PHARMACOGNOSTIC AND PHYTOCHEMICAL EVALUATION OF NYCTANTHES ARBOR-TRISTIS STEMHTML Full Text
PHARMACOGNOSTIC AND PHYTOCHEMICAL EVALUATION OF NYCTANTHES ARBOR-TRISTIS STEM
Bhuwan Chandra Joshi * 1, Nilesh Chauhan 1, Sukanya 2 and Sushmita Uniyal 3
Department of Pharmacognosy 1, Sardar Bhagwan Singh Post Graduate Institute of Biomedical Sciences and Research, Balawala, Dehradun - 248001, Uttarakhand, India.
Department of Pharmacy 2, Central University of Rajasthan Bandarsindri, Kishangarh - 305817, Rajasthan, India.
Gyani Inder Singh Institute of Professional Studies 3, Dehradun - 248003, Uttarakhand, India.
ABSTRACT: Objective: To study detailed pharmacognostic characters of the stem of Nyctanthes arbor-tristis (Oleaceae), along with their physicochemical parameters, fluorescence analysis, and phytochemical screening. Methods: The pharmacognostic characters were determined in terms of macroscopy, microscopy, powder microscopy, physicochemical analysis, fluorescence analysis and preliminary phytochemical investigation of the plant stem. Results: The microscopic study shows the general characteristic of the stem. Physico-chemical investigation shows the total ash, acid insoluble ash, water-soluble ash were 8.69 ± 0.17% w/w, 0.21 ± 0.11% w/w, and 3.92 ± 0.05% w/w respectively. Phytochemical analysis revealed the presence of various phytochemical groups like alkaloids, glycosides, steroids, phenolic, tannins constituents. Conclusion: It can be concluded that the established pharmacognostic profile of Nyctanthes arbor-tristis stem will help develop pharmacopoeial standards for correct identification and quality control.
Nyctanthes arbor-tristis, Pharmacognosy, Fluorescence analysis
INTRODUCTION: India has a rich heritage of traditional medicine and traditional health care systems that have been flourishing for many centuries. Nowadays, the prevalent use of traditional medicines in developed countries is trending, and it has become more popular throughout the world. Traditionally, important large shrub of tropical and subtropical regions of the world have been used to counteract disease 1.
Nyctanthes arbour-tristis L. (Oleaceae) commonly known as Parijat or Harsinghar, a large hardy shrub or small tree up to 5-10 m in height, widely occurring in outer Himalayan ranges from Kashmir to Nepal and throughout India up to 1000-1500 m altitude 2.
The name ‘Nyctanthes’ has been coined from two Greek words ‘Nykhta’ means night and ‘anthos’ means flower. It is also planted in gardens due to its highly fragrant flowers 3. It is a shrub or small tree, with drooping branches and quadrangular branchlets. Leaves are opposite, ovate, acute or acuminate, entire slight cuneate. Flowers are small, 3-7 in the head, arranged in trichotomous cymes, delightfully fragrant, sessile, slender, and hairy; corolla glabrous, orange colored and lobes are white 4, 5. Fruits are capsules of 1-2 m in diameter, long and broad, compressed, 2 celled separating into 2 flat one-seeded carpels, reticular veined and glabrous 6. Different parts of this plant are used in folk-medicines 7, 8. The leaves are bitter, useful in chronic fever 9, malarial fever 10, obstinate sciatica, constipation, hemorrhoids 11 and eczema 12. The flowers are astringent, stomachic, and useful in dyspepsia, greyness of hair and baldness 11. The plant elaborates different classes of organic compounds of medicinal importance including alkaloids, terpenes, steroids, β-sitosterol, glycosides, iridoid glycosides, arbortristoside-A, B, C, D, E 13, 14, 15, 16, 17, 18.
Different parts of this plant are used in Indian systems of medicine for various pharmacological actions like as anti-leishmaniasis, anti-viral, anti-fungal, antibacterial, anti-pyretic, antihistaminic, anti-malarial, antioxidant, hepatoprotective, and anti-inflammatory activities19. The literature survey and scientific data revealed that no systematic pharmacognostical parameter had been carried out on the stem of Nyctanthes arbor-tristis Linn. till date. Hence, the objective of the present study is to evaluate various pharmacognostic parameters such as macroscopy, microscopy, physicochemical and phytochemical evaluations of the Nyctanthes arbor-tristis Linn. (Stem).
MATERIALS AND METHODS:
Chemicals and Instruments: Phloroglucinol, glycerin, hydrochloric acid, potassium hydroxide and all other chemicals used in the study were of analytical grade.
Plant Material: The stem of Nyctanthes arbor-tristis was collected from Dehradun, Uttarakhand, India in August 2016 and authenticated by Dr. S. K. Srivastava, Botanical Survey of India, Northern Regional Center, Dehradun, where a voucher specimen (specimen no. 116216) has been deposited.
