Pharmacognostical study and establishment of quality parameters of aerial parts of Costus speciosus-a well known tropical folklore medicine

Pradeep Singh, Ratan Lal Khosa, Shruti Srivastava, Garima Mishra, Keshri Kishor Jha, Sourabh Srivastava, Sangeeta, Ramesh Kumar Verma, Mohd Adil Tahseen

1Department of Pharmacognosy, Teerthanker Mahaveer College of Pharmacy, Teerthanker Mahaveer University, Moradabad, Uttar Pradesh, India

2Department of Pharmacy, Bharat Institute of Technology, Partapur Bypass, Delhi Road, Meerut, Uttar Pradesh, India

Pharmacognostical study and establishment of quality parameters of aerial parts of Costus speciosus-a well known tropical folklore medicine

Pradeep Singh1*, Ratan Lal Khosa2, Shruti Srivastava1, Garima Mishra1, Keshri Kishor Jha1, Sourabh Srivastava1, Sangeeta1, Ramesh Kumar Verma1, Mohd Adil Tahseen1

1Department of Pharmacognosy, Teerthanker Mahaveer College of Pharmacy, Teerthanker Mahaveer University, Moradabad, Uttar Pradesh, India

2Department of Pharmacy, Bharat Institute of Technology, Partapur Bypass, Delhi Road, Meerut, Uttar Pradesh, India

PEER REVIEW

Peer reviewer

Antariksh Katara, National botanical research institute, Lucknow, India.

Tel: +91-8263904038

Email: katara.antariksh@gmail.com

Co-reviewer: Dr. Sachin Kumar, Moradabad, India.

Comments

The research carried out by authors on aerial parts of C. speciosus is genuine and valuable for the use of this plant as an ingredient in various formulations and treatment of several diseases because the authors have demonstrated all the quality control parameters, phytochemical analysis and microscopic characters which would be further authentic data in identification and authentication of this plant.

Details on Page 491

Objective:To evaluate the diagnostic pharmacognostical characters of Costus speciosus (aerial parts) along with their physico-chemical parameters and fluorosence analysis.

Costus speciosus, Quality control, Physico-chemical parameters, Microscopy, Fluorescence analysis

1. Introduction

Costus speciosusKoen., (C. speciosus) Costaceae, commonly known as keukand in Hindi; Kembuka in Sanskrit, is an erect, succulent, monocotyledonous perennial herb, up to 2.7 meters in height, arising from a horizontal rhizome, found in tropical region of India, and also cultivated for ornamental purposes[1]. It is ascribed to be bitter, astringent, acrid, cooling, aphrodisiac, purgative, anthelmintic, depurative and febrifuge in Ayurveda[2]. Rhizomes are used in the treatment of pneumonia, rheumatism, dropsy, urinary diseases and jaundice, whilethe leaves are used to treat mental disorders. Bruised leaves are applied in fever treatments, while a decoction of the stem is used in the treatment of fever and dysentery[1]. The species has been validated for certain pharmacological activities namely; antidiabetic and hypolipidemic activity and in the treatment of bronchial asthma[3-6], anticholinesterase activity[7], hepatoprotective activity[8], antioxidant activity[9,10], antibacterial activity, antifungal activity[11], antifertility activity[12] and spasmolytic activity. Phytoconstituents isolated from this species include: Diosgenin[13], furostanol saponins-costusosides I and J[12], β-sitosterol-β-D-glucoside, prosapogenins A and B of dioscin, dioscin, gracillin, dihydrophytylplastoquinone, α-tocopherolquinone and 5α-stigmast-9(11) en-3βol; methyl hexadecanoate, methyl octadecanoate and tetracosanyl octadecanoate[12], 24-hydroxytriacontan-26-one and 24-hydroxytriacontan-27-one[12]. Other components identified are 31-norcycloartanone, cyloartanol, cycloartenol and cycloalaudenol[14].

Irrespective of the recent phytochemistry and pharmacological research, till now no pharmacognostic work has been carried out on this species. The present investigation used different pharmacognostic and phytochemical parameters to supplement the identification and standardization information.

