive value (cold maceration)	21.6 % w/w
3	Water soluble extractive value (hot extraction)	19 % w/w

Table 4. Successive Solvent Extractive Values of Murraya koenigii stem bark.

Sr. No.	Solvents	Consistency and color	Extractive value (%w/w)
1	Petroleum ether	Semi solid, brown	4.73
2	Chloroform	Semi solid, brown	1.17
3	Acetone	Solid, brown	1.35
4	Ethanol (95%)	Semi solid, dark brown	4.26
5	Water	Solid, brown	3.66

Fluorescence analysis:

Fluorescence analysis of powdered drug and different solvent extracts of stem bark was done and the results were recorded in Table 5.

Phytochemical screening:

Phytochemical screening of various extracts obtained in successive solvent extraction was carried out and the results were recorded in Table 6.

Table 5. Fluorescence analysis of Murraya koenigii stem bark.

Sr.No.	Solvents	Long UV (366 nm)	Short UV (254 nm)	Day light
1	Powder + Conc. HCl	Brown black	Green	Yellow brown
2	Powder + Conc. H2So4	Black	Brown black	Brown black
3	Powder + 10%NaOH	Brown	Brown yellow	Brown yellow
4	Powder + Conc.HNO3	Brown black	Green yellow	Yellow brown
5	Powder + Ammonia solution	Green	Green yellow	Yellow
6	Powder + Acetone	Yellow brown	Yellow	Cream
7	Petroleum ether extract	Yellow brown	Dark yellow brown	Brown
8	Chloroform extract	Yellow brown	Dark yellow brown	Brown
9	Acetone extract	Black brown	Dark yellow brown	Black brown
10	Ethanol extract	Brown	Yellowish brown	Black brown
11	Water extract	Green brown	Dark green	Green Brown

Table 6. Phytochemical screening of Murraya koenigii stem bark.

Sr. No.	Phytoconstituents	Petroleum ether	Chloroform	Acetone	Ethanol	Water
1	Alkaloids	+	+	+	+	+
2	Carbohydrates	+	+	+	+	+
3	Glycosides	-	-	-	-	-
4	Tannins	-	-	+	+	+
5	Proteins	-	+	-	-	-
6	Saponins	-	+	-	-	+
7	Steroids	-	+	-	-	+
8	Flavonoids	-	-	-	+	+

+ = Present, - = Absent

Fig 1. It shows T.S. of Murraya koenigii stem bark.

Fig 2. Cork

Fig 3. Medullary rays

Fig 4. Oil vacuole

Fig 5. Phloem fibers

Fig 6. Starch grains

Fig.-2-6: Microscopic characters of Murraya koenigii stem bark powder.

CONCLUSION:

Such a pharmacognostic study of stem bark of Murraya koenigii is useful for standardizing crude drug and can be used to differentiate closely related species. Various physicochemical parameters were established which can be important in detecting adulteration and mishandling of the crude drug. Phytochemical analysis showed the presence of many important classes of phytoconstituents. This indicates that the plant can be useful for treating different diseases because the therapeutic activity of a plant is due to the presence of particular class of compounds. Such a detailed study would be decisive in performing standardization of the stem bark material, preparation of its monograph, isolation of phytoconstituents, performing further pre-clinical and clinical investigations and manufacturing of its formulations.

ACKNOWLEDGEMENT:

The authors are thankful to Prof. S. N. Dhole, Principal Modern College of Pharmacy (For Ladies) for providing facilities to carry out this work.

REFERENCES:

1. Aggarwal S.S., Paridhavi M. Herbal Drug Technology, University Press (India) Pvt. Ltd., Hyderabad, 2007; p. 326.

2. Kritikar K.R., Basu BD. Indian Medicinal Plants, International Book Distributors, Dehradun, I, 2004. p. 472-479.

3. Parrota J. A., In; Healing Plants of Peninsular India, C.A.S.I. Publication, U.S.A., 2001, 639.

4. Prajapati, N. D., Purohit, S. S., Sharma, A. K., Kumar, T., In; A andbook of Medicinal Plants, 1st, Edn., Agrobios India, 2003, 401.

5. Kong Y.C., Zhang H.T., Cheng K.F., Soejarto D.D., Kan W.S. Sources of the anti-implanation alkaloid in the genus Murraya. J Ethnopharmacol 1986, 15: 195-200.

6. Keasri A.N., Kesari S, Singh S.K., Gupta R.K., Watal G. Studies on the glycemic and lipidemic effect of Murraya koenigii in experimental animals. J Ethnopharmacol 2007, 112: 305-311.

7. Patel R.S., Rajorhia. Antioxidative role of curry (Murraya koenigii) and betel (Piper betel) leaves in ghee. J. Food Sci. Tech. 1979; 16 (4): 158-160.

8. P. Muthumani, S.Venkatraman, K.V. Ramsesh, R. Meera , P. Devi, B. Kameswari, B.Eswarapriya. Pharmacological studies of anticancer, anti inflammatory activities of Murraya koenigii (Linn) Spreng in experimental animals. Journal of Pharmaceutical Sciences and Research. Vol.1(3), 2009, 137-141

9. Tripathi R.N., Pandey D.K., Tripathi N.N., Dixit S.N. Antifungal activity of pollens of some higher plants. Ind. Phytopathol. 1982; 35(2): 346-348.

10. Shah A.S., Wadake A.S., Juvekar A.R. Immunomodulatory activity of methanolic extract of Murraya koenigii (L.) Spreng. leaves. Ind. J. Exp. Biol. 2004; 46(7): 505-509.

11. 12.Quality control methods for medicinal plant materials, WHO Guidelines, Geneva. 2002. AITBS Publishers and Distributors, Delhi, p. 10-46.

12. Chase C.R., Pratt R.; Fluorescence of powdered vegetables drugs with particular reference to development of a system of identification. J. Am. Pharma. Assoc. 38(1949,) page 324-31

13. Khandelwal K. R., Practical Pharmacognocy, Techniques and Experiments, Edition 12, Nirali Prakashan, 2008, Page: 149-153

14. Anonymous. Macroscopic and microscopic Examination: Quality Control Methods for Medicinal Plant Materials, WHO, Geneva; 1998.

Received on 13.07.2017 Modified on 06.09.2017

Res. J. Pharmacognosy and Phytochem. 2017; 9(3): 177-181.

DOI: 10.5958/0975-4385.2017.00033.4