PHARMACOGNOSTICAL AND PRELIMINARY PHYTOCHEMICAL SCREENING OF THE ROOT TUBERS OF CURCULIGO ORCHIOIDES GAERTN.HTML Full Text
PHARMACOGNOSTICAL AND PRELIMINARY PHYTOCHEMICAL SCREENING OF THE ROOT TUBERS OF CURCULIGO ORCHIOIDES GAERTN.
Richa Tiwari * 1, 2 and Pankaj Gupta 1
School of Medical and Allied Sciences 1, K. R. Mangalam University, Sohna Road, Gurgaon - 122103, Haryana, India.
IIMT College of Pharmacy 2, Knowledge Park III, Plot No. 20A, Greater Noida - 201308, Uttar Pradesh, India.
ABSTRACT: India is well known as the emporium of medicinal plants harboring many plants of medicinal importance, of which Curculios orchioides is one that has been reported to possess medicinal importance for the treatment of various diseases traditionally as well as scientifically. An attempt was made herein to undertake detailed pharmacognostical and preliminary phytochemical investigation of the root tubers of Curculigo orchioides Gaertn. for the first time. The pharmacognostical study comprises of the taxonomical characters of the root tubers of Curculigo orchioides, histochemical tests and diagnostic characters. The phytochemical analysis consists of determination of physicochemical constants, fluorescence analysis, and preliminary organic analysis. The diagnostic characters of the plant showed that the leaves are radical with parallel venation while flowers are solitary, trimerous and ovary inferior. These parameters help in the standardization of the crude drug and in formulating Pharmacopoeial standards for the crude drug. Preliminary phytochemical screening of the crude drug showed the presence of carbohydrates and glycosides, phytosterols, fixed oils and fats, phenolic compounds and tannins, saponins, flavonoids, gums and mucilage.
Curculigo orchioides, Pharmacognostical, Phytochemical, Root tubers
INTRODUCTION: India is well known as the ‘Emporium of Medicinal Plants’. The use of plants to treat various diseases in India dates back to the times of Rigveda (3500 to 1800 B.C.). Later, the monumental Ayurvedic works like Charaksamhita 1 and Sushrutasamhita 2 followed by other Ayurveda and Siddha treatises have incorporated nearly 700 plant drugs entering into several medicinal preparations used in the management of health care. In fact these systems have been in practice even in remote areas of our country for centuries 3.
India has a well established indigenous system of medicine with rich materia medica which includes herbal, animal and mineral products. The use of herbs in India can be traced back to Vedic times; Rig veda (200-1000 B.C.) mentions the name of some medicinal herbs while the Atharva veda provides a more detailed account of herbs and the treatment of diseases. The most important and the authentic works on Indian medicinal plants is found in the classics of Ayurveda i.e., Charaksamhita 1, Susrutasamhita 2 and Astanga hridya 4 which are believed to have been written in the pre-Buddhist period, i.e., before 600 B.C.
These works incorporate 700 - 800 drugs of medicinal value used in several preparations for the treatment of various diseases. They also serve as the basis for the medicinal plant research in India and other countries.
Plants have contributed significantly to allopathic medicinal armory. Some of the drugs used today (e.g., aspirin, codeine, morphine, vinblastine, vincristine, pilocarpine, cocaine, atropine, emetine and ephedrine) have originated from medicinal plants 5. It has been estimated that approximately one fourth of the prescriptions dispensed from community pharmacies in United States contain one or more ingredients derived from plant origin 6. Around 80% of all contemporary pharmaceuticals are derived directly or indirectly from plant sources 7. According to World Health Organization as much as 80% of population in developing countries are dependent on plants for primary health care 8. Statistical data reveals that as many as 3226 out of 4752 communities in India are dependent on traditional medicine derived from plants 9.
A status report on Ethnobiology in India undertaken by Ministry of Environment and forests has indicated that tribal communities use over 7,500 species of plants for medicinal purposes 10. In India the total herbal products market, including sale of crude drugs, has been estimated at Rs. 2500 crore, of which 50% is contributed by Ayurvedic classical preparations and their modernized versions. This is a small percentage of the global trade estimated at $ 2000 crore. Thus, opportunities for developing modern phytomedicines based on leads from Ayurvedic drugs are indeed vast 11.
Curculigo orchioides known as Musali or Talamuli in Ayurveda is used in diseases traditionally as well as scientifically and root tuber is used for the treatment of diabetes, leukoderma, pain and aphrodisiac 12. The diagnostic characters of the plant showed that the leaves are radical with parallel venation while flowers are solitary, trimerous and ovary inferior. It is found in sub-tropical Himalayas from Kumaon estwards, Khasi Hills, Manipur, Bihar, West Bengal, Western Ghats, Konkan and Nilgiri Hills 13. An attempt was made herein to undertake detailed pharma-cognostical and preliminary phytochemical investigations of the root tubers of Curculigo orchioides for the first time 14, 15.
