PHARMACOGNOSTIC STUDY OF ERANTHEMUM NIGRUM ROOTHTML Full Text
PHARMACOGNOSTIC STUDY OF ERANTHEMUM NIGRUM ROOT
D. S. N. B. K. Prasanth * and A. Lakshmana Rao
Department of Pharmacognosy, V. V. Institute of Pharmaceutical Sciences, Gudlavalleru - 521356, Andhra Pradesh, India.
ABSTRACT: Objective: To analyze the pharmacognostic characteristics and physiochemical parameters of the root of Eranthemum nigrum (E. nigrum). Methods: Microscopic characters and powder analysis had been carried out with the help of a microscope. The physiochemical properties such as loss on drying, total ash value, acid insoluble ash value, water-soluble ash value, extractive values and fluorescence of E. nigrum had been performed. Results: The color, shape, size, odor, and surface characteristics were reported from the root and powdered root material of E. nigrum. Light microscope images of cross section and powdered root revealed the presence of lignified xylem fibers, xylem vessels, cork cells, and parenchyma cells. Phytochemical testing confirmed the presence of steroids, alkaloids, tannins, saponins, carbohydrates, glycosides, amino acids and proteins. Physicochemical parameters such as moisture content, ash value, extractive value and fluorescent behavior of root powder have also been established. Conclusion: The morphological, microscopical and physicochemical parameter results provided in this paper may be utilized as a basis for the preparation of a monograph on E. nigrum root.
Pharmacognostic, Eranthemum nigrum, Lignified xylem vessels, Phytochemical and Physicochemical analysis
INTRODUCTION: Medicinal plants are usually playing a significant part in traditional medicines intended for the therapy of various health issues. However, a crucial hurdle, which has impeded the promotion in the usage of alternative medications in the developed countries, is the lack of evidence of documentation and absence of stringent quality control measures. Additionally, there is a dependence on the data of all study meted out on traditional medicines by way of documentation. Keeping this issue, it is now quite necessary to generate assurance about the standardization of the plant as well as its parts to be used as a medication.
During the process of standardization, we can take advantage of various techniques and methodology to achieve our goal in a phase-wise approach, e.g. pharmacognostic and phytochemical studies. These techniques and methods are helpful in recognition and standardization of the plant material. Appropriate characterization and quality assurance of starting material is a crucial step to ensure the reproducible quality of herbal medicine to assist people to justify its safety and effectiveness.
Because of this reason, we have executed pharmacognostic studies of Eranthemum nigrum belongs to family Acanthaceae 1. This sort of research is not going to help in authentication but additionally ensures reproducibility of herbal goods in promoting 2.In the present study, we have been focusing our exploration on one of the commonly available plants in India, i.e., Eranthemum nigrum, belongs to family Acanthaceae. The family Acanthaceae consists of almost 4000 species of exotic plants. Various species of genus Eranthemum being utilized traditionally for extensive kinds of ethnomedicinal purposes. The genus Eranthemum, with around 138 species, some of the important species include E. austrosinensis, E. burmanicum, E. capense, E. ciliatum, E. erythrochilum, E. griffithii, E. macrophyllum, E. macrostachyus, E. obovatum, E. pulchellum, E. purpurascens, E. roseum, E. strictum, E. tapingense, E. tubiflorum and E. watti.
The Eranthemum nigrum (Acanthaceae) is native to the Pacific Islands. The shrub attains height of 1.5 - 1.8 m. The upper surface of leaves is blackish purple and the lower surface purplish with dark veins. The flowers are in erect terminal spikes, white and spotted rose at the base 3. Plants are adapted to partial shade. The leaves are elliptical, glossy or dull with smooth margins and acute tips 4, 5. All parts of this plant are widely used as a folklore medicine for the treatment of various ailments by the Indian traditional healer. Ethnomedicinally, the genus Eranthemum has been documented various pharmacological activities including antipyretic 6, antidiabetic 7, antiulcer 8, antimicrobial 9, larvicidal, ovicidal and pupicidal against Anopheles stephensi 10, gastroprotective 11 and anti-inflammatory 12.
A literature study and screening of scientific data says a lot of native medicines have already been investigated as regards their botany and chemistry is concerned; however a systematic standardization including pharmacognostical and physicochemical study is still lacking. The present investigation of Eranthemum nigrum. (Acanthaceae) is therefore taken up to establish certain botanical and chemical standards which would help in crude drug identification as well as in checking adulteration, if any. Further, the study will greatly help in quality assurance of finished products of herbal drugs 13, 14.