2. Materials and methods

2.1. Plant material

Fresh aerial parts (leaves) ofC. speciosusKoen. (20 plants), were collected from the Bahadurpur forests of Kolkata and authenticated byDr. D.C. Saini, Senior Scientist, Palaeobotany, Birbal Sahni Institute of Palaeobotany, Lucknow India. The voucher specimen no. BSIP 05 was deposited in Birbal Sahni Institute of Palaeobotany, Lucknow India. The leaves were dried under shade, mechanically reduced to moderate coarse powder and stored in amber colored air tight containers. Fresh leaves were used for the macroscopical and microscopical evaluation. Coarse form of drug was employed for determination of physicochemical parameters like moisture content, ash values, swelling index, foaming index, foreign organic matter, extractive values, qualitative and fluorescence analysis. The crude drug sample was also preserved in the department of Pharmacognosy, Teerthanker Mahaveer College of Pharmacy, Moradabad.

2.2. Macroscopic and microscopic analysis

The organoleptic characteristics such as shape, size, base and margin, venation of the leaves, taste and odour of the leaves ofC. speciosuswere determined. The macroscopy, microscopy and powder characteristics of aerial part of plant were studied according to the standard methods[15,16]. Leaf constantsviz.stomatal index and stomatal number were also studied according to the standard procedures[15]. Stomatal indices of both adaxial and abaxial epidermis in 1 mm2leaf surface was determined and recorded by using camera lucida and stage micrometer.

2.3. Physico-chemical analysis

Air dried leaves was used for the quantitative determination of ash values, extractive values, moisture content, swelling index, foaming index and foreign organic matter, via standard methods[15-19]. The total ash value for a crude drug is not always reliable, since there is a possibility of presence of non-physiological substances such as earthy matters. So, the parameters such as acid insoluble, water soluble and sulphated ash values were performed. Extractive values with petroleum ether, chloroform, ethyl acetate, ethanol and water were also determined. The fluorescence analysis is a tool for the determination of constituents in the plant that gives a definite idea of the chemical nature. Fluorescence analysis of crude powdered and various extracts was carried out by standard method[20].

2.4. Preliminary phytochemical screening

Preliminary phytochemical screening was performed by using standard procedures[15,16,21]. The extracts obtained from different solvents were subjected to identification tests for the detection of different phytoconstituents through organic and inorganic elements analysis, via the method of Khandelwal[15].

3. Results

3.1. Macroscopic characters

C. speciosusis an ornamental, perennial, erect, succulent herb, up to 2.7 m in height, arising from a horizontal rhizome. Leaves are alternate, spirally arranged, subsessile, elliptic to oblong or oblong-lancoelate, acuminate apex, thick, 15-35 cm×6-10 cm, glabrous above, silky pubescent beneath, with stem clasping sheaths up to 4 cm forming a complete tube around stem, flowers large, white, in thick, cone-like terminal spikes, with bright red bracts (Figure 1). Leaf lamina possessed 5 to 6 parallel veins on both sides of midrib No characteristic taste and odour was found (Table 1).

Figure 1. Aerial parts of C. speciosus.

Table 1 Organoleptic evaluation of leaves of C. speciosus.

3.2. Microscopic characters

3.2.1. Leaf

Transverse section of leaflet (Figure 2a, 2b, 2c, 2d) shows adaxial epidermis composed of single-layered polygonal cells, covered by a thick cuticle; containing tanniniferous substances and mucilage. Abaxial epidermis consists of bluntly angled cells with unicellular and multicellular uniseriate covering trichomes. Paracytic stomata are present on both adaxial and abaxial surface but more on abaxial surface. Mesophyll cells are distinguished between 2-3 layered loosely arranged spongy parenchyma cells and 2-layered palisade cells with shorter cells of lower layer than upper. Midrib is large and parenchymatous. The vascular bundles are amphicribal concentrici.e.phloem surrounds xylem; surrounded by double-layered sclerenchymatous cells.

Figure 2a. Transverse section of leaflet of C. speciosus viewed at 10× showing adaxial epidermis (UE) with thick cuticle followed by single layer of palisade cells (PC), abaxial epidermis (LE) with multicellular uniseriate trichomes (TR), Multilayered mesophyll cells (MC), vascular tissue with sclerenchymatous cells (SC).

Figure 2b. Transverse section of leaflet lamina of C. speciosus at 10× showing upper epidermis (UE) with cuticle, lower epidermis having multicellular uniseriate trichomes (TR), palisade cells, multilayered mesophyll cells (MS) and vascular bundles.

Figure 2c. Transverse section of leaflet lamina of C. speciosus at 40× showing vascular strands (VS) when treated with safranine.

