ORIGINAL ARTICLE PHCOG J. 32 Pharmacognosy Journal | September 2011 | Vol 3 | Issue 25 *Address for correspondence: Senior Research Fellow, Central Council For Research In Ayurveda & Siddha, Dept. of AYUSH, Ministry of Health & Family Welfare, Janakpuri, New Delhi-58, Ph: +919968532138 (M) E-mail: [email protected]DOI: 10.5530/pj.2011.25.6 where botanical identity of plant source has not been established and these are subjected to adulterations/ substitutions. So the aim of the present study was focussed on the pharmacognosy, and phytochemical properties, which of the drug from various substitutes and adulterants and its comparison with market samples collected in the name of Kakoli and Ksheerkakoli to standardize and maintain the ‘quality control’ the drug. MATERIAL AND METHODS Roscea procera Wall. and Lilium polyphyllum D. Don were collected from medicinal plants garden of Regional Research Institute of Himalayan Flora, Tarikhet and Indian Medicines Pharmaceutical Corporation Limited, Mohan (Almora). Tubers were washed, cut into pieces and preserved in Formalo-acetyl-alcohol (FAA) and labelled RP4 and LP4 for pharmacognostical study. Microtome section were taken, stained and mounted following the usual plant microtechniques [9,10] and representative diagram sketched through camera lucida and some were shade dried and coarse (20 to 30 #) powdered for qualitative tests and physico-chemical study as per IP/API / WHO guidelines. The physicochemical parameters like total ash, acid insoluble ash, water and alcohol soluble extractives Pharmacognostical & Phytochemical Studies of Roscea procera (Kakoli) and Lilium Polyphyllum (Ksheerkakoli) in comparison with market samples Rath Chinmay*, Suman Kumari # , Dhar Bishnupriya # , Mohanty RC ## , Dixit Renu # , Padhi MM # , Babu Ramesh # # Central Council for Research in Ayurveda & Siddha, Dept. of AYUSH, Ministry of Health & Family Welfare, Janakpuri, New Delhi-58. ## Department of Botany, Utkal University, Vani Bihar, Bhubaneswar, Orissa ABSTRACT Kakoli (Roscea procera Wall.) of Family Zingiberaceae and Ksheerkakoli (Lilium polyphyllum D. Don) of family Liliaceae, are mentioned as drugs of Astavarga, but the botanical identity of both the species are controversial and different drugs are sold/ used in the name of Kakoli and Ksheerkakoli. This has created frightening problem with made to lay down pharmacopoeial characters and to identify botanical identity of market samples by comparision with genuine drugs. Key words: Macroscopy, Microscopy, Thin Layer Chromatography, physico chemical evaluation. INTRODUCTION The identity of Astavarga drugs suffer a lot of confusion in Ayurvedic literature. According to various Ayurvedic Nighantu books composed or commented by different Vaidyas, it constitutes a group of eight drugs, which form an important constituent of a number of Ayurvedic preparations. These are known in Sanskrit as Jivaka, Rishibhak, Mahameda, Meda, Kakoli, Ksheerkakoli, Riddhi and Vriddhi. [1-3] Bhav Mishra in his Bhav Prakash Nighantu has further mentioned that none of Astavarga drugs are true. [4] Kakoli and Ksheerkakoli have been described under “Brhneeya” (the drugs which promote the formation of mansadhatu [5] Caraka Samhita, [6] Sushruta Samhita [7] and Astanghridaya. [8] Till date none has tried to work out on their pharmacognosy and chemistry evaluation therefore they need further investigation. There is however a number of crude drugs RETRACTED
*Address for correspondence:Senior Research Fellow, Central Council For Research In Ayurveda & Siddha, Dept. of AYUSH, Ministry of Health & Family Welfare, Janakpuri, New Delhi-58,Ph: +919968532138 (M)E-mail: [email protected]
DOI: 10.5530/pj.2011.25.6
where botanical identity of plant source has not been established and these are subjected to adulterations/substitutions. So the aim of the present study was focussed on the pharmacognosy, and phytochemical properties, which
of the drug from various substitutes and adulterants and its comparison with market samples collected in the name of Kakoli and Ksheerkakoli to standardize and maintain the ‘quality control’ the drug.