Macroscopic and Microscopic Analysis: Macroscopic studies were done using a simple microscope. The color, shape, size, taste, and odor of the stem were determined. Microscopic study of the fresh stem was carried out by preparing a thin transverse section and staining it with concentrated hydrochloric acid: phloroglucinol (1:1). Photographs sections were carefully taken. The dried stem was powdered and treated with 5% KOH solution followed by staining with concentrated hydrochloric acid - phloroglucinol (1:1) for 5 min and mounted in 50% glycerine solution 20, 21, 22.
Physiochemical Analysis: Physiochemical parameter such as ash values (total ash, acid insoluble ash) and extractive values (water soluble, alcohol soluble extractives) were determined using the powdered drug. The moisture content was detected by loss on drying method 23, 24, 25.
Fluorescence Analysis: For the fluorescence analysis of stem powder it was treated with various chemicals and was observed exclusively to different wavelengths of ultraviolet (254 nm and 365 nm) and visible light for observing characteristic color presentation 22, 26, 27.
Pre-liminary Phytochemical Screening: Pre-liminary phytochemical screening was qualitatively tested for the presence of phytochemicals as per described standard methods 22, 28, 29, 30.
Macroscopic Characteristics: Organoleptic characters of stem depicted that the stem was woody having light grey to greenish in color with characteristic odor and taste. The stem was 1-10 m tall, erect and branched.
Stem Microscopy: Section of stem appears quadrangular and revealed the following tissues: The epidermis was single, layered consisting of rectangular cells with a thick continuous cuticle on the epidermis along with many multicellular hairs. Cortex was several cells deep below the four protruded comers while only a few layers deep at the other places just beneath the epidermis. It was differentiated into collenchyma and parenchyma. Many intercellular spaces were present, and the region extended up to the vascular tissue. Vascular bundles were present in the cortex; each protrudes bulb containing one. The pointed xylem end was faced towards the outer side in each of the conical bundle. In other words, the vascular bundle was conjoint, collateral, open an exarch.
The microscopy revealed that the endodermis was not well developed. The pericycle was observed in the form of sclerenchymatous patches. The vascular system was composed of primary phloem, secondary phloem, cambium, primary xylem, and secondary xylem. Crushed primary phloem was irregularly present in patches below the pericycle. The secondary phloem consisted of sieve tubes, companion cells, phloem parenchyma and was present in the form of a continuous ring. Cambium was present as one to three cells thick continuous layer in between phloem and xylem. Secondary xylem was present just inner to the cambial ring and consisted of mainly thick walled woody parenchyma and fibers. Tracheids and vessels were also observed. Primary xylem was situated just near the pith in a way facing its protoxylem towards the center. Pith was found to be thin-walled and parenchymatous as shown in Fig. 1.
Stem Powder Microscopy: Microscopic observation of Nyctanthes arbor-tristis stem indicated the presence of parenchyma cells, collenchyma cells, fiber, xylem vessels, and calcium oxalate crystal as shown in Fig. 2.
FIG. 1: TRANSVERSE SECTION OF STEM OF NYCTANTHES ARBOR-TRISTIS VIEWED AT 10x; A- WITHOUT STAINING REAGENT, B- WITH STAINING REAGENT
FIG. 2: POWDERED CHARACTERISTICS OF THE STEM PARTS OF NYCTANTHES ARBOR-TRISTIS SHOWING A: COLLENCHYMA CELL (CC) B: PARENCHYMA CELLS (PC) C: CALCIUM OXALATE CRYSTALS (CO) D: XYLEM VESSELS (XV) E: FIBER
Physicochemical Parameters: Ash value of the drug gives an idea about the earthy matter or inorganic composition and other impurities present along with the drug. Various physicochemical parameters such as total ash, acid insoluble ash, and water soluble ash of Nyctanthes arbor-tristis stem were found to be 8.69 ± 0.17, 0.21 ± 0.11 and 3.92 ± 0.05% w/w, respectively. However, 15.93 ± 0.46% w/w alcohol soluble and 18.31 ± 0.46% w/w water soluble extractives were observed. The moisture content of stem powder was nearly 5.49 ± 0.02% w/w Table 1.
Fluorescence Analysis: Fluorescence analysis of stem powder was carried out after treating it with several solvents and chemicals. Fluorescence was observed at 254 and 365 nm comparing its change of color in visible light. The observations are presented in Table 2.
Pre-liminary Phytochemical Screening: Pre-liminary phytochemical screening of Nyctanthes arbor-tristis is shown in Table 3.
