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ORIGINAL ARTICLE
Year : 2016  |  Volume : 4  |  Issue : 1  |  Page : 19-25

Establishment of quality parameters and pharmacognostical study of Cyperus rotundus Linn. (Cyperaceae): A well-known traditional medicinal plant


1 Department of Pharmacognosy, Teerthanker Mahaveer College of Pharmacy, Teerthanker Mahaveer University, Moradabad, Uttar Pradesh, India
2 Department of Pharmaceutical Sciences, Bharat Institute of Technology, Meerut, Uttar Pradesh, India
3 Department of Pharmaceutical Chemistry, Teerthanker Mahaveer College of Pharmacy, Teerthanker Mahaveer University, Moradabad, Uttar Pradesh, India

Date of Web Publication2-Jul-2018

Correspondence Address:
Dr. Pradeep Singh
Department of Pharmacognosy, Teerthanker Mahaveer College of Pharmacy, Teerthanker Mahaveer University, Moradabad, Uttar Pradesh
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/njecp.njecp_20_16

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  Abstract 


Objective: Cyperus rotundus (L.) (family-Cyperaceae) occupies a prime role in traditional system of medicine. Since the extensive literature survey did not provide proper and accurate information regarding its study on standardization; therefore, the current study is carried out to establish various quality control parameters of C. rotundus rhizomes. Materials and Methods: The investigation included the determination of various standardization parameters such as macroscopic and microscopic studies, physicochemical parameters as well as phytochemical analysis of crude drug. Results: The microscopy study revealed that rhizome of the plant shows typical monocotyledonous characters with epidermal layer, sclerenchymatous layers, cortex, collateral, closed, vascular bundles, and endodermis. Physicochemical constants such as moisture content, ash values, fluorescence analysis, and extractive values were established. Preliminary phytochemical analysis confirmed the presence of alkaloids, flavonoids, glycosides, saponins, tannins, etc. Conclusion: Various pharmacognostical and physicochemical parameters have pivotal roles in identification, authentication, and establishment of quality parameters of the species.

Keywords: Cyperaceae, Cyperus rotundus, nagarmotha, physicochemical properties, standardization


How to cite this article:
Singh P, Khosa RL, Garima M, Jha KK. Establishment of quality parameters and pharmacognostical study of Cyperus rotundus Linn. (Cyperaceae): A well-known traditional medicinal plant. Niger J Exp Clin Biosci 2016;4:19-25

How to cite this URL:
Singh P, Khosa RL, Garima M, Jha KK. Establishment of quality parameters and pharmacognostical study of Cyperus rotundus Linn. (Cyperaceae): A well-known traditional medicinal plant. Niger J Exp Clin Biosci [serial online] 2016 [cited 2018 Sep 19];4:19-25. Available from: http://www.njecbonline.org/text.asp?2016/4/1/19/235805




  Introduction Top


The genus Cyperus includes common weeds found in upland and paddy fields in temperate to tropical regions. In Asian countries, the rhizomes of Cyperus rotundus (L.) (family-Cyperaceae), which are used as traditional folk medicines for the treatment of stomach and bowel disorders, and inflammatory diseases, have been widely, investigated.[1]

C. rotundus (L.) (family-Cyperaceae), commonly known as Nutgrass in English and Motha in Hindi is native to Africa, southern and central Europe, and Southern Asia. It is a perennial plant that may reach a height of up to 40 cm. The names “nut grass” and “nut sedge” (shared with the related species Cyperus esculentus) are derived from its tubers that somewhat resemble nuts, although, botanically they have nothing to do with nuts. It prefers a moist sandy loam [2],[3] and a sunny position.[4],[5] This plant is sometimes cultivated for its edible tubers in tropical regions.[6] However, it is a serious weed of agricultural land in the tropics, where it spreads rapidly at the roots [7] and is considered to be one of the world's most damaging weeds.[4],[8],[9]

This plant is known to possess carminative, astringent, diuretic, emmenagogue [4],[10],[11],[12] anti-inflammatory, antirheumatic, hepatoprotective, antipyretic, analgesic, hypotensive, and nervine tonic.[10] It is also good for intestinal problem, indigestion, sprue, dysentery, vomiting and fever, also as a hypocholesterolemic drug and in obesity,[10] diarrhea,[11] aromatic, contraceptive, lithontripic, sedative,[4] diaphoretic, stimulant, stomachic, tonic, vermifuge [4],[11],[12] and demulcent.[11],[12] In addition, it has been reported for antimalarial,[13] antibacterial, antispasmodic, antitussive, and anthelmintic activities.[11]

The present investigation reports different pharmacognostic and phytochemical parameters to supplement the identification and standardization information.


