PHARMACOGNOSTICAL INVESTIGATION AND TOTAL PHENOLIC CONTENT OF DALBERGIA LATIFOLIA (ROXB) BARK

PHARMACOGNOSTICAL INVESTIGATION AND TOTAL PHENOLIC CONTENT OF DALBERGIA LATIFOLIA (ROXB.) BARK

M. Khalid ^*, J. Akhtar, Badruddeen, M. Arif and Kuldeep Singh

Faculty of Pharmacy Integral University, Dasauli Kursi Road, Lucknow - 226026, Uttar Pradesh, India.

ABSTRACT: Dalbergia latifolia commonly known as bilayatti shisham (Fabaceae) is a medicinal plant. The dry bark is brown colored astringent characters. Traditionally various species are reported to be used as aphrodisiac, abortifacient, expectorant, anthelmintic, antipyretic, appetizer, allays thirst, vomiting, burning sensation, cure skin diseases, ulcers Ayurvedic practice, bark is used as leucoderma, leprosy edema and bladder disorder. The bark minerals, acids, phenolic compounds, flavonoids, have characteristic smell astringent taste. This study deals with the pharmacognostical evaluation of the dried bark of Dalbergia latifolia which includes macro and microscopic studies, determination of physicochemical.

Dalbergia latifolia, Physicochemical parameter, Phenolic and flavonoid content

INTRODUCTION: The genus consists of 300 species and about 25 species occur in India. Many species of Dalbergia are important timber trees, valued for their decorative and often fragrant wood, rich in aromatic oils ^{1, 2}.

Traditionally various species are reported to be used as aphrodisiac, abortifacient, expectorant, anthelmintic, antipyretic, allays thirst, vomiting, burning sensation, cures skin diseases, ulcers, diseases of the blood, reduces obesity, used in leucoderma, dyspepsia, dysentery, for diseases of the eye and nose, syphilis, stomach troubles, leprosy, leucoderma, scabies and ringworm ^{3, 4, 5}. The present paper is a compilation of the phytoconstituents that have been identified in this genus and the traditional and reported biological activities.

Some phytoconstituents namely flavonoids, isoflavonoids, glycosides, steroids, etc.; have been isolated from the various species of the genus.  Dalbergia latifolia (Roxb.) Family- Fabaceae ⁵, a larg glabrous tree a single stem with characteristic smells ⁶. The tree has grey bark that peels in long fibers, compound leaves and bunches of small flowers ⁷. The bark is grey, thin with irregular short cracks, exfoliating in fibrous longitudinal flakes ⁸. It is distributed in Bihar, Bundelkhand and Central India ⁹.

It contains dalbinol a new 12a-hydroxyrotenoid ¹⁰, sisafolin coumarin from seeds, β-sitosterol, also contain dalbergichromene, lupeol, latifolin, and dalbergin from the bark of the tree, heartwood contains latinone, neoflavonoid dalcriodon ¹¹, ¹² and latinone, a substituted phenanthrene-1, 4-quinone was isolated from Dalbergia latifolia ¹³. Ethnomedicinally, the stem bark contains tannin is used for the treatment of leprosy, obesity, and worm. The genus consists of 300 species, and about 25 species occur in India. Many species of Dalbergia are important timber trees, valued for their decorative and often fragrant wood, rich in aromatic oils ¹⁴.

Traditionally various species are reported to be used as aphrodisiac, abortifacient, expectorant, anthelmintic, antipyretic, appetizer, allays thirst, vomiting, burning sensation, cures skin diseases, ulcers, diseases of the blood, reduces obesity, used in leucoderma, dyspepsia, dysentery, for diseases of the eye and nose, syphilis, stomach troubles, leprosy, leucoderma, scabies and ringworm ⁹, ¹⁵.

MATERIAL AND METHODS:

Materials: The bark material was collected in October from the Delhi market and identified from the Hamdard University New Delhi. Folin-Ciocalteus’s phenol reagent and sodium carbonate were from Merck chemical supplies (Darmstadt, Germany), routine and gallic acid purchase from Qualigens chemical suppliers.

Methods:

Microscopy: Transverse section (TS) of the root was cut by free hand sectioning and stained with different safranin and aniline blue. The various histological parts examined and drawn with the help of camera lucida ¹⁶ were performed for the presence of lignin, suberin, tannins, mucilage, starch grains and types of crystal present. Histochemical color reactions of the powdered drug were carried out with ruthenium red, iodine solution, Millon’s reagent, and Dragendorff’s reagent for the detection of mucilage, starch, protein, and alkaloids respectively ¹⁷.