MATERIALS AND METHODS:
Plant Collection and Authentication: The plant was obtained from V.V. Institute of Pharmaceutical Sciences, Gudlavalleru, Krishna District of Andhra Pradesh, India during September 2017 and authenticated by Dr. K. Madhava Chetty, Taxonomist at Sri Venkateswara University, Tirupati, India. The plant was deposited at the herbarium for future reference. One portion of the root is preserved in formalin: acetic acid: alcohol mixture for histological studies and the remaining portion was shade dried, powdered and sieved through 20 mesh and kept in an airtight container for future use.
Chemicals: All analytical grade chemicals were utilized in this study were procured from E. Merck, Germany absolute alcohol, phloroglucinol, acetic acid, chloral hydrate, H2SO4, NaOH, HNO3, FeCl3, distilled water, conc. HCl and chloroform.
Morphological Evaluation: Organoleptic evaluation of E. nigrum root has been carried out by the color, size, odor, shape, and taste as per WHO quality control methods of herbal medicine 15.
Preparation of Sections: Microscopic studies had been done by preparing thin hand section of the root with the help of sharp cutting edge of the blade, then cleared with chloral hydrate solution, stained with phloroglucinol-hydrochloric acid (1:1) and mounted in glycerin.
Powdered Microscopy: The powder microscopy was carried out by the procedure described in Khandelwal 16.
Quantitative Analysis: The quantitative examinations including stomatal number, stomatal index, vein islet number, and vein termination number were studied using standard method 2.
Preparation of Extracts and Preliminary Phytochemical Analysis: The powdered material had been extracted with various solvents according to its polarity, i.e., chloroform, methanol, and water. 5 g root powder was extracted with 20 ml of the respective solvent by maceration at room temperature for 24 h. Then, filtered through Whatmann filter paper and collect the filtrate, concentrated with rota-evaporator. Then, the extracts had been subjected to preliminary phytochemical screening according to standard methods 16, 17.
Physicochemical Analysis: Physicochemical parameters such as ash value, moisture content and extractive values were determined according to the procedures mentioned in WHO quality control methods for herbal materials 18.
Fluorescence Analysis: Various reagents were utilized to check the fluorescence activity. In this, 0.1 g of root powder was blended with 1.5 ml of respective reagent Table 4. The mixture was placed on a slide for a minute and observed under visible light, short ultra-violet light (254 nm) and long ultraviolet light (365 nm) (19).
Morphological Characteristics: The morpho-logical characteristics of E. nigrum root were described in Fig. 1 and Table 1.
FIG. 1: MORPHOLOGICAL FEATURES OF ROOT OF ERANTHEMUM NIGRUM
TABLE 1: MORPHOLOGICAL CHARACTERISTICS OF ROOT OF ERANTHEMUM NIGRUM
Root: The transverse section of the root of E. nigrum showed the presence of cortex (Phellem) shows 3- 4 polygonal thick walled parenchymatous cells filled with brown content. Cork cambium (Phellogen) was made up of 3 - 5 layered narrow, tangentially elongated parenchymatous cells. Secondary cortex (Phelloderm) is 4 - 6 layered rows of tangentially elongated thin-walled cells. The endodermis showed the presence of phloem and xylem. The phloem is present in between the medullary rays. The medullary rays are parenchymatous and are uniseriate to triseriate, majorly biseriate.
Radially arranged vascular bundles were present in which, phloem is well developed and shows the presence of phloem fibers, which are non-lignified. It also showed the presence of phloem parenchyma. The xylem region was similar to the phloem region and was also surrounded by uniseriate to triseriate medullary rays. Xylem tissue consists of spiral xylem vessels, xylem fibers and xylem parenchyma Fig. 2.
Powder Microscopy: The crude powder of root was buff in color with characteristic odor and taste. Microscopic study of the powder showed revealed different characters such as diacytic stomata, covering trichomes, xylem vessels and parenchyma cells Fig. 6.
FIG. 6: POWDER MICROSCOPY OF ROOT OF ERANTHEMUM NIGRUM. (a) Parenchyma cells (b) lignified xylem vessels (c) Cork cells (d) Medullary rays with xylem vessels.
Preliminary Phytochemical Analysis: The results of the qualitative phytochemical analysis of crude powder of E. nigrum root are shown in Table 2.
Physicochemical Parameters: The results attained from various determinations of physicochemical analysis are produced in Table 3.