MATERIALS AND METHODS: The root tubers of Curculigo orchioides was collected from the Botanical garden of the University of Agricultural Sciences, GKVK Campus, Bangalore. The plant was authenticated at herbarium of the Regional Research Institute, Bangalore and authenticated by Dr. Yoganarasimhan and a voucher specimen HS1704990020 has been retained at the School of Medical and Allied Sciences, K. R. Mangalam University, Gurgaon. The root tubers were washed with water and dried in the laboratory at room temperature (30-40 °C). The roots were ground to pass a sieve of 1 mm and further subjected to analytical parameters like ash value and extractive value.
Pharmacognostical Studies: A small quantity of the fresh root tuber was collected and preserved in tubes containing 70% alcohol for pharma-cognostical work. The macroscopical observations were carried out following reported procedures 16. The microscopical investigations, histochemical tests were carried out following the given procedures 17. For microscopical studies free hand sections were taken using a sharp razor. The sections were cleared by warming with a few drops of chloral hydrate, stained with phloroglucinol: HCl (1:1), treated with iodine solution and safranin. The sections were mounted temporarily in glycerine for microscopical observations. The measurement of tissues was recorded using stage and ocular micrometers, (Erma-Japan).
Physicochemical Constants and Preliminary Phytochemical Screening: Physical constants, phytochemical tests, fluorescence studies were carried using reported procedures 18, 19, 20, 21, 22, 23. The successive extracts obtained were subjected to qualitative tests for the identification of various plant constituents. For fluorescence studies, drug was sieved through 60 mesh and observations were made following the reported procedures 21, 22, 23.
Microscopical Characters: A transverse section of root tuber is circular in outline and shows the following characters Fig. 1. Outer peripheral cortical portion having a starchy lustre and a central stellar region (3 mm in diameter) wherein most of the vascular bundles are located. The outermost tissue is the cork consisting of 5- 8 rows of cells with light brown coloured cell walls. In young tubers the cells of this tissue are cubical to rectangular in shape but in older tubers the cells are tangentially elongated.
FIG. 1: TRANSVERSE SECTION OF ROOT TUBERS OF CURCULIGO ORCHIOIDES; a- CORK, b- CORTEX, c-LYSIGENOUS CAVITY, d- STELE
Next to the cork is the cortex containing 30 to 40 rows of large collenchymatous cells with many intercellular spaces. These cells are abundantly packed with starch grains. They are either single or in groups of 2 to 3. Some of the cortical cells are larger, rounded or oblong and contain bundles of raphides of calcium oxalate crystals. The raphides are acicular in shape. Within the cortex are found rounded to tangentially elongated lysigenous cavities. Small globules of mucilage are found within these cavities In mature tubers the cortical region is differentiated into a narrow outer zone (0.5-1 mm in thickness) wherein starch grains are absent and an inner wider starchy zone. The cells of the outer zone resemble those of the inner in shape but their walls have nearly double the thickness of the latter and are slightly lignified. A few scattered vascular bundles are also observed in cortex sometimes. Endodermis and pericycle are indistinct but the stele is clearly demarcated from the cortex in having ground tissue made up of parenchymatous cells. The stele is crescent shaped and consists of numerous closed, collateral vascular bundles. The outer bundles are arranged in a ring while the inner are found scattered in ground tissue. The vascular bundles vary in size and shape and range from collateral to the amphivasal type. The xylem elements partly or completely surround the phloem. The cells of phloem tissue are very small and are polygonal; sieve tubes and companion cells are indistinct. A parenchymatous pith is present in the centre of the stele. The measurement of various cells are provided in Table 1.
TABLE 1: MEASUREMENT OF CELLS OF DIFFERENT TISSUE (IN μm)
|Cells (T. S.)||Minimum||Average||Maximum|
Histochemical Tests: The sections were treated with different reagents and results are provided in Table 2.
TABLE 2: TREATMENT OF SECTIONS WITH DIFFERENT REAGENTS
|1. Section||Iodine solution||Starch||Blue colour||+|
|2. Section||Ferric chloride||Tannin||Black colour||+|
|3. Section||Sudan 3 solution||Oil globule||-||-|
|4. Section||Conc. HCl||Crystals||Effervescence||+|
|5. Section||Pinch of Phloroglucinol + dil. HCl||Lignin||Magenta colour||+|
+ = present; - = absent
Phytochemical Analysis: The organic and inorganic constituents present in a drug or plant play a significant role in the identification of crude drug. The physical constants like ash and extractive values help in establishing the Pharmacopoeial standards of the drug.
The fluorescence analysis help to identify the drug in powder form and the organic analysis help to understand the organic constituents of the drug.
Physical Constants: The results of moisture content, ash and extractive values are presented in Tables 3, 4, 5 respectively.