Figure 2d. Section of lower surface of the leaflet (A) 10× view and (B) 40× view of trichomes (TR) and stomata (ST) clearly depicted the structure of an anomocytic stomata showing guard cells and subsidiary cells along with epidermal cells (EC).

3.2.2. Leaf constants

The results of leaf constants evinced that abaxial surface has stomatal number 0.15 and adaxial surface shows 0.06. Thus stomatal number are found to be more in abaxial surface than adaxial surface as shown in Table 2.

Table 2 Leaf constants of C. speciosus Koen.

3.2.3. Powder microscopy

Powder is dark green to brown, with no specific odour or taste. It is easily flowable with a fine to coarse texture. Diagnostic microscopic powder features include prismatic and clusters crystals of calcium oxalate; starch granules are mostly simple, but rarely compound; unicellular and uniseriate multicellular covering trichomes; parts of epidermis in surface view showing straight-walled polygonal epidermal cells, paracytic stomata and spiral xylem vessels (Figure 3).

Figure 3. Powdered characteristics of the aerial parts of C. speciosus showing A: Trichomes (TR) B: Xylem vessels (XV) C: Tetragonal Calcium oxalate crystals (CO) D: Epidermal cells (EC) E: Spiral xylem vessels (XV)

3.3. Physico-chemical analysis

The results of physico-chemical parameters such as total ash, acid insoluble ash, water soluble ash and sulphated ash are shown in Table 3. Sulphated ash value (12.5%) was lower than the total ash value (13.4%). The acid insoluble and water soluble ash values were 2.5% and 9%, respectively. Further, the results also showed that moisture content, swelling index and foaming index were found to be 1.43%, 0.7 cm and less than 100 respectively while foreign organic content was found to be nill (Table 4). The extractive values for various solvents such as petroleum ether, chloroform, ethyl acetate, ethanol and water were found to be 5%, 3%, 2%, 4% and 11% respectively (Table 5).

Table 3 Ash values of C. speciosus Koen.

Table 4 Moisture content, foreign organic matter, foaming index and swelling index of C. speciosus Koen.

Table 5 Extractive values of C. speciosus Koen.

The fluorescence characteristics of crude powdered drugs and various extracts were determined under UV radiation of long and short wavelengths and ordinary visible light. When the powdered drug and extracts were treated with different reagents and observed under UV and ordinary light, they emitted various colour radiations. The colour change for the crude powder and individual extract were distinctive and reproducible revealing the solvent properties to the phytoconstituents and data is presented in the Table 6.

3.4. Preliminary phytochemical screening

The phytochemical profile of the plant revealed the presence of alkaloids, carbohydrates, flavonoids, proteins, amino acids, phenols, tannins, glycosides and steroids as organic phytoconstituents (Table 7a). Qualitative analysis of various inorganic elements revealed the presence of iron, sulphates, phosphates and chlorides (Table 7b).

Table 6 Fluorescence studies of C. speciosus Koen.

Table 7a Phytochemical tests of C. speciosus Koen.

Table 7b Inorganic elements tests of C. speciosus Koen.

4. Discussion

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[22]. Standardization is an essential measurement for ensuring the quality control of the herbal drugs and also encompasses the entire field of study from birth of a plant to its clinical application.

C. speciosusKoen. (Costaceae) has many medicinal and therapeutic actions that have been scientifically validated and documented. The present investigation deals with all the physico-chemical and pharmacognostical perspectives of its leaves. The organoleptic evaluation depicts that leaves are alternate, spirally arranged, sub sessile, elliptic to oblong or oblong-lancoelate with no characteristic taste and odour whereas the microscopical analysis shows that leaves consists of adaxial and abaxial epidermis with paracytic stomata and covering trichomes. Mesophyll cells constitute palisade cells and spongy parenchyma along with amphicribal concentric vascular bundles surrounded by sclerenchyma. The diagnostic features of power microscopy include prismatic and clusters crystals of calcium oxalate; starch granules simple and spiral xylem vessels.

It was concluded from physico-chemical parameters that total ash value was found to be the highest (13.4%) while acid insoluble ash was lowest (2.5%). Cold extractive values revealed that the percentage yield of aqueous extract was highest 11.0% followed by petroleum ether (5.0%), ethanol (4.0%), chloroform (3.0%) and the lowest percentage yield was that of ethyl acetate (2.0%).