  Materials and Methods Top


Plant material

Fresh rhizomes of the plant were collected from the local area of Moradabad and authenticated by Dr. DC Saini, Senior Scientist, Palaeobotany, Birbal Sahni Institute of Palaeobotany, Lucknow, India. The voucher specimen (BSIP 03) was deposited in Birbal Sahni Institute of Palaeobotany, Lucknow, India. The crude drug sample was also preserved in the Department of Pharmacognosy, Teerthanker Mahaveer College of Pharmacy, Moradabad. The rhizomes were dried under shade, mechanically reduced to moderate coarse powder, and stored in amber-colored airtight containers. Fresh rhizomes were used for macroscopical and microscopical evaluation, while coarse form of drug was employed for the determination of physicochemical parameters such as moisture content, ash values, swelling index, foaming index, foreign organic matter, extractive values, qualitative, and fluorescence analysis.

Macroscopic and microscopic analysis

The organoleptic characteristics such as color, odor, taste, shape, and size of C. rotundus rhizomes were determined. The macroscopy, microscopy, and powder characteristics of rhizomes of the plant were studied according to the standard methods.[14],[15],[16]

Determination of physicochemical parameters

Foreign organic matter

The drug sample to be examined was weighed and spread on a white tile uniformly without overlapping. The foreign matter was separated manually and examined in daylight with unaided eye. The suspected particles were transferred into a  Petri dish More Details. After complete separation, weight of the foreign matter was taken and the percentage (%) (w/w) was determined.[14],[17]

Moisture content

A sample of 10 g of drug (without preliminary drying) after accurately weighing was placed in a tared evaporating dish and dried at 105°C. The drying and weighing were continued at 1 h interval until difference between two successive weighing corresponded to not <0.25%. A constant weight was supposed to have reached when two consecutive weighing after drying for 30 min and cooling for 30 min in a desiccator showed not <0.01 g difference.[14],[15],[16],[17],[18],[19],[20],[21]

Swelling index

Specified quantity of plant material was introduced into a 25 ml glass-stoppered measuring cylinder. A volume of 25 ml of water was added, and the mixture was shaken thoroughly every 10 min for 1 h. The crude drug was allowed to stand for 3 h at room temperature. The volume occupied by the plant material was measured in milliliter including any sticky mucilage. The mean value of the individual determination was finally calculated relative to 1 g of plant material.[14],[21]

Foaming index

A total of 1 g of the plant material was reduced to a coarse powder, weighed accurately, and transferred to a 500 ml conical flask containing 100 ml of boiling water. It was maintained at moderate boiling for 30 min, cooled, and filtered into a 100 ml volumetric flask. Sufficient water was added through the filter to dilute the filtrate to make up the volume. The decoction was poured in to 10 stoppered test tubes in successive portions, i.e., 1, 2, and 3 ml and the volume of liquid in each test tube was adjusted with water to 10 ml. The tubes were stoppered and shaken in a lengthwise motion for 15 s at the rate of two shakes per se cond and allowed to stand for 15 min. The height of the foam was measured, and the result was calculated.[21]

Ash values

Total ash

About 2.0 g of crude drug was accurately weighed and incinerated in a silica crucible at a temperature not exceeding 450°C until free from carbon. The resulting ash was then cooled and weighed. The procedure was repeated to obtain constant weight. The percentage of total ash with reference to the air-dried drug was finally calculated.[14],[15],[16],[17],[18],[19],[20]

Acid insoluble ash

To the crucible containing total ash, 25 ml of dilute hydrochloric acid was added. The insoluble matter was collected on an ashless filter paper. It was then washed with hot water until it became neutral and ignited to a constant weight. The residue was allowed to cool in a suitable desiccator for 30 min and it was immediately weighed. The procedure was repeated to obtain constant weight. The percentage of acid-insoluble ash with reference to the air-dried drug was finally calculated.[14],[15],[16],[17],[18],[19],[20]

Water soluble ash

To the crucible containing total ash, 25 ml of water was added and boiled for 5 min. The insoluble matter was collected on an ashless filter paper. It was then washed with hot water and ignited for 15 min at a temperature not exceeding 450°C. The procedure was repeated to obtain constant weight. The difference in the weight of ash and weight of insoluble matter was calculated. The percentage of water-soluble ash with reference to the air-dried drug was finally determined.[14],[15],[16],[17],[18],[19],[20]

Sulfated ash

Silica crucible was heated to redness for 10 min and it was allowed to cool in a desiccator and weighed. 1–2 g of crude drug was accurately weighed and was put into the crucible. It was ignited gently at first so that it could thoroughly charred, then cooled and moistened with 1 ml of sulfuric acid. The sample was heated gently at first until white fumes ceased off and later on ignited at a temperature of 800°C ± 25°C until all black disappeared. The crucible was then allowed to cool and again few drops of sulfuric acid was added and heated again. The steps were repeated until two successive weighing did have the difference of weight not <0.5 mg.[14],[15],[16],[17],[18],[19],[20]

