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TURMERIC

INTRODUCTION

Man has been using herbs and plant products for combating diseases since times immemorial.
The Indian subcontinent is enriched by a variety of flora both aromatic and medicinal plants.
This extensive flora has been greatly utilised as a source of many drugs in the Indian
traditional system of medicine. In India, the earliest mention of the use of medicinal plants is
to be found in Rigveda which was written between 4500-1600 1. Turmeric (Haridra) is one
such medicinal plant explained extensively in Indian material medica (Dravyaguna Sastra). It
is an auspicious beauty spot, daily applied on the forehead by Hindu females. Application of
turmeric a paste to the bride is an essential procedure of Hindu rituals 2. In Ayurveda, turmeric
has been well documented for its therapeutic potentials and described in Dashemani
Lekhaniya (emaciating), Kusthagna (Anti-dermatosis), Visaghna (Anti-poisonous)3.

HISTORY OF TURMERIC

Haridra in Sanskit means an efficacious drug for jaundice4. It is known to be one of the
oldest spices that have been used in Western and Southern parts of India for thousands of
years and is a major part of Ayurvedic medicine5. Turmeric has been used in Asia for
thousands of years and is a major part of Siddha medicine 6. It was first used as a dye and then
later for its medicinal properties7. The origin of the name is uncertain, possibly deriving from
Middle English/Modern English as turmeryte or tarmaret. Speculation exists that it may be of
Latin origin, terra merita (Merited earth)8.

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REVIEW OF LITERATURE- PHYTOCHEMISRTY & PHARMACOLOGICAL
INVESTIGATIONS

Molecular Consituents in Turmeric- Turmeric has hundreds of molecular constituents, each


with a variety of biological activities. For instance, there are at least 20 molecules that are
antibiotic, 14 are known cancer preventives, 12 that are anti-tumor, 12 are anti-
inflammatory and there are at least 10 different anti-oxidants. Infect, 326 biological
activities of turmeric are known. This is also testimony to the use of whole herbs and not just
isolated molecules. Speaking of molecules by far the most researcher in turmeric are the three
gold-coloured alkaloids curcuminoids viz. Curcumin, Demethoxycuccumin and
Bisdemethoxycurcumin. Most of the research done is with 95% Curcuminoids extract of
turmeric, through in its raw state turmeric is only 3-5% Curcuminoids. The yield of essential
oil in various parts is 1.3% in leaf, 0.3% in flower, 4.3% in root and 3.8% in rhizome. The
composition of essential oils9 obtained from root, rhizome, leaf and flower and nutritional
composition of Curcuma longa are given in Table-1.

Table 1: Composition of essential oils of Curcuma longa9

S.No Components Root (%) Rhizome (%) Leaf (%) Flower (%)

1 -Bisbolene 2.3 1.3 1.3 0.9

2 1,8-cineole 0.7 2.4 6.5 4.1

3 -Cymene 3.3 3.0 5.9 1.6

4 Curlone 0.6 10.6 0.2 0.3

5 Myrcene 0.1 0.1 2.3 0.2

6 -Pinene 0.1 0.1 2.1 0.4

7 Turmerone Not Mentioned 10.0 0.9 1.0

8 Terpinolene 0.1 0.3 26.0 7.4


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9 Curcumin 7.0 6.3 0.2 1.9

10 Dehydrocurcumin 4.3 2.2 Not Mentioned Not Mentioned

Table 2: Nutritional Composition of Turmeric (Curcuma longa)10

S.No Consituents Quantity per 100g

1 Ascorbic acid (mg) 50.0

2 Ash (g) 6.8

3 Calcium (g) 0.2

4 Carbohydrate (g) 69.9

5 Fat (g) 8.9

6 Food energy (K Cal) 390.0

7 Iron (g) 47.5

8 Niacin (mg) 4.8

9 Potassium (mg) 200.0

10 Phosphorus (mg) 260.0

11 Protein (g) 8.5

12 Riboflavin (mg) 0.19

13 Sodium (mg) 30.0

14 Thiamine (mg) 0.09

Pharmacological Investigation: Turmeric and its constituents play an important role in our
life. Which are given below:
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Turmeric has been found to have a hepatoprotective characteristic similar to that of
silymarin.

The volatile oils and curcumin and turmeric exhibit potent anti-inflammatory effects.

Turmeric and curcumin are also capable of suppressing the activity of several
common mutagens and carcinogens in a variety of cell types in both in vivo and in
vitro studies.

Turmeric extract and the essential oil inhibit the growth of a variety of bacteria,
parasites and pathogenic fungi.