Physico-Chemical and Fluorescence Analysis: Loss on drying, total ash, insoluble acid ash, water-soluble ash, and crude fibers contents was performed as per Indian Pharmacopoeia ¹⁸. The extract of the powdered fruit was prepared with different polar and non-polar solvents for the study of successive extractive values. Fluorescence analysis of the powder drug was carried out with different chemical reagents in the day (254 nm) and UV light (365 nm). The dry powder drug was studied on glass slide whereas the different extracts were studied by adsorbing the extracts on Whatmann filter paper ¹⁹.

Quantitative Estimation: For the quantitative estimation, 100 g of the powdered drug was successively extracted in a Soxhlet apparatus with various solvents like petroleum ether, chloroform, ethyl acetate, methanol and water ²⁰. The extracts were dried on a water bath, weighed and the color of the extracts was also observed. The different extracts were subjected to qualitative estimation for the presence of various phytoconstituents ²¹.

Determination of Total Phenolic Content: A total phenolic content in the B. diffusa extract was determined by the modified Folin-Ciocalteu method ²². An aliquot of the extracts was mixed with 5 ml Folin-Ciocalteu reagent (previously diluted with water 1:10 v/v) and 4 ml (75 g/l) of sodium carbonate. The mixtures were allowed to stand for 30 min at 40 °C for color development. Reagent blank using distilled water was prepared. The total phenolic content was calculated with the help of calibration curve prepared by repeating the operation using 1 ml of gallic acid solutions at concentrations (50,100, 150, 200, 250, 300 μg/ml) in distilled water.

Determination of Total Flavonoid Content: Total flavonoid content of B. diffusa was estimated by colorimetric method ²³. The extract was added in a volumetric flask (1 ml containing 10 mg/ml) of each followed by distilling water. The extract was mixed with 5% solution of sodium nitrite. After 5 min 0.3 ml of 10% AlCl₃ and after 6 min 2 ml of 1M-NaOH was added. Made up the volume to 10 ml with distilled water and the mixture of the volumetric flask were mixed thoroughly. The Absorbance of the mixture was measured at 510 nm against blank. The total flavonoid content was calculated with the help of the calibration curve and prepared standard rutin solutions at concentrations (50, 100, 200, 300, 400, 500 μg/ml) in distilled water.

RESULTS AND DISCUSSION:

Macroscopic & Microscopic Characters: The surface of the bark is rough, flat; fibrous, reddish brown in color.

FIG. 1: DALBERGIA LATIFOLIA BARK

They have longitudinal fissures. It is fibrous in texture, and the cut surface is smooth with a slight odor. The bark has 2 cm thick, 10-15 inch wide and 5-10 cm long Fig. 1.

It is differentiated into outer cork cell and inner cortex. The outer bark consists of simple stratified consist of 4-7 layers of rectangular cork cells. The cells are uniform tangentially oblong, and walls are suberized. The phelloderm is broad and prominent consisting of seven to ten layers in radial cells. The phelloderm cells are thin-walled with collapsed phloem zone is the abundance of prismatic calcium oxalate crystals disturbed in the parenchymatous cell Fig. 2.

FIG. 2: TRANSVERSE SECTION DALBERGIA LATIFOLIA BARK

Analysis of the Powder Bark: The powdered bark reveals fibrous light brown with slight odor and mucilaginous astringent taste. Simple hexagonal types of cork cells are found. Cork cells are stratified, appears like benzene ring in surface view.

FIG. 3: POWDER MICROSCOPY OF DALBERGIA LATIFOLIA BARK

Stone cells are containing with parenchymatous cells in the groups. Phloem fibers with linear fusiform arrangement are visible as highly lignified and pitted, pinkish colored, Calcium oxalate crystals are also present in prisms shaped scattered all over the powder. Prominent modularly rays are also observed in tangential longitudinal view. Black blue color starch grains are also seen inside the cells that are scattered over the slide observed when treated with iodine ²⁴ Fig. 3.

Histochemical Color Reactions: The powdered drug was pressing between filter paper mechanically no greasy stains were observed indicating the absence of fatty oil. When the powdered drug was mixed with water in a test tube and shake well frothing was not observed indicating saponins was absent. Powdered bark was pass through 60 mesh and mounted with different chemical reagents ruthenium red solution, Dragendorff reagent, conc. NaOH, anisaldehyde, chloral hydrate, iodine, and phloroglucinol + HCl were used for detection of the color of the powdered drug respectively Table 1.