Cold extractive values

Extractive values for the plant drug sample were determined using different solvents, i.e., petroleum ether, chloroform, ethyl acetate, ethanol, and water.[14],[15],[16],[17],[18],[19],[20]

Alcohol soluble extractive

A total of 5 g of air-dried drug was first coarsely powdered and macerated with 100 ml of alcohol of specified strength in a closed flask for 24 h. The flask was shaken frequently during 6 h and allowed to stand for 18 h. The extract was filtered rapidly and evaporated up to 25 ml rapidly to dryness in a china dish. Further, it was dried to obtain a constant weight. The percentage yield of alcohol-soluble extractive with reference to air-dried drug was finally calculated. Similar procedure was followed with petroleum ether, chloroform, ethyl acetate, and water to find out their respective extractive values.

Hot extractive values

The rhizome of C. rotundus was made into coarse powder which was further extracted with soxhlet apparatus using petroleum ether (60°C–80°C) and ethanol. The extracts were concentrated under reduced pressure and evaporated to dryness on water bath. The extracts were stored in desiccator for phytochemical analysis.

Fluorescence analysis

A small quantity of dried and finely powdered crude drug was placed on a grease free clean microscopic slide and the same was treated with 1–2 drops of freshly prepared reagent solutions separately, i.e., 1 N sodium hydroxide in methanol, 1 N sodium hydroxide in water, 50% sulfuric acid, 50% nitric acid, and 1N HCl. At each effort, the added reagent was mixed by gentle tilting the slide and waited for 1–2 min. Then, each slide was placed inside the ultraviolet (UV) chamber and viewed in natural and UV lights. All extracts obtained were subjected to fluorescence analysis. The colors observed by the application of different reagents were recorded.[22],[23]

Preliminary phytochemical investigation

Determination of organic phytoconstituents

Petroleum ether and ethanolic extracts were subjected to preliminary phytochemical analysis to identify the various organic phytoconstituents such as alkaloids, glycosides, saponins, flavonoids, tannins, and steroids.[14],[17],[24]

Determination of Inorganic Constituents

Ash of drug material was treated with 50%v/v HCl, kept for 1 h and filtered. The filtrate was used for qualitative determination of various inorganic constituents such as calcium, magnesium, potassium, and iron.[14],[17]


  Results Top


Macroscopic evaluation

Macroscopic characters of rhizomes of C. rotundus are as shown in [Table 1].
Table 1: Organoleptic evaluation of Cyperus rotundus rhizomes

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Microscopic evaluation

Transverse section of rhizome

Transverse section of rhizome as shown in [Figure 1]a, [Figure 1]b, [Figure 1]c, [Figure 1]d, [Figure 1]e, [Figure 1]f, [Figure 1]g, [Figure 1]h, [Figure 1]i revealed the presence of single layered epidermis followed by 2–3 layered suberised tubular sclerenchymatous cells; both the layers are filled with dark brown content. Ground tissue (cortex) consists of circular to oval, thin-walled parenchymatous cells with small intercellular spaces. Numerous collateral, closed, vascular bundles (VBs) and clusters type calcium oxalate crystals are scattered in this region. A distinct and definite endodermis surrounding the stele is present. Smaller VBs are occurs near to endodermis while larger VBs are found towards the center, scattered throughout the ground tissue. Simple round-to-oval starch grains and few pigmented cells filled with reddish brown content are present throughout ground tissue and steler region.
Figure 1: Cyperus rotundus (L.) plant and rhizomes. (a) Transverse Section (TS) of rhizome of Cyperus rotundus at ×10 showed single-layered epidermal cells cortex, calcium oxalate cluster and collateral, closed vascular bundle. (b) TS of rhizome of Cyperus rotundus at ×10 showed cortex region, calcium oxalate cluster and collateral, closed vascular bundles. (c) TS of rhizome of Cyperus rotundus at ×10 showed single-layered endodermis, sclerenchyma, cortex, calcium oxalate cluster and scattered collateral, closed vascular bundle. (d) TS of rhizome of Cyperus rotundus at ×40 showed single-layered epidermal cells, sclerenchyma, cortex, and starch grains. (e) TS of rhizome of Cyperus rotundus at ×40 showed single-layered epidermal cells, sclerenchyma, cortex, and starch grains. (f) TS of rhizome of Cyperus rotundus at ×40 showed cortex, collateral, closed vascular bundle, and starch grains. (g) TS of rhizome of Cyperus rotundus at ×40 showed cortex and collateral, closed vascular bundle. (h) TS of rhizome of Cyperus rotundus at ×40 showed pigmented cell and collateral, closed vascular bundle. (i) TS of rhizome of Cyperus rotundus at ×40 showed calcium oxalate cluster and starch grains

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Powder microscopy of rhizome