Turmerics protective effects on the cardiovascular system include lower cholesterol


and triglyceride level, decreasing susceptibility of low density lipoprotein (LDL) to
lipid peroxidation and inhibiting platelet aggregation.

Constituents of turmeric exert several protective effects on the gastrointestinal tract.

Turmeric oil exhibited potent anti-trypsin and anti-hyaluronidase activity.

Constituents of turmeric affect Alzheimers disease.

Extract of turmeric suppresses symptoms associated with arthritis.

Turmeric and its extract inhibit angiogenesis.

Turmeric constituents can induce radioprotection.

Turmeric constituents inhibit proliferation of vascular smooth muscle cell.

Turmeric lower serum cholesterol levels.

Constituents of turmeric block the replication of HIV.

Turmeric constituents stimulate muscle regeneration.

Turmeric enhances wound healing.

Turmeric extract reduces the incidence of cholesterol gall bladder stone formation.

Turmeric constituents protects against cataract formation in lenses.

Turmeric protects against pancreatitis.

Turmeric extract corrects cystic fibrosis defects.

Turmeric suppresses the induction of adhesion molecules.

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Turmeric constituents inhibit androgen receptor and androgen receptor related
cofactor.

Constitutions of turmeric inhibit farnesyl protein transferase (FTPase).

Turmeric constituents inhibit scarring.

Turmeric oil containing turmerones exhibited a potent antioxidant activity in -


carotene.

Turmeric volatile oils suppress acute oedema10.

Stages in Tumor Progression inhibited by Curcumin11

EXPERIMENTAL WORK

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Collection and Identification of Plant Material

Pharmacognostic evaluation

Total Ash Value

Acid Insoluble Value

Water Soluble Ash

Determination of solvent extractive value

Loss on drying (LOD)

Preparation of the Extract- Ethanolic and Aqueous Extraction

Qualitative Chemical Test

Microscopical Studies- Morphological Study, Powder Charcterstics, Tranverse


Section Study, Fluorescence analysis, Micrometery and Physiochemical Parameters.

Market Sarvey for Standardized Ayurvedic fomulation of Curcuma longa

MATERIAL AND METHODS

Collection and Identification of Plant Material: The plant material was collected from the
market, Sundar Nagar, H.P. The organoleptic characters of fresh rhizomes and dried rhizome
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powder like colour, odour and taste and the macroscopic characters like size, shape, surface,
fracture were evaluated as per standard WHO guidelines.

PHARMACOGNOSTIC EVALUATION

Total ash value

It depicts the total amount of material produced after the complete incineration of the ground
drug above 400C to remove all the carbon atoms. 2g of powdered drug was weighed and
placed in the crucible and heated at about 400C. The crucible was cooled and the % of the
total ash with reference to the air-dried sample of the crude drug was calculated12.

Acid insoluble ash

Total ash obtained was dissolved in 1N HCl solution and heated for 5 min. The insoluble
matter was filtered in whatman filter paper; the filter paper was further dried at 70C and then
cooled. The residue was weighed and the percentage of insoluble ash of the crude drug w.r.t.
the air dried sample of crude drug was calculated12.

Water soluble ash

To the total ash crucible, 25ml double distilled water was added and boiled for about 5min.
Insoluble matter was collected on an ash less filter paper in a crucible, washed with hot water
and ignited for about 15min above 45C. The weight of the residue is subtracted from the
weight of the total ash. Content of water soluble ash in mg/g of the air dried material was
calculated12.

Determination of solvent extractive value

2g of the air- dried coarsely powdered drug was macerated with 100ml of different solvents
in closed flasks for 24h, and are shaken frequently. The solvent was filtered and the filtrate
was weighed in a Petri dish. The dish was evaporated on a water bath and then dried in an
oven at 100C. The dish was cooled and extractible value was calculated as % (w/w) with
reference to air dried drug.

Loss on drying (LOD)

LOD is the loss in weight in % (w/w) resulting from water and volatile matter of any kind
that can be driven off under specified conditions. 1g sample is transferred to a shallow bottle

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and weighed. Sample was distributed evenly and dried in a hot air oven at 105C for 1h with
the stopper open. After 1h, the stopper was closed and cooled at room temperature and the
bottle was weighed.

Preparation of the extract

The rhizomes of Curcuma longa were collected and dried in sun for 3 days, cut into small
pieces and again dried. The upper bark of the rhizome was removed to obtain the fresh
rhizome. The dried rhizome was then grinded to obtain a fine powder. The powder was again
dried and was ready for use.

