STUDIES ON MORPHOLOGY AND MEDICINAL EFFICACY OF NANOPARTICLES SYNTHESIZED FROM TWO MEDICINALLY IMPORTANT CURCUMA SPP.,

Turmeric,a spice and medicinal plant, has proven to be the most effective nutritional supplement for the human body. The present study made an attempt to compare the morphological and anatomical characterization of two species of Curcuma, to standardize a user eco-friendly protocol for the biosynthesis of silver nanoparticles (AgNPs) through biological method, and to evaluate the antifungal activity of these species. C longa had curcumin cells while C.caesia had no curcumin cells, only starch grains. Ag- nanoparticles were synthesized at a much faster rate in C.longa , when compared to that in C.caesia . Antifungal activity is faster for AgNO 3 mixture of C.longa , and much slower in aqueous extract of C.longa and C.caesia and AgNO 3 mixture of C.caesia. The fungal colonies were destroyed soon after the colony was inoculated with AgNO 3 mixture of C.longa , but no visible change in diameter occurred in the fungal colony after the inoculation of other mixtures. Pellets could be made from powdered rhizomes of both species which could be used as a safe alternative to preventive diseases.

Turmeric,a spice and medicinal plant, has proven to be the most effective nutritional supplement for the human body. The present study made an attempt to compare the morphological and anatomical characterization of two species of Curcuma, to standardize a user ecofriendly protocol for the biosynthesis of silver nanoparticles (AgNPs) through biological method, and to evaluate the antifungal activity of these species. C longa had curcumin cells while C.caesia had no curcumin cells, only starch grains. Ag-nanoparticles were synthesized at a much faster rate in C.longa, when compared to that in C.caesia. Antifungal activity is faster for AgNO 3 mixture of C.longa, and much slower in aqueous extract of C.longa and C.caesia and AgNO 3 mixture of C.caesia. The fungal colonies were destroyed soon after the colony was inoculated with AgNO 3 mixture of C.longa, but no visible change in diameter occurred in the fungal colony after the inoculation of other mixtures. Pellets could be made from powdered rhizomes of both species which could be used as a safe alternative to preventive diseases.

…………………………………………………………………………………………………….... Introduction:-
Medicinal plants play an important role in the development of potent therapeutic agents. Herbal medicines are currently in demand and their popularity is increasing day by day hence it plays a vital role for the development of new drugs. Herbal medicines have a vital role in the prevention and treatment of various diseases. Man since time immemorial has been using plants or natural product as medicine to promote and maintain good health (Parrota, 2001). Over the past few decades, there has been increased reports on the biological activities of turmeric, along with their medicinal uses (Sarker and Nahar, 2007) Curcuma longa and C. caesia are two medicinally important species belonging to the family Zingiberaceae andCurcumin is an important phytochemical seen in the members of the genus Curcuma.
Recently, attention has been paid to utilize eco-friendly and bio-friendly plant based products for the prevention and cure of different human diseases. Most of the people have faith in traditional medicine, particularly plant drugs for their primary health care. Even though these plants are commonly available, the multiple uses of these species have increased its commercial demand, resulting in over-exploitation. The use of nanotechnology for 'phytotherapy' or treatment of various diseases by herbal medicines, including herbal drug delivery where current and emerging nanotechnologies could enable entirely novel classes of therapeutics has been reported. Interestingly, pharmaceutical sciences are using nanoparticles to reduce toxicity and side effects of drugs. The ISSN: 2320-5407 Int. J. Adv. Res. 6(10), 930-936 931 biologically synthesized nanoparticles with plant products have better chemotherapeutic effects against microbial diseases. Turmeric is commercially available in different forms along with enhancers like bromelain.
The present study made an attempt to compare the morphological and anatomical characterization of two species of Curcuma, to standardize a user eco-friendly protocol for the biosynthesis of silver nanoparticles (AgNPs) through biological method, and to evaluate the antifungal activity of these species

Morphological characterization
The morphological characters that were observed were scale leaf length, width of scale leaf, width of clump, length of fingerling ,nodal length, intermodal length and color of rhizome

Statistical analysis
Ten observations were taken and considered to calculate the standard deviation of all the morphological characters..

Anatomical studies
Rhizomes of C.caesia and C.longa and their available tubers were hand sectioned .The sections were stained using safranine, mounted on a slide and viewed under microscope.
The cortical and stellar characters like epidermal hair, number of epidermal layers, vascular bundles in outer and inner cortex and the curcumin content in both species.

Synthesis of nanoparticles
Aqueous extracts of C.caesia and C.longa, 2ml each are taken in separate test tubes. One test solution of both C.caesia and C.longa are kept as control.10ml AgNO 3 is added to the Aqueous extracts of C.caesia and C.longa separately, the mouth of testtube is covered using silverfoil and left unmdisturbed.The color change is noted periodically.

UV-vis spectroscopy
The absorbance of AgNO 3 mixed solutions is found out with the help of UV-vis spectrometer. The solution is transferred into cuvette and then absorbance is calculated.

Anti fungal activity
Aqueous extracts and AgNO 3 mixtures of C.caesia and C.longa are inoculated into separate fungal colonies to find out their antifungal activity.

Pellatisation
The shade dried rhizomes of C.caesia and C.longa are powdered and pellatised using a hydraulic press.