Diagnostic characters of powder microscopy of rhizome shows the presence of clusters calcium oxalate crystals; circular or oval starch grains; parenchymatous cells filled with volatile oil, pigmented cells, and scalariform xylem vessels [Figure 2]a, [Figure 2]b, [Figure 2]c, [Figure 2]d, [Figure 2]e.
Figure 2: (a) Powdered characteristics of rhizome of Cyperus rotundus at ×40 showed calcium oxalate cluster, and starch grains. (b) Powdered characteristics of the rhizome of Cyperus rotundus at ×40 showed oil cells (parenchymatous cells filled with volatile oil) and pigmented cells. (c) Powdered characteristics of rhizome of Cyperus rotundus at ×40 showed scalariform xylem vessels. (d) Powdered characteristics of the rhizome of Cyperus rotundus at ×40 showed scalariform xylem vessels. (e) Powdered characteristics of the rhizome of Cyperus rotundus at ×40 showed epidermal cells and starch grains

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Physico-chemical evaluation

The results of physicochemical parameters are summarized in [Table 2].
Table 2: Physicochemical values of Cyperus rotundus rhizomes

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Foreign organic content

The foreign organic content was found to be 1%w/w.

Moisture content

Moisture content of rhizomes of C. rotundus was found to be 10.56%w/w.

Swelling and foaming index

The swelling index was observed as 0.4 cm while foaming index was <100.

Ash values

Ash value gives idea about inorganic content as well as extraneous matter (e.g., soil and sand) cling to the surface of plant. Total ash, acid insoluble ash, water soluble ash, and sulfated ash of the rhizomes of C. rotundus were recorded as 10, 3.5, 6, and 9.1%w/w, respectively.

Extractive values

Cold extractive values for solvents, i.e., petroleum ether, chloroform, ethyl acetate, ethanol, and water were noted as 2.5, 2, 2, 8, and 15.5%w/w, respectively. Extractive values using soxhlet apparatus for solvents such as petroleum ether and ethanol in successive order were recorded as 4.4% and 10.5%w/w, respectively.

Fluorescence analysis

The results of fluorescence analysis are given in [Table 3].
Table 3: Fluorescence analysis of Cyperus rotundus rhizomes

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Phytochemical analysis

Preliminary phytochemical screening of petroleum ether and ethanolic extract showed the presence of alkaloids, flavonoids, tannins, glycosides, saponins, and steroids as organic phytoconstituents [Table 4]a. Qualitative analysis of various inorganic elements revealed the presence of calcium, iron, sulfate, phosphates, and chlorides [Table 4]b.


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  Discussion Top


The quality control of crude drugs and herbal formulation is of paramount importance in justifying their acceptability in modern system of medicine, but one of the major problems faced by the herbal drug industry is nonavailability of rigid quality control profile for herbal material and their formulations. Standardization is an essential measurement for ensuring the quality control of the herbal drugs. It encompasses the entire field of study from birth of a plant to its clinical application.[16]

In the present investigation, various pharmacognostical and physicochemical parameters such as ash values, extractive values, and moisture content of rhizome of C. rotundus were established, which are important tools for determination of the identity, quality, and purity of the drug. Transverse section of rhizome confirmed the presence of epidermis, sclerenchyma, cortex, collateral, closed, VBs, and endodermis which shows histological characters of typical monocotyledonous rhizome. Moisture in conjunction with a suitable temperature lead to the activation of enzymes and gives suitable condition to the proliferation of living organism which is a major factor responsible for the deterioration of the drugs and formulations.[20] Swelling index is the property of medicinal plant containing gums, mucilage, pectin, or hemicelluloses which may be responsible for specific therapeutic or pharmaceutical utility, while foaming index determines the saponin content of the drug.[17] Total ash indicates the presence of inorganic salts like phosphates, carbonates and silicates of sodium, potassium, magnesium, calcium, etc. Acid insoluble ash is the residue obtained after boiling the ash with hydrochloric acid and igniting the insoluble portion. It gives measure of sand and other silicious matter. Water-soluble extractive value shows the presence of water soluble inorganic salts. Sulfated ash value gives more consistent data by treatment of ash with sulfuric acid, which converts all the oxides and carbonates to sulfates.[17],[20] Phytochemical standardization encompasses all possible information regarding the chemical constituents present in an herbal drug. Preliminary phytochemical analysis showed the presence of flavonoids and steroids in petroleum ether extract and alkaloids, glycosides, saponins, flavonoids, and tannins in ethanolic extract of rhizome of C. rotundus.


  Conclusion Top


In the present investigation, different quality control parameters, including pharmacognostical, physicochemical, and phytochemical, were established, which might be useful in identifying the authentic drug and setting the pharmacopoeial standards for further studies.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
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    Figures

  [Figure 1], [Figure 2]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4]



 

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