MATERIAL AND METHODS
Roscea procera Wall. and Lilium polyphyllum D. Don were collected from medicinal plants garden of Regional Research Institute of Himalayan Flora, Tarikhet and Indian Medicines Pharmaceutical Corporation Limited, Mohan (Almora). Tubers were washed, cut into pieces and preserved in Formalo-acetyl-alcohol (FAA) and labelled RP4 and LP4 for pharmacognostical study. Microtome section were taken, stained and mounted following the usual plant microtechniques[9,10] and representative diagram sketched through camera lucida and some were shade dried and coarse (20 to 30 #) powdered for qualitative tests and physico-chemical study as per IP/API / WHO guidelines. The physicochemical parameters like total ash, acid insoluble ash, water and alcohol soluble extractives
Pharmacognostical & Phytochemical Studies of Roscea procera (Kakoli) and Lilium Polyphyllum (Ksheerkakoli)
#Central Council for Research in Ayurveda & Siddha, Dept. of AYUSH, Ministry of Health & Family Welfare, Janakpuri, New Delhi-58. ##Department of Botany, Utkal University, Vani Bihar, Bhubaneswar, Orissa
A B S T R A C T
Kakoli (Roscea procera Wall.) of Family Zingiberaceae and Ksheerkakoli (Lilium polyphyllum D. Don) of family Liliaceae, are mentioned as drugs of Astavarga, but the botanical identity of both the species are controversial and different drugs are sold/ used in the name of Kakoli and Ksheerkakoli. This has created frightening problem with
made to lay down pharmacopoeial characters and to identify botanical identity of market samples by comparision with genuine drugs.
Key words: Macroscopy, Microscopy, Thin Layer Chromatography, physico chemical evaluation.
INTRODUCTION
The identity of Astavarga drugs suffer a lot of confusion in Ayurvedic literature. According to various Ayurvedic Nighantu books composed or commented by different Vaidyas, it constitutes a group of eight drugs, which form an important constituent of a number of Ayurvedic preparations. These are known in Sanskrit as Jivaka, Rishibhak, Mahameda, Meda, Kakoli, Ksheerkakoli, Riddhi and Vriddhi.[1-3]
Bhav Mishra in his Bhav Prakash Nighantu has further mentioned that none of Astavarga drugs are true.[4] Kakoli and Ksheerkakoli have been described under “Brhneeya” (the drugs which promote the formation of mansadhatu
[5] Caraka Samhita,[6] Sushruta Samhita[7] and Astanghridaya.[8] Till date none has tried to work out on their pharmacognosy and chemistry evaluation therefore they need further investigation. There is however a number of crude drugs
Chinmay, et al.: Pharmacognostical & Phytochemical Studies of Roscea procera (Kakoli) and Lilium Polyphyllum (Ksheerkakoli)
covered with hard membranous scales arranged in a concentric manner and breaking readily with a short fracture; cut surface white to creamish-yellow and starchy; scars of adventitious roots seen; odour, pleasant; taste, bitter.
MicroscopicTransverse section of bulb shows concentric layers of scale leaves; axis of bulb show three concentric layers of scale leaves. T.S of scale shows with an outer and inner epidermis consisting of single layered parenchymatous, pentagonal cells with mucilage; cuticle of both epidermis
of all the samples were carried out adopting standard procedures.[11-13] The thin layer chromatography of 90 per cent ethanolic extract of all samples were performed on pre-coated silica gel 60 F254 aluminium plates and the plates were developed in solvent system Toluene: Ethyl acetate: Methanol (8.0:1.6:0.4). The developed plates were observed under UV 254 nm and UV 366 nm and after derivatization visualisation under UV 366 nm.