TABLE 1: HISTOCHEMICAL COLOUR REACTION OF POWDER DRUG OF DALBERGIA LATIFOLIA

Physicochemical and Fluorescence Analyses: Physicochemical analyses of a powdered drug like a loss on drying, ash values, crude fibers and successive extractive values with different solvents of powdered root were analyzed. The percentage of all values in triplicate and their mean values ± SEM were calculated concerning the air-dried drug Table 2.

The changes in the color of D. latifolia bark powder under UV radiation about daylight were observed with different chemical reagents; it showed different colors reaction of powder indicating the presence or absence of chemical constituents Table 3. The fluorescence analyses of powdered drug play a vital role in the determination of quality and purity of the drug.

TABLE 2: QUANTITATIVE STANDARDS FOR THE DALBERGIA LATIFOLIA BARK

TABLE 3: FLUORESCENCE ANALYSIS OF POWDER DALBERGIA LATIFOLIA BARK

Extractive Values: The qualitative phytochemical analysis of D. latifolia bark extract was found to be 0.75 ± 0.14, 0.18 ± 0.02, 1.13 ± 0.03, 5.58 ± 0.14 and 5.56 ± 0.15 in petroleum, chloroform, ethyl acetate, alcohol, and aqueous extract respectively. All values in triplicate and their mean values ± SEM were calculated concerning the air-dried drug Table 4.

TABLE 4: EXTRACTIVE VALUES FOR CRUDE BARK OF DALBERGIA LATIFOLIA

 Qualitative Analysis: The presence or absence of different phytoconstituents viz. carbohydrate, glycoside, alkaloids, protein, tannins, flavonoids and terpenoids were detected by the phytochemical screening methods with different chemical reagents ¹⁷. Ethanolic and water extracts of the roots powder showed positive results for carbohydrate, glycoside, alkaloids, protein, tannins, saponins, flavonoids and terpenoids. The chloroform and ethyl acetate extract show positive results for terpenoids. Petroleum ether extracts have resinous matter which was not dissolved in other solvents Table 5.

Determination of Total Phenolics and Flavonoids Contents:  Table 6 indicated that the total phenolic and flavonoids content in D. latifolia bark of ethanolic extract had a higher level of phenolic compounds. The maximum absorbance of 0.73 was observed at a concentration of 210±1.51 µg/ml of extract. Gallic acid used as a standard which gave a maximum absorbance of 0.92 nm was observed at a concentration of 100 µg/ml and flavonoid observed in this plant 46 ± 3.61 µg/g which is equivalent to 175 µg/ml of rutin standard Table 6.

TABLE 5: QUALITATIVE CHEMICAL ANALYSIS OF DALBERGIA LATIFOLIA BARK EXTRACT

(+)  Present,    (-) Absent

The antioxidant activity mainly due to the redox properties ²⁵, which showed an important activity in adsorbing and neutralizing free radicals, entrapments of singlet and triplet oxygen, or oxidizing peroxides. The results of this study suggested that phenolics are important components of these plants.

TABLE 6: POLYPHENOL CONTENT OF DALBERGIA LATIFOLIA BARK EXTRACT

CONCLUSION: The present study of the bark powdered indicated the presence of carbohydrate, glycoside, alkaloid, protein, tannin, flavonoid and terpenoid. Pharmacognostical studied of the plant, in this study it was found that the D. latifolia bark had minerals, organic acids, flavonoids and phenolic compounds which has to found possesses antioxidant, mast cells stabilizing effects. The constituents of this plant have a tremendous impact on the health care system and may provide medical health benefits including the prevention and or treatment of diseases. Polyphenols traditionally have been considered to possess an anti-nutrient effect.

Recent interest in food phenolics has also increased greatly owing to their antioxidant capacity and possible beneficial implication in human health such as in treatment and prevention of cancer, cardiovascular diseases, and other pathological conditions.  The powder drug was tested for the presence of various inorganic elements such as potassium, magnesium, phosphorus, iodine respectively which have good neutraceuticals potentiality and can be used as a food supplement, preventive medicine provide medical health benefits including the prevention or treatment of diseases.

ACKNOWLEDGEMENT: Nil

CONFLICT OF INTEREST: Nil

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    How to cite this article:

    Khalid M, Akhtar J, Badruddeen, Arif M and Singh K: Pharmacognostical investigation and total phenolic content of Dalbergia latifolia (Roxb.) bark. Int J Pharmacognosy 2015; 2(5): 248-53. doi link: http://dx.doi.org/10.13040/IJPSR.0975-8232.IJP.2(5).248-53.

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