450 J SCI IND RES VOL 66 JUNE 2007 Journal of Scientific & Industrial Research Vol. 66, June 2007, pp. 450-456 Botanical standardization of three commercially important Curcuma species Sharad Kumar Srivastava*, Ajay Kumar Singh Rawat, Shanta Mehrotra and P Pushpangadan Pharmacognosy & Ethnopharmacology Division, National Botanical Research Institute, Lucknow 226 001 Received 24 November 2006; accepted 24 January 2007 Amraharidra, botanically equated to Curcuma amada (Family, Zingiberaceae), is an important drug of Ayurveda. Rhizomes of other species (C. aromatica and C. zedoaria) of this genus, being rich in essential oil contents, are sold in herbal drug markets by the same vernacular name. Present study is aimed to standardize botanically all these three species for their authenticity, identity and validity. Keywords: Amraharidra, Anatomy, Curcuma, Pharmacognosy Introduction Results Amraharidra, botanically equated to Curcuma On morphological studies, leaves of C. amada amada (Family, Zingiberaceae), is an important drug of (Fig. 1) were found sparsely hairy, while C. aromatica Ayurveda. It is considered to be haemostatic, stomachic (Fig. 2) leaves were profusely hairy on ventral side and and carminative. Its local application makes the skin soft C. zedoaria (Fig. 3) had purple midrib. Rhizome of all and reduces itching and dryness. It is also applied locally the three species can easily be differentiated on the basis on contusions and sprains1-3. Hence, traders of the of odour and taste. It is raw mango like with sweet taste country subject it to adulteration/substitution. Rhizomes in C. amada, while C. aromatica has camphoraceous of other species (C. aromatica and C. zedoaria) of this odour and bitter spicy taste and C. zedoaria possess genus, being rich in essential oil contents, are sold in strong aromatic camphoraceous odour and pungent taste. herbal drug markets by the same vernacular name. The colour of cut surface is creamy white, creamy yellow Except Srivastava et al4, no pharmacognostic details are and orange yellow in C. amada, C. aromatica and C. available on these three species. This study presents zedoaria respectively (Table 1). macro and microscopical characters undertaken to On microscopical examination, rod shaped differentiate three Curcuma species. starch grains and fibres are observed only in C. amada, while the shape and size of starch grains are variable in Materials and Methods remaining two species. Fibre content is absent in C. Plant material was collected from Palkkad, amada and C. zedoaria while it was present in C. Kerala region (Sharad, LWG 221248, 1999; Sharad, aromatica (Table 2). Length and width of the tracheids LWG 221249. 1999; Sharad, LWG 221250, 1999) and and vessels were higher in C. zedoaria (Table 3). 6-7 rhizomes were preserved in 70% ethyl alcohol for Behaviour of powdered drug with different chemical histological studies. Microtome sections were cut (TS reagents was also studied (Table 4). & TLS) and stained with safranin and fast green and photographed with Nikon F70X camera5. Conclusions Starch grains and presence/absence of oil containing cells, fibers and some tracheiary elements *Address for correspondence can be utilized as identifying marker for these species. E-mail: [email protected] SRIVASTAVA et al.: BOTANICAL STANDARDIZATION OF THREE CURCUMA SPECIES 451 Fig. 1 — Macroscopic and microscopic characters of C. amada rhizome 452 J SCI IND RES VOL 66 JUNE 2007 Fig. 2 — Macroscopic and microscopic characters of C. aromatica rhizome SRIVASTAVA et al.: BOTANICAL STANDARDIZATION OF THREE CURCUMA SPECIES 453 Fig. 3 — Macroscopic and microscopic characters of C. zedoaria rhizome 454 J SCI IND RES VOL 66 JUNE 2007 Table 1— Comparative macroscopic characters of three Curcuma species Characters Curcuma amada Curcuma aromatica Curcuma zedoaria Leaf Leaves hairy on ventral Leaves more hairy on Leaf mid-rib purple side ventral side Rhizome Profusely branched, buff Less branched, lateral Less branched, lateral coloured, bulbous with branches finger shaped and branches are cylindrical short & smooth fracture, palmately attached, root tubers present root tuber absent root tubers present Odour Raw mango like smell Sweet camphoraceous smell Strong aromatic and sweet in taste and bitter spicy taste camphoraceous smell with acrid or pungent taste Colour of Creamy white Creamy yellow Orange yellow rhizome Flowering June-August May-June April-July Table 2 — Comparative microscopic characters of three Curcuma species Characters Curcuma amada Curcuma aromatica Curcuma zedoaria Epidermal hairs Unicellular, thick 1-2 celled, branched, thick 2 celled, thick walled, lignified walled, lignified walled, lignified with swollen base Periderm 4-7 layered 3-5 layered 2-5 layered Outer cortex Very few collateral Lesser number of scattered More than 25 