Results and Discussion:-
The present work is a comparative study on morphology and medicinal efficacy of nanoparticles synthesized from two medicinally important Curcuma species C. longa and C.caesia. The present studuy revealed the presence of medically active constituents.

Morphological characterization:-
The freshly removed clumps of C.caesia and C.longa were used for morphological characterization. Morphological characters of both species showed variations, as in Table 1.  Anatomical characterization : The fingerlings of C.caesia and C.longa of similar size were selected for hand sectioning and stained and viewed under microscope Anatomical characters of both species showed extreme variations as depicted in Table 2. Curcumin cells were found in plenty in C.longa but were totally absent in C.caesia (Fig 1b, 2b). Both schizogenous type (secretory duct originating in the intercalary meristem) and lysigenous type (formation in meristematic and mature parts of rhizomes) have been reported in C.longa (Ravindran et   Nanoparticle Synthesis C longa, C.caesia extracts were mixed with 0.1 N AgNO 3 solution, and the change in colour was noted periodically. It was observed that when Curcuma longa ,Curcuma caesia extract is mixed with 0.1 N AgNO 3 solution, a deep reddish brown colour is obtained after 24 hours ( Table 3). The colour indicates the formation of Ag colloid by chemical reduction of AgNO 3.

UV-Spectroscopy
The UV-vis spectra recorded on the rhizome extracts mixed with AgNO 3 (3 mM) solutions. UV-vis absorption spectra have proved to be quite sensitive to the formation of silver colloids because silver nanoparticle exhibit an intense absorption peak due to surface excitation. The UV-vis spectra recorded on the rhizome extracts mixed with AgNO 3 (3mM) solution as a function of time of reaction. Immediately after addition of AgNO 3 solution to the extract, the colour of clear extract first changed to a ring formations of a reddish brown. At this stage, formation of metal nanoparticles due to reduction was observed. The generations of colour is due to excitations of surface plasmons in a metal nanoparticle. Followed by UV-vis spectroscopy, absorption of Ag-nanoparticles was maximum at 277..2 nm for C.caesia and 363.6nm for C longa, (Figs 3, 4, 5) which steadily increased in intensity without showing any shift of the wavelength maximum. This depicts the reductions of silver ion taking place at a faster rate and the completions of reaction. UV-vis absorption spectra have been proved to be quite sensitive to the formations of silver colloids because silver nanoparticle exhibit an intense absorption peak due to surface Plasmon. Fig. 3

Anti Fungal Activity
The aqueous and AgNO 3 extract of both C.caesia and C.longa were inoculated into fungal colonies grown in petridishes. AgNO 3 extract of yellow turmeric killed the fungal colonies immediately after inoculation (Fig 5 ).

Preparation of pellets from rhizomes
The rhizomes of both C.caesia and C.longa were taken, sliced separately and dried in shade. The dried specimens were powdered thoroughly and kept in two separate bottles. Powders are weighed, wrapped in a tissue paper and made into pellets. These pellets can be used as medicinal supplements.
The rhizomes of black turmeric and yellow turmeric with almost equal width are taken ,sliced separately and dried in shade. The dried specimens are powdered thoroughly and kept in two separate bottles.0.05g of black turmeric powder is weighed out . The same procedure is repeated in the case of yellow turmeric powder. The powdered form of specimen is changed to pellets form with the help of pellatizer A pressure of 3ton was applied during the preparation of each pellets.
The pellatizer or Hydraulic press works according to Pascal's Law 'In any fluid the static pressure is exerted on the walls of the container and in the fluid. These forces act perpendicular to the walls of the container.
The press is capable of producing maximum pressure upto 15 tons. It is compact in construction and occupies only a bench space. Acrylic screens are provided for the safety of the operator. The base plate has provision for bench mounting. They are used to prepare pellets. Pellets could be successfully made (Fig.7) from both species, indicating a viable method for preparation of consumable pellets for routine health care Antifungal activity of the essential oil of Curcuma caesia was experimented by Banerjee and Nigam (1976). The rhizomes were tested for antifungal activity against several human and plant pathogenic fungi. Dilutions of the oil in ethylene glycol were tested by an agar diffusion procedure on plates seeded with the test isolates. Some antifungal effect was noted, but no consideration was given in designing the experiment to the fact that oil diffused poorly through agar gels. The significance of the results therefore remains unclear. Antimicrobial efficacy of essential oil from Curcuma caesia was done by Garg and Jain, 1998. The essential oil from rhizome of C. caesia, rich in curcumene, ionone and turmerone, was tested for antimicrobial activity against Morphological characters indicated that C longa has larger scale leaf characters in terms of length and breadth, clump width, fingerling length and nodal length, while C.caesia has higher intermodal length. C longa had curcumin cells while C.caesia had no curcumin cells. Ag-nanoparticles were synthesized at a much faster rate in C.longa, when compared to that in C.caesia. Absortipon of Ag-nanoparticles was maximum at 277..2 nm for C.caesia and 363.6 nm for C longa. Antifungal activity is maximum for AgNO 3 mixture of C.longa, and absent in aqueous extract of C.longa and C.caesia and AgNO 3 mixture of C.caesia. The fungal colonies were destroyed soon after the colony was inoculated with AgNO 3 mixture of C.longa, but no visible change in diameter occurred in the fungal colony after the inoculation of other mixtures. Pellets could be made from powdered rhizomes of both species which could be used as a safe alternative to preventive diseases.