Market samples RP1, RP2, RP3 and LP1, LP2, LP3 were collected in the name of Kakoli and Ksheerkakoli from New Delhi, Jaipur (Rajasthan), Mandi (Himachal Pradesh), respectively and compared pharmacognostically as well as phytochemically with authentic drugs.
RESULTS
Macro – and microscopical studies
of Roscea procera Wall.
MacroscopicRoots occur in bunches of 4-15; straight or curved, dark brown; each root about 3-8 (10) cm long, upto 0.9 cm thick; external surface rough due to presence of longitudinal wrinkles; odour, slightly aromatic; taste acrid.
MicroscopicTuberous root shows circular in outline; cork 10-12 layered, consisting of thin-walled, tangentially elongated, almost
content; below the cork phellogen layer is present; cortex consisting of oval to elongated, thinwalled, parenchymatous
of usual elements, xylem vessels arranged alternatively with phloem patches, vessels mostly solitary with spiral thickening
and xylem parenchyma are associated with xylem vessels; phloem consists of sieve tubes, companion cells, phloem parenchyma; pith composed of oval to polygonal, thin-walled, parenchymatous cells.
PowderGreenish-yellow; slightly aromatic in smell; shows cork cells in surface view and section view; in sectional view cork
crystalline material, simple, ovoid to ellipsoidal starch grains,
Macro - and microscopical studies of Lilium polyphyllum D. Don
MacroscopicWhole bulbs (tuber), conical 1.4 to 3.0 in width and 2.5 to 4.0 cm in length, transluscent with slight longitudinal ridges,
Figure 1a: T.S. root of Roscea procera Wall. (diagrammatic); Figure 1b: T.S. root of Roscea procera Wall. (showing cork and cortex region); Figure 1c: T.S. root of Roscea procera Wall. (showing phloem and xylem woody region); Figure 1d: Powder characteristics of Roscea procera Wall. (diagrammatic)RETRACTED
Chinmay, et al.: Pharmacognostical & Phytochemical Studies of Roscea procera (Kakoli) and Lilium Polyphyllum (Ksheerkakoli)
The detail macro- and microscopic characters of all samples RP1-RP3 and LP1-LP3 were studied carefully and compared with authentic samples of Roscea procera Wall. and it was found that only RP1 are genuine samples and the macro- and microscopical characters of RP2 resembles with Withania somnifera (L.) Dunal. The details of pharmacognostical characters are given below
MacroscopicRoot tuberous attaining diameter of 1 to 2.5 cm, occasionally branched, the distal ends taper slightly woody, outer surface is yellowish brown in colour, smooth but sometimes shallow
tuberous root reveals an inner smooth white starchy tissue surrounded by an outer narrow brown ring.
MicroscopicTransverse section of root shows narrow cork, a moderate cortex and a large wood. Cork shows 6 to 8 rows of thin walled cubical to slightly tangentially elongated cells. The inner portion of cork tissue. Cortex is 3 to 4 mm in thickness, composed of 14-18 thin walled slightly tangentially elongated cells. Simple to compound type of starch grains are abundantly embedded in cortex and vary in size, compound starch grains having 2-5 components
is slightly wavy and horny, mesophyll consists of 6 to 9 layered hexagonal parenchymatous cells; starch grains gelatinized which are eccentric type (hilium present on one side), deposited in parenchymatous cells and are of various shape i.e. polygonal, oval and truncated and 6-10 in one parenchymatous cell. Raphides ranging
the mesophyll; surface view of upper epidermis show compactly arranged rectangular, elongated thin walled cells. Longitudinal section of bulb scale shows tracheids, which is composed of narrow, elongated and tubular
secondary cell wall material is deposited on primary cell wall forming a ladder like pattern.
Transverse section of root shows single layered epidermis, the cells of epidermis are pentagonal in shape. Cortex is made up of parenchymatous cells which are hexagonal in shape, without intercellular spaces. Below the cortex is a thick walled single layered endodermis. Pericycle is present which is composed of thin and single layered cells. Primary xylem is distributed towards the pith zone and surrounded by phloem. The main part of pith area is occupied by metaxylem. Pith is parenchymatous.