TABLE 3: MOISTURE CONTENT OF THE ROOT TUBER OF CURCULIGO ORCHIOIDES GAERTN.
|Fresh weight (g)||Dry
|Loss on drying (g)||Moisture Content (%)|
TABLE 4: ASH VALUES OF THE ROOT TUBER OF CURCULIGO ORCHIOIDES GAERTN.
|Acid insoluble ash %||Water soluble ash %|
TABLE 5: EXTRACTIVE VALUES OF THE ROOT TUBER OF C. ORCHIOIDES GAERTN.
|S. no.||Extractives||Extractive values (% w/w)|
Fluorescence Analysis: Fluorescence provided by a drug is one of the several methods used for analyzing crude drugs. Fluorescence is a type of luminescence in which the molecule emits visible radiation passing from a higher to lower electronic state. The results are presented in Table 6.
TABLE 6: FLUORESCENCE ANALYSIS OF THE ROOT TUBER OF CURCULIGO ORCHIOIDES GAERTN.
|Short wave (254 nm)||Long wave (365 nm)|
|Powder as such||Light Brown||Light brown||Light brown|
|In methanol||Dark brown||Light green||Dark brown|
|In methanolic NaOH||Green||Dark green||Green|
|In ethanol||Dark brown||Light green||Light green|
|In ethanolic NaOH||Green||Dark Green||Dark green|
|In dil. HCl||Light brown||Dark green||Dark green|
Successive Solvent Extraction: The results of Successive solvent extraction were presented in Table 7.
Preliminary Organic Analysis: The results of Preliminary organic analysis were presented in Table 8.
TABLE 7: SUCCESSIVE SOLVENT EXTRACTIVE VALUES AND NATURE OF EXTRACTS OF CURCULIGO ORCHIOIDES GAERTN.
|S. no.||Solvent||Colour||Consistency||Extractive value(%w/w)|
|1||Petroleum ether (60-80 °C)||Pale yellow||Lumpy mass||2.161|
TABLE 8: PRELIMINARY PHYTOCHEMICAL ANALYSIS OF THE ROOT TUBER OF CURCULIGO ORCHIOIDES GAERTN.
|Tests||Pet. ether extract||Benzene extract||Chloroform
|Carbohydrates & Glycosides||-||-||-||+||+||+|
|Fixed oil and Fats||+||+||-||-||-||-|
|Phenolic compound & Tannins||-||-||_||+||+||-|
|Proteins & Amino acids||-||-||-||-||-||-|
|Gums and Mucilage||-||-||-||-||-||+|
+ = present; - = absent
DISCUSSION: The root tuber which is the part used can be identified by the following macro and microscopical characters - tuberous root with starchy cortex; presence of bundles of raphides made of acicular crystals; presence of lysigenous cavity in the ground tissue; crescent shaped vascular bundles; presence of pith.
Histochemical tests show the presence of starch, tannin, crystals and lignin. The physicochemical constants of the root tuber of Curculigo orchioides were determined. It was found that water soluble extractive (13.145% w/w) was more than alcohol soluble extractive value (9.506% w/w). These help in formulating pharmacopoeial standards for the drug.
Fluorescence analysis helps in distinguishing the drug in powder form. In case of successive solvent extraction the quantity of extract was found to be maximum in case of water extract and minimum in case of chloroform extract. The extract obtained by successive solvent extraction were subjected to preliminary phytochemical analysis, which revealed the presence of carbohydrates and glycosides, phytosterols, fixed oils and fats, phenolic compounds and tannins, saponins, flavonoids gums and mucilage. The preliminary organic analysis of drug helps to undertake further studies on the isolation and identification of specific chemical constituents.
CONCLUSION: Curculigo orchioides Gaertn. (Family Hypoxidaceae) commonly known as Musali or Talamuli in Ayurveda is an important drug of Ayurveda and Siddha system of medicine. The root tuber is used in disease like diabetes, pain, leukoderma, skin troubles, demulcent and useful in diarrhoea. In the present study pharmacognostical, phytochemical investigations on the root tuber of C. orchioides were carried out. It is concluded that scientific parameters based on taxonomical, pharmacognostical and phytochemical studies are essential to identify the plant.
These parameters not only help in the standardization of these drugs but also help in formulating pharmacopoeial standards for drugs. The exomorphic characters have been found to be useful tools to identify the species taxonomically. The phytochemical analysis helps to identify the drug in powder form on the basis of physical constants and fluorescence analysis.
CONFLICT OF INTEREST: Nil
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How to cite this article:
Tiwari R and Gupta P: Pharmacognostical and preliminary phytochemical screening of the root tubers of Curculigo orchioides Gaertn. Int J Pharmacognosy 2018; 5(12): 788-92. doi link: http://dx.doi.org/10.13040/IJPSR.0975-8232.IJP.5(12).788-92.
This Journal licensed under a Creative Commons Attribution-Non-commercial-Share Alike 3.0 Unported License.
R. Tiwari * and P. Gupta
School of Medical and Allied Sciences, K. R. Mangalam University, Sohna Road, Gurgaon, Haryana, India.
12 October 2018
22 November 2018
27 November 2018
01 December 2018