The moisture content was calculated through loss on drying method and was found to be 1.43%. The swelling and foaming index values were recorded to be 0.7 cm and less than 100 respectively. The phytochemical investigation indicate the presence of the organic phytoconstituents such as alkaloids, carbohydrates, flavonoids, proteins, amino acids, phenols, tannins, glycosides, steroids and inorganic constituents like iron, phosphates, sulphates and chlorides.

It was concluded from current investigation ofC. speciosusplant that the pharmacognostical data will provide the standards for its identification and authentication. The other parameters which are useful in the establishment of its quality control parameters are ash value, extractive values, moisture content, swelling and foaming index, leaf constants, fluorescence analysis and phytochemical parameters. The findings of the current research will help in the evaluation, identification and authentification of the plant.

Conflict of interest statement

We declare that we have no conflict of interest.

Acknowledgements

The authors are grateful to Prof. R. K. Mittal, Vice Chancellor, Teerthanker Mahaveer University, Moradabad for providing support and amenities to carry out this researchwork. The authors are also thankful to Dr. D. C. Saini, Senior Scientist, Palaeobotany, Birbal Sahni Institute of Palaeobotany, Lucknow India for plants identification and its authentication.

Comments

Background

In recent years, the use of herbal medicines has increased remarkably with the global trend of people returning to natural therapies because these are considered harmless and without side effects. Despite of all, quality evaluation of herbal preparation is a fundamental requirement of industry and other organization to ensure the purity and quality of herbal medicines.

Research frontiers

The aim of present research work is to investigate the various pharmacognostical and physico-chemical properties of aerial parts ofC. speciosus. Pharmacognostical studies includes macroscopy, microscopy, powder analysis and physico-chemical studies involved the assessment of ash values, moisture content, foaming index, swelling index, foreign organic content, extractive values and preliminary phytochemical screening.

Related reports

It is concluded from the literature survey thatC. speciosusis a traditional medicinal plant consisting of various therapeutic constituents and used in various diseases.

Innovations and breakthroughs

C. speciosuscommonly known as keukand in Hindi, is traditionally used as purgative, febrifuge, dysentery and as extensive acceptability as therapeutic agent for several diseases. Therefore, development of authentic analytical methods including pharmacognostical and physicochemical parameters is established to ensure the safety, efficacy and purity of this important plant.

Applications

The research carried out by authors can be applicable in identification of plant and engraining the therapeutic safety and efficacy.

Peer review

The research carried out by authors on aerial parts ofC. speciosusis genuine and valuable for the use of this plant as an ingredient in various formulations and treatment of several diseases because the authors have demonstrated all the quality control parameters, phytochemical analysis and microscopic characters which would be further authentic data in identification and authentication of this plant.

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10.12980/APJTB.4.2014C1103

*Corresponding author: Pradeep Singh, Department of Pharmacognosy, Teerthanker Mahaveer College of Pharmacy, Teerthanker Mahaveer University, Moradabad, Uttar Pradesh, India.

E-mail: pradeep_2682@yahoo.co.in

Tel: 09411072468

Article history:

Received 5 Mar 2014

Received in revised form 12 Mar, 2nd revised form 19 Mar, 3rd revised form 28 Mar 2014

Accepted 20 Apr 2014

Available online 28 Jun 2014

Method:The pharmacognostical characters were determined in terms of macroscopy, microscopy, powder microscopy, leaf constant, fluorescence analysis and preliminary phytochemical investigation.

Results:The findings of macroscopy revealed that leaves elliptic to oblong or oblong-lancoelate, thick, spirally arranged, with stem clasping sheaths up to 4 cm, flowers large, white, conelike terminal spikes, with bright red bracts. Transverse section of leaflet showed the presence of cuticularised epidermis with polygonal cells on adaxial surface and bluntly angled cells on abaxial surface of lamina, mesophyll cells differentiated in to single layered palisade cells on each surface and 2-3 layered spongy parenchyma, unicellular and uniseriate multicellular covering trichomes, paracytic stomata and vascular bundles surrounded by sclerenchymatous multicellular sheath. Preliminary phytochemical screening exhibited the presence of various phytochemical groups like alkaloids, glycosides, steroids, phenolic constituents. Further, the leaf constants, powder microscopy and fluorescence characteristics indicated outstanding results from this investigation

Conclusions:Various pharmacognostical and physico-chemical parameters have pivotal roles in identification, authentication and establishment of quality parameters of the species.