PowderPowder creamish with pleasant smell; raphides present; powder treated with ruthenium red, mucilage turns bright pink.
Figure 2a: Bulb peel of Lilium pophyllum D.Don; Figure 2b: T.S. scale of Lilium pophyllum D.Don; Figure 2c: L.S. scale of Lilium pophyllum D. Don; Figure 2d: T.S. root of Lilium pophyllum D.Don
Chinmay, et al.: Pharmacognostical & Phytochemical Studies of Roscea procera (Kakoli) and Lilium Polyphyllum (Ksheerkakoli)
vessels; radially cut medullary rays crossing the vessels and
The physico-chemical analysis (Table 1) and TLC of 90 percent ethanolic extract of powders of all samples are carried out using solvent system Toluene: Ethyl acetate: Methanol (8.0:1.6:0.4). and observation are compared.
The pharmacognostic studies of LP1-LP3 were also carried out and observed that the macro-and microscopic characters of sample LP3 was found similar to Chlorophytum arundinaceum. The details of pharmacognostical characters are given below:-
MacroscopicPeeled dried tuberous root shriveled and sharply tapering at both ends cylindrical and arcuate or 5-7 cm long, 0.4 (0.7) to 1 (1.2) cm thick, longitudinally ridged and furrowed, transversely cracked at places; unpeeled tubers more rough, exhibit root scars. Fracture short and brittle; root brownish
passing inner to centre is enclosing the wood and intercepted by medullary rays. Phloem composed of small sized thin walled polygonal cells. Thin walled 2-3 layered broadly rectangular cambium tissue is also present. Xylem is a wide consisting of isolated or rarely groups of 2 to 3 vessels
of the wood, parenchyma are vesicentric and paratracheal and medullary rays are bent at places expecially when run adjacent to the vessel.
PowderShows abundant, simple and compound, spherical, oval or cup shaped starch grains with slit like or stellate hilum, scattered as such throughout or embedded in the parenchymatous cells of the cortex; microsphenoidal crystals of calcium oxalate embedded in the parenchymatous cells of the cortex; fragments of suberised cork in transverse and surface view; fragments of longitudinally cut and horse-shoe shaped pitted
Table 1: Observations of Physicochemical Parameters of Powdered Samples of Kakoli
Figure 3: TLC fingerprint of 90% ethanolic extract of samplesSamples of Roscoea procera fromRP 1: New Delhi, RP 2: Jaipur (Rajasthan), RP 3: Mandi (Himachal Pradesh), RP 4: Tarikhet (Uttarakhand)
RETRACTED
Chinmay, et al.: Pharmacognostical & Phytochemical Studies of Roscea procera (Kakoli) and Lilium Polyphyllum (Ksheerkakoli)
the apical and lower parts of the root. An endodermis and pericyclic layer are distinct. Groups of phloem tissue alternating with exarch xylem bundles in a ring encircled
of epiblema and exodermis tissue.
PowderPowder shows bundles or free acicular crystals of calcium oxalate scattered as such throughout and emebedded in
radially arranged cells of epiblema in surface view; fragments
and acicular crystals of calcium oxalate; pitted and reticulate
The physico-chemical analysis (Table 2) and TLC of 90 percent ethanolic extract of powders of all samples are
externally, whitish internally. Taste slightly sweet and mucilaginous and devoid of any odour.
MicroscopicDiagrammatic Transverse section of the root is somewhat circular in outline exhibiting epiblema, an outermost layer, followed by exodermis and wide parenchymatous cortex containing bundles of acicular crystals of calcium oxalate and central very narrow pith encircled by alternately arranged phloem and xylem bundles.