TABLE 1: PHYSICOCHEMICAL CONSTANTS FOR NYCTANTHES ARBOR-TRISTIS STEM
|S. no.||Physicochemical parameter||Values (% w/w)|
|1||Moisture content||5.49 ± 0.02|
|2||Total ash||8.69 ± 0.17|
|3||Acid insoluble ash||0.21 ± 0.11|
|4||Water soluble ash||3.92 ± 0.05|
|5||Alcohol soluble extractive||15.93 ± 0.46|
|6||Water soluble extractives||18.31 ± 0.46|
TABLE 2: FLUORESCENCE ANALYSIS OF NYCTANTHES ARBOR-TRISTIS STEM
|Under UV light|
|Short wavelength (254 nm)||The long wavelength (365 nm)|
|Powder + Methanol||Brown||Light brown||Yellowish green|
|Powder + 70% ethanol||Brown||Light brown||Green|
|Powder + Pet. ether||Light brown||Light green||Green|
|Powder + 50% H2SO4||Brown||Greenish brown||Brownish|
|Powder + 50% HCl||Dark Brown||Green||Green black|
|Powder + 1N NaOH (aq.)||Light brown||Dark brown||Brownish black|
|Powder + 1N NaOH (alc.)||Light brown||Dark green||Greenish black|
|Powder + 50% HNO3||Brown||Light brown||Light green|
|Powder + 5% KOH||Brown||Purplish green||Dark purplish green|
|Powder + Ammonia||Brown||Green||Black|
|Powder + Picric acid||Yellowish brown||Green||Dark brown|
TABLE 3: PHYTOCHEMICAL SCREENING NYCTANTHES ARBOR-TRISTIS STEM
|S. no.||Class of constituents||PEE||CE||EAE||EE||AE|
PEE- Pet. ether extract, CE- Chloroform extract, EAE- Ethyl acetate extract, EE- Ethanol extract, AE- Aqueous extract, (-) Negative, (+) Positive
DISCUSSION: The wide use of herbal drugs in traditional medicines and herbal formulations, standardization that has been made to an important measure for ensuring and justifying the quality, purity, and authenticity of the crude drugs 31. Standardization purpose, morphological and microscopic analysis is one of the simplest and cheapest methods to start with establishing the correct identification of the source materials 32, 33. As there is no pharmacognostic work available on this medicinally potent plant, the present work was undertaken to lay down the standards which could be useful for establishing its authenticity. Organoleptic and microscopic studies are useful for identifying parameters for authentication of the drug 34, 35. Physicochemical studies act as a reliable method for detecting adulteration. Physicochemical constants like total ash, acid insoluble ash, and water soluble ash can serve as a valuable source of information which is usually helpful in the evaluation of purity and quality of a crude drug.
The earthly matter or inorganic composition and other impurities which are present along with the drug are determined by the ash values. Acid-insoluble ash usually indicates the contamination with silicon material like earth and sand. Water-soluble ash was used for the estimation of the amount of inorganic elements 36. The extractive values give an idea about the chemical constitution of the drug 33.
Fluorescence analysis is an alternative rapid useful method for the identification of authentic samples and recognizing adulterants. In this analysis, the crude drugs may be examined as such, in solution or as extracts and in powdered form 37. Fluorescence characteristics enable the identification and differentiation of plant materials from their adulterants when physical and chemical methods are scarce. Various chemical constituents present in the plant material exhibit fluorescence on absorbing light. Fluorescence is shown by some of the constituents even in the visible range in daylight. The ultraviolet light produces fluorescence in many natural products viz. alkaloids like berberine, which does not show fluoresce in the daylight. With the aid of different reagents, the nonfluorescent substances can easily be transformed into their fluorescent derivatives or decomposition products 33.
Phytochemical evaluation and chemo-profiling are useful for the quality assessment of plant materials. Phytochemical compounds in the plant are known to have various therapeutic importance. For instance saponins, terpenoids, flavonoids, tannins, steroids, and alkaloids have anti-inflammatory effects. Glycosides, flavonoids, tannins, and alkaloids have hypoglycemic activities 38, 39. Flavonoids possess the hepatoprotective, and antioxidant activities 40. The saponins have hypocholesterolemic and antidiabetic properties 41. In the animal studies, terpenoids tend to decrease blood sugar level. Steroids, as well as triterpenoids, exhibit the analgesic properties 42. The steroids and saponins are responsible for CNS activities 43.
CONCLUSION: The present study was focused on establishing pharmacognostic standards for the identification and authentication of the Nyctanthes arbor-tristis. Therefore, the outcomes of the above findings will serve as a promising source for laying down pharmacopoeial standards for future studies and research.
ACKNOWLEDGEMENT: We express our sincere thanks to Shri S. P. Singh Honorable Chairman, Sardar Bhagwan Singh Post Graduate Institute of Biomedical Sciences and Research, Balawala India for providing the facilities.
CONFLICT OF INTEREST: We declare that we have no conflict of interest.
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How to cite this article:
Joshi BC, Chauhan N, Sukanya and Uniyal S: Pharmacognostic and phytochemical evaluation of Nyctanthes arbor-tristis stem. Int J Pharmacognosy 2018; 5(6): 376-81. doi link: http://dx.doi.org/10.13040/IJPSR.0975-8232.IJP.5(6).376-81.
This Journal licensed under a Creative Commons Attribution-Non-commercial-Share Alike 3.0 Unported License.
B. C. Joshi *, N. Chauhan, Sukanya and S. Uniyal
Sardar Bhagwan Singh Post Graduate Institute of Biomedical Sciences and Research, Balawala, Dehradun, Uttarakhand, India.
16 February 2018
25 March 2018
30 March 2018
01 June 2018