TABLE 2: PRELIMINARY PHYTOCHEMICAL ANALYSIS OF ERANTHEMUM NIGRUM ROOT
|Phytoconstituents||Method||Aqueous Extract||Methanolic Extract||Chloroform Extract||Pet. ether
|Zn. hydrochloride test||+||+||-||-|
|Lead acetate test||+||+||-||-|
|Volatile oil||Stain test||-||-||-||-|
|Potassium dichromate test||-||-||-||-|
|Steroids and Triterpenoids||Salkowski test||+||+||-||+|
|Acid compounds||Litmus test||-||-||-||-|
|Amino acids||Ninhydrin test||+||+||-||-|
“+” -Present; “–“-Absent
TABLE 3: PHYSICOCHEMICAL PARAMETERS OF ROOT POWDER OF ERANTHEMUM NIGRUM
|Parameters||Values % w/w|
|Moisture content (Loss on drying)||8.25 ± 0.63|
|Total ash||6.56 ± 0.23|
|Acid-insoluble ash||4.12 ± 1.22|
|Petroleum ether soluble extractive value||0.63 ± 0.05|
|Chloroform soluble extractive value||2.12 ± 0.06|
|Ethyl acetate soluble extractive value||6.24 ± 0.05|
|Alcohol soluble extractive value||8.66 ± 0.25|
|Water soluble extractive value||10.12 ± 0.22|
Fluorescence Analysis: Fluorescence analysis of root powder was performed out after treating with different solvents. Fluorescence was observed at 254 and 365 nm comparing its change of color in the visible light. The observations are presented in Table 4 shows the variation in color.
TABLE 4: FLUORESCENCE ANALYSIS OF ERANTHEMUM NIGRUM ROOT POWDER
|At short (254 nm)||At long (365 nm)|
|FeCl3||Brownish yellow||Dark blue||Black|
|Picric acid||Brownish yellow||Dark blue||Black|
|Ethyl acetate||Black||Black||Greenish black|
DISCUSSION: Indian systems of medicine utilize the majority of the crude drugs which are of plant origin. It is important that standards need to be set down to control and check the identity of the plant and confirm its quality before use. Hence a detailed pharmacognostic assessment is extremely an important prerequisite. In accordance with World health organization (WHO) the organoleptic and histological description of a medicinal plant could be the first step towards establishing its identity and purity and should be performed before to any tests tend to be undertaken 20. E. nigrum, extensively utilized in conventional medicines has tremendous therapeutical potential due to its various biological activities. The prominent diagnostic characteristics of root were parenchyma cells, lignified xylem vessels, cork cells, medullary rays with xylem vessels. These characters can be utilized for standardization of drugs as well as useful for the preparation of plant monograph and also reduces the possibilities of adulteration, when the drug is available in the powdered form studies of physicochemical parameters can serve as an important source to judge the purity and quality of crude drugs.
Ash values are utilized to establish the quality and purity of the crude drug. It implies the existence of various impurities like carbonate, oxalate, and silicate. The water-soluble ash is water-soluble part of total ash, employed to calculate the number of inorganic substances found in the drugs. The insoluble acid ash comprises mostly silica and indicates contamination with earthy matter. The moisture content of drugs might be at a minimum level to suppress the growth of microorganisms like bacteria, yeast or fungi during storage. The extractive values are helpful to judge the chemical constituents present in the crude drug and also assist in the evaluation of particular constituents soluble in a specific solvent. Total ash and acid insoluble ash is essential indices to illustrate the quality and purity of the herbal medicine.
Total ash consists of physiological ash, which is derived from plant tissue itself, and nonphysiological ash that is usually from atmosphere contaminations includes sand and soil. Total ash content alone is not adequate to indicate the quality of herbal medicine because the plant materials usually contain a significant level of physiological ash, calcium oxalate in particular.
Therefore, the acid insoluble ash content is another index to indicate the quality of herbal medicine 20, 21 22. The phytochemical analysis of extracts viz., petroleum ether, chloroform, methanol, and water were analyzed and it indicates the presence of steroids, alkaloids, tannins, saponins, carbohydrates, glycosides, amino acids and proteins.
CONCLUSION: Standardization of herbal drugs is very much crucial because they are produced from heterogeneous sources which could result in variations. These kinds of variations can cause spurious results in various pharmacological and phytochemical studies. Eranthemum nigrum root is recognized for many therapeutical properties; therefore, the current study might be beneficial to supplement the information in respect to its identification, authentication, and standardization; no such information is available for the same till date.
CONFLICT OF INTEREST: Nil
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How to cite this article:
Prasanth DSNBK and Rao AL: Pharmacognostic study of Eranthemum nigrum root: Int J Pharmacognosy 2018; 5(4): 242-48. doi link: http://dx.doi.org/ 10.13040/ IJPSR.0975-8232.IJP.5(4).242-48.
This Journal licensed under a Creative Commons Attribution-Non-commercial-Share Alike 3.0 Unported License.
D. S. N. B. K. Prasanth* and A. L. Rao
Department of Pharmacognosy, V.V. Institute of Pharmaceutical Sciences, Gudlavalleru, Andhra Pradesh, India.
24 November 2017
05 January 2018
13 February 2018
01 April 2018