Detailed Transverse section of unpeeled root shows a layer
dome shaped cells bearing long hairs which usually get detached in the dry samples. Underneath this lie 3 to 5 rows of tangentially elongated cells, exodermis which are oval in shape and radially arranged followed by wide parenchymatous cortical zone containing mucilage and bundles of acicular crystals of calcium oxalate, more in
After derivatization visualisation under UV 366 nm (Rf values)
Chinmay, et al.: Pharmacognostical & Phytochemical Studies of Roscea procera (Kakoli) and Lilium Polyphyllum (Ksheerkakoli)
carried out using solvent system Toluene: Ethyl acetate: Methanol (8.0:1.6:0.4), and result compared to the authentic samples.
DISCUSSIONS AND CONCLUSION
Macroscopically the roots are straight and curved and dark brown in Roscea procera and in Withania somnifera roots are occasionally branched, buff to yellow brown while longitudinal wrinkles are present at outer surface in both species.
Microscopically the tuberous root of Roscea procera shows circular in outline, cork consisting of thin walled cells with reddish brown content, cortex thin walled with oval to ellipsoidal starch grains, 5-11 dia. And vascular bundles composed of usually elements, xylem vessels arranged alternatively with phloem parenchyma. The roots of Withania somnifera shows narrow cork, moderate cortex and a large wood. In cortical region starch grains are abundant. Starch grains are of 2 types, compound with
narrow ring of phloem passing inner to centre, enclosing the wood and intercepted by medullary rays are present.
and simple pitted vessel and tracheids in powder of Roscea procera and in Withania somnifera microspheroidal crystals of calcium oxalate present in cortical region and fragments of suberised cork in transverse and surface view and longitudinally cut and horse shoe shaped pitted vessels.
In Lilium polyphyllum bulbs are conical, translucent with slight longitudinal ridges covered with hard membranous scales. While peeled tuberous roots of shriveled, cyclindrical, sharply tapering at both ends.
Transverse section of bulb of Lilium polyphyllum shows a concentric rings of scale leaves. TS of root shows a single layered epidermis followed by parenchymatous cortex without intercellular spaces and the main part of pith is occupied by metaxylem. However in Chlorophytum arundinaceum outer layer of epiblema with long hairs which usually get detached in dry samples, followed by compact layers of exodermis and a wide parenchyma cortical zone with mucilage and bundles of acicular crystals of calcium oxalate.
The observation of physico-chemical evaluation indicates that the most drugs available in market are not genuine and adulterated with other plant drugs which are easily available in the market. In this dimension pharmacognostic anjd phytochemical studies on market samples is a substantial
Figure 5a: T.S. root of Withania somnifera (L.) Dunal (diagrammatic); Figure 5b: T.S. root of Withania somnifera (L.) Dunal (showing cellular details); Figure 5c: Powder characteristics of Withania somnifera (L.) Dunal
RETRACTED
Chinmay, et al.: Pharmacognostical & Phytochemical Studies of Roscea procera (Kakoli) and Lilium Polyphyllum (Ksheerkakoli)
12. Chase CR, Pratt FJ. Fluorescence of powdered vegetable drugs with particular reference to development of a system of identification. J Am Pharm Assoc1949; 38:324-3.
13. Anonymous. Indian Pharmacopoeia, 2nd ed., Delhi; 1966.
step and it further requires a long term study and this type
material and for Industries also.
REFERENCES
1. Puri HS. Traditional Herbal Medicine for Modern Times, Rasayana – Ayurvedic Herbs for Longevity and Rejuvenation. London: Taylor & Francis; 2003
2. Pandey G. Dravyaguna Vijnana. IInd edition: Varanasi India. Krishnadas Academy; 2002.
3. Gogte Vaidya V M. Ayurvedic Pharmacology and Therapeutic Uses of Medicinal Plants – Dravyagunavignyan. (English Translation). Mumbai: Bharatiya Vidya Bhavan; 2000.
Figure 6a: T.S. root of Chlorophytum arundinaceum Baker (diagrammatic); Figure 6b: T.S. root of Chlorophytum arundinaceum Baker (showing tissues of outer region); Figure 6c: T.S. root of Chlorophytum arundinaceum Baker (showing tissues of central region); Figure 6d: Powder characteristics of Chlorophytum arundinaceum Baker