collateral, scattered vascular collateral vascular bundles, scattered vascular bundles, bundles, starch grains starch grains present full of starch grains occupied maximum area Inner cortex Amphicribal vascular Amphicribal vascular Amphicribal vascular bundles bundles arranged in bundles arranged in patches arranged in distinct zones distinct zones Vascular Trachieds with spiral Trachieds with spiral and Tracheids with spiral and elements and scalary and vessels scalary and vessels with scalary and vessels with with reticulate reticulate thickening, reticulate thickening, thickening, bundle sheath absent, bundle sheath absent, bundle sheath present, fibre present very few fibre absent fibre absent Lysigenous oil Less in number Present Present cells Curcumin cells Very few numbers Uniformly present in both More in inner zone than outer present the zones but smaller in size zone as compared to C. zedoaria Starch grains Mostly rod shaped Variable in shape and Variable in shape and number. number Table 3 — Comparative maceration study of three Curcuma species Elements Curcuma amada Curcuma aromatica Curcuma zedoaria Mean SD Mean SD Mean SD Trachieds Length 432.635 ±13.854 420.719 ±8.937 643.453 ±0.861 Width 13.290 ±0.158 14.207 ±0.283 15.123 ±0.241 Vessels Length 737.863 ±8.684 670.777 ±15.388 744.839 ±0.687 Width 14.665 ±0.210 15.397 ±0.252 15.829 ±0.261 Fibres Length - - 658.118 ±2.863 - - Width - - 9.166 ±0.000 - - Table 4 — Fluorescence analysis of the rhizome of three Curcuma species Sl No. Treatement Curcuma amada Curcuma aromatica Curcuma zedoaria Day light UV-254 nm. UV-366 nm. Day light UV-254 nm UV-366 nm Day light UV-254 nm UV-366 nm 1 Powder (P) as such Light brown Green Fluorescent light Light brown Light brown Black Yellowish brown Brown Brown SRIV green 2 P.+ nitrocellulose Light orangish Dark green Fluorescent brown Dark brown Brown Black Fl. yellow Fl. yellow Yellow AST in amyl acetate brown AVA AVA 3 P.+ 1N NaOH Light brown Green with Green Dark brown Brown Black Brick red Dark brown Brown in water with yellow tinge yellowish tinge et al. 4 P.+ 1N NaOH + Yellowish green Dark green Yellowish green Dark brown Brown with Black Brick red Brown with Black : BOT nitrocellulose with greenish tinge greenish tinge yellowish tinge in acetate ANICAL 5 P.+ 1N HCl + Orange Dark green Light green Dark brown Brown with Black Fl. yellow Fl. yellow Yellow nitrocellulose with greenish tinge greenish tinge in amyl acetate ST 6 P.+ 1N NaOH Light brown Light green Yellow with purple Dark brown Dark brown Black Brick red Brown with Black ANDARDIZA in methanol with orange tinge margins yellowish tinge 7 P.+ 50% KOH Cream Green No change Dark brown Dark brown Black Brick red Brown with Black yellowish tinge 8 P. + 1N HCl No change No change No change Light brown Light brown Black Brown Light brown Black TION OF 9 P.+50% H2SO4 Brownish green Bright green Green Brown Light brown Black Black Dark brown Black tinge with red tinge 10 P.+50% HNO3 Light brown No change No change Yellow Brown with Black Brown with Brown with Brown with with red tinge greenish tinge yellowish tinge greenish tinge violet tinge THREE CURCUMA 11 P.+ conc. HNO3 Brownish yellow Greenish yellow Brown Yellow with red Brown with Black Muddy yellow Brown with Black tinge greenish tinge greenish tinge 12 P.+ acetic acid Yellow Bluish green Yellow Light brown Light brown Black Light brown Light brown Black 13 P.+ conc. H2SO4 Brown Bright green Bright green Black Black Black Black Black Black SPECIES 455 456 J SCI IND RES VOL 66 JUNE 2007 Acknowledgements 2 Chopra R N, Chopra I C, Handa K L & Kapoor L D, Authors thank Director NBRI for guidance and Indigenous Drugs of India (U N Dhur and Sons, Kolkata) 1958, 503-505. providing all the facilities to conduct this work, and also 3 The wealth of India, vol 2B (Publication and Information thank Dr K Sahai and Mr Ramsamujh for technical Directorate, CSIR, New Delhi) 1988, 114-118. assistance. 4 Srivastava, S K, Khatoon, S, Rawat, A K S & Mehrotra, S, Pharmacognostical studies of rhizomes of Amraharidra, J Sci Nomenclature Ind Res, 61 (2002) 17-19. ICO, Inner cortex; CK, Cork cells; IVB, Inner vascular bundle; OVB, 5 Josansen D A, Plant Microtechnique (McGraw Hill Book Co. Outer vascular bundle; CO, Cortex; FR, Fiber; ST, Starch; VS, Inc., New York and London) 1940, 102-104. Vessels; XY, Xylem; ED, Endodermis; PR, Pericycle; HR, Hairs; 6 Chase C R & Pratt R J, Fluorescence of powdered vegetable TC, Tannin containing cells drugs with particular reference to development of a system of identification, J Am Pharm Assoc, 38 (1949) 324-331. References 7 Kokoski J, Kokoski R & Slama F J, Fluorescence of powdered 1 Kirtikar K R & Basu B D, Indian Medicinal Plants (L M Basu vegetable drugs under ultraviolet radiation, J Am Pharm Assoc, Publication, Allahabad) 1933, 2422-2423. 47 (1958) 75-77..