Ethanolic extraction: The grinded powder was extracted with 500ml of dehydrated
ethanol and 1000ml double distilled water respectively by Soxhlation for 72h. The
extract was concentrated at temperature <45C. The residue was dried and
refrigerated.

Aqueous extraction: The grinded powder was then extracted with 1000ml double
distilled water containing 3 or 4 drops of chloroform for 48h. The extract was then
concentrated at temperature less than 45C.The residue was then dried and
refrigerated.

Qualitative chemical tests

Test for alkaloids13

(a) Mayer's test: To 2ml test solution, 2N HCl was added. The aqueous layer formed was
decanted and Mayer's reagent was added to it. A cream coloured precipitate indicates the
presence of alkaloids.

(b) Dragendroff's test: To 2ml test solution, and Dragendroff's reagent was added to it. A
reddish brown precipitate indicates the presence of alkaloids.

(c) Wagner's test: To 2ml test solution, and Wagner's reagent was added to it. A reddish
brown precipitate indicates the presence of alkaloids.

(d) Hager's test: To 2ml test solution, and Hager's reagent was added to it. A yellow coloured
precipitate indicates the presence of alkaloids.

Test for glycosides

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(a) To 2ml test solution, equal quantity of Fehling's solution A and B was added and solution
was heated. A brick red precipitate indicates the presence of glycosides.

(b) Legal's test: To 2ml test solution, pyridine and alkaline sodium nitroprusside was added
to obtain a blood red colour.

Test for flavonoids

(a) Shinoda test: To 2ml test solution, few fragments of Magnesium ribbon were added and
to it conc. H2SO4 was added drop wise. Pink scarlet or crimson red colour appears.

(b) Zinc chloride reduction test: To 2ml test solution, a mixture of zinc dust and conc. HCl
was added. A red colour is obtained after few minutes.

(c) Alkaline reagent test: To 2ml test solution, sodium hydroxide solution was added to give
a yellow or red colour.

Test for tannins

(a) Gelatin test: To 2ml test solution, 1% Gelatin solution containing 10% sodium chloride
was added to obtain a white precipitate.

(b) Ferric chloride test: To 2ml test solution, ferric chloride was added to give a blue green
colour.

Test for proteins and amino acids

(a) Millon's test: To 2ml test solution, Millon's reagent is added which gives a white
precipitate, which on heating changes to red.

(b) Ninhydrin test: To 2ml test solution, ninhydrin solution was added and the solution was
boiled. Amino acids and proteins when boiled with 0.2% ninhydrin reagent show a violet
colour.

Test for fats and fixed oils

(a) Stain test: Small amount of the extract was pressed between two filter papers; the stain on
the filter paper indicates the presence of fixed oils.

(b) Saponification test: Few drops of 0.5N alcoholic potassium hydroxide was added in
small quantity to the extract solution with a drop of phenolphthalein and heated on a water
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bath for 1-2h. The formation of soap or partial neutralization for the alkali indicates the
presence of fats and fixed oils.

Test for Sterols14

Liebermann-Burchard test: To the test solution, 3-4 drops of acetic anhydride was added,
the solution was boiled cooled and conc. Sulphuric acid (3 drops) was added. A brown ring
appears at the junction of the two layers. The upper layer turns green showing the presence of
steroids.

Test for triterpenoids15

Salkowski test: To the test solution 2ml chloroform was added with few drops of conc.
Sulphuric acid (3ml), and shaken well. Appearance of reddish brown colour at lower layer
indicates presence of steroids and that of yellow colour shows the presence of triterpenoids.

MICROSCOPICAL STUDIES

Fresh rhizomes of curcuma longa were subjected for the microscopical studies. The the
sections were cut by free hand sectioning. The numerous temporary and permenant mounts of
the microscopical section of the spicemen were made andexamined microscopically.
Photomicrophs of the microscopical section were taken with the help of MOTIC Digital
Microscope, provided with MOTIC IMAGE PLUS 2.0 software16,17,18.

Powder characteristics: Preliminary examination and behavior of the powder with different
chemical reagents was carried out as per reported method19,20.

Micrometry: Quantitative microscopy of the transverse sections and rhizome powder were
performed to determine the size and dimension of tissues, cell and cell content21,22.

Fluorescence analysis: Dried rhizomes were powdered and observed under visible light,
short ultra violet light, long ultra violet light after treatment with different reagents like
chloroform, ethyl acetate, methanol, petroleum ether, 50% sulphuric acid, 50% hydrochloric
acid, 50% nitric acid, 10% sodium hydroxide etc23.

RESULTS

Morphological Description: The central or primary rhizomes are ovate, cylindrical or


fusiform, curved, sometimes slightly branched into a Y-shape, 4-5 cm long, 5-30 mm in
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diameter, rough with wrinkled striations, distinct cyclic nodes and rounded scars of root
branches and rootlets. The organoleptic evaluation of the rhizomes revealed that the rhizomes
were Yellowish to yellowish-brown in colour, with characteristic and aromatic odour and
slightly bitter and pungent in taste. The results of morphological characters are mentioned in
Table no.1.

S.No Charaters Observations

1. Colour Yellow or Yellowish Brown

2. Odour Aromatic and Charactestics

3. Taste Slightly Bitter

4. Size 5-30mm in diameter

5. Length 4-5cm in length

6. Shape Finger shape

7. Surface Smooth and slightly rough

8. Texture Hard and Heavy

9. Fracture Short

Microscopical Description: The transverse section of the rhizome shows cork as an outer
layer followed by epidermis, cortex, and endodermis and ground tissue. Cork composed of
thin walled brown cells which is large and polygonal in shape. (Fig No. 1 & 2).

K- Cork Cell; P- Parenchyma filled


with starch paste

H- Oil Cell

G- Vessel.

Cork cells in surface view. X160


Epidermis is consist of thin
walled cubical cells of various dimension. The cortex consist of thin walled rounded
parenchymatous cells and having olioresin cells. These cells are filled with gelatinized starch
grains and yellow colouring matter. The ground tisse is parenchymatous and cells filled with
gelatinized starch grains and yellow pigment. Fibrovascular bundle and oil cells scattered
throughout ground tissue.

Fig. 1 (T.S of Curcuma longa) Fig. 2 (Large view of Cork cell)


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Powder Characteristics: The Powder of Curcuma longa rhizome is yellowish brown,
with aromatic and characteristic odour and slightly bitter in taste consist of cork, cortex,
fragments of thin walled parenchymatous cells, thick walled covering trichomes filled with
starch grains and oleoresins.

(Thin walled Parenchymatous Cell) (Thick walled covering trichome) (oleoresins)

(Stach grains) (Cork Cell) (Cortex)

Micrometry: The results of micrometric characters of tissue, cells and cell contents were
depicted in Table No.2. Measurements of different cells are frequently necessary for the
quantitative identification of closely allied substances. In most cases these allied substances
are mixed with the original drugs as adultrants and substituents. Thus, the adulterants and/or
substituent in crude drugs can be distinguished by this way with the aid of optical
microscopy. (fig.4)

Table 2: Micrometry of some cells

S.No Type of Cells Dimension Area

1. Cork Cell 5.36

2. Cortex 30.5

3. Oleoresin 3.25

4. Epidermal Cell 3.56

5. Fragments of groung 35.5


tissues

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Physicochemical Evaluation: The results of the physicochemical constants of raw material lie
within the limit which is mentioned in Table No.3. This signifies that the quality and purity of
raw material was good enough. The results of foreign organic matter denote presence of any
organism, part or product of an organism other than that named in the specification and
description. As the ash values of the crude drugs lies within the fair limit which signify its
quality and purity and gives idea about the total inorganic content. The water soluble
extractive value indicated the presence of sugar, acids and inorganic compounds and alcohol
soluble extractive values indicated the presence of polar constituents like phenols, alkaloids,
steroids, glycosides, flavonoids which signify the nature of the phytoconstituents present in
plant. As the pH also determined that is in acidic range and may be because of acidic salts
present in the rhizomes.Table 3: Physicochemical parameters

S.No. Parametrs (%w/w) Observations

1. Foreign Organic Matter 0.25

2. Moisture Content 9.52

3. Total Ash 8.35

4. Acid Insoluble Ash 0.95

5. Water Soluble Extractive value 25.15

6. Alcohol Soluble Extractive value 6.75

7. pH 03.00

Preliminary Phytochemical Screening: The Preliminary Phytochemical Investigations of


Aqueous extract, acetone extract, ethanolic extract and methanolic extract of Curcuma longa
rhizome were preformed which reveals the presence of Phenolic compound, Tannins,
Alkaloids, Terpenes, Saponin type of major secondary metabolites which revealed their
potent therapeutic activity. The results of the screening were expressed in Table no.4.

Table 4: Preliminary Phytochemical Screening

S.No Extract Constituents Ethanolic Extarct Aqueous Extract

1 Carbohydrates + ++

2 Proteins + +

3 Flavonoids + +

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4 Tannins + +

5 Steroids + +

6 Glycosides + +

7 Saponins + +

8 Alkaloids + +

9 Triterpenoids + ++

10 Amino acids - +

Florescence Analysis: The powder drug of the rhizome part of Curcuma longa (mesh size
40) was examined under daylight and UV light. The observation was recorded as under Table
5.

Table 5. Florescence analysis of Curcuma longa powdered drugs

S.No Treatment Day light UV 254nm UV 366nm

1. Powder as Such Green Light green Black

2. Powder + 1N NaOH in methanol Green Light green Black

3. Powder + 1N NaOH in water Light Green Dark green Black

4. Powder + 50% HCL Dark Green Light green Dark black

5. Powder + 50% Sulphuric acid Light Green Light green Black

6. Powder + 50% Nitric acid Green Light green Light black

7. Powder + Petroleum ether Light Green Green Black

8. Powder + Chloroform Dark Green Green Light black

9. Powder + Picric acid Green Light green Black

10. Powder + 5% Ferric chloride Dark Green Light green Light black
solution
11. Powder + 5% Iodine sol Light Green Green Black

12. Powder + Methanol Dark Green Green Black

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Market Sarvey for Standardized Ayurvedic fomulation of Curcuma longa

S.No Name of Constituent Present Name of Commonly


Formulation Manufactured Used

1. Kalyanavleha Curcuma longa, Cyclospermum CSIR National Commonly


ia an leptophyllum, Zingiber officinale, Research Lab, used for
Polyherbal Cuminum cyminum, Glycyrrhiza Lucknow. treating
formulation glabra, Acorus calamus, Piper Rheumatica
longum and Saussurea costus. and Arthiritic
disease.

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2. Curcumin 100 % standardized extract of pure Planet Ayurveda, Inflammatory
500 mg Curcumin. They contain US-FDA registered diseases
Capsules
95% of curcuminoid alkaloid for legal exports to
found packed in vegetable capsule USA
shells. OR
PLOT NO. 627,
JLPL,
INDUSTRIAL
AREA, SECTOR -
82, MOHALI -
140306, PUNJAB
(INDIA)

3. Boswellia Curcumin (500 mg) contains Planet Ayurveda, Ultimate


standardized extract of 250mg of US-FDA registered support for
Capsule
Boswellia serrata and standardized for legal exports to keeping
extract of 250 mg of 95% USA
healthy joints
curcuminoids obtained from OR
Curcuma longa. PLOT NO. 627, along with
JLPL, maintaining
INDUSTRIAL the overall
AREA, SECTOR - health of the
82, MOHALI - body.
140306, PUNJAB
(INDIA)

4. Harida Haridra contains a pale yellow to Himalaya The


orange-yellow volatile oil (6%) Pharmaceutical Pvt. curcuminoids
composed of a number of Ltd. Makali, contribute
monoterpenes and sesquiterpenes, Bengaluru, KA, towards the
including zingiberene. The 562 & 162, India antioxidant,
coloring principals (5%) are anti-
curcuminoids, 5060% of which inflammatory
are a mixture of curcumin, and
monodesmethoxycurcumin and cytoprotectiv
bisdesmethoxycurcumin. e properties
of Haridra.
5. Curcuma Standradized Curcuma Longa Herbal One Used for
longa Linn Linn Extract 500 mg. Company Ltd. relief of
100 Capsules- (Ouay-Un), atulence
Kamin Chun Bangkok, Thailand and
indigestion,
and as
treatment for
diseases of
the liver,
gallbladder,
pancreas and
stomach. It is
also natural
antibiotic and
antiseptic,
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with a very
effective
anti-
inflammatory
,
detoxificatio
n and
choleretic
action.
6. Turmerichills Haldi Extract (150mg) and Haldi Herbal Hills Plot Its anti-
Capsules Powder (150mg). No. 33 ABCD, inflammatory
Govt. Industrial property is
Estate Charkop, used in
Kandivali (W) treating
Mumbai common
cough &
cold. Its
strong
antimicrobial
property is
used as a first
line remedy
for cuts &
wound. It is
also known
as potent
lipolytic.
7. Vicco Turmeric and Sandalwood oil Viccoo The cream is
Turmeric Laboratories, 78, beneficial in
Cream- Farmland, fighting of
Turmeric Ramdaspeth, acne,
(Ayurvedic Nagpur 440 010, pimples,
Medicine Maharashtra, India. boils,
Cream) blemishes
and other
common
disorders of
skin.
DISCUSSION

As a part of standardization study, the macroscopical examination of CL was studied.


Macroscopical evaluation is a technique of qualitative evaluation based on the study of
morphological and sensory profiles of drugs. The macroscopical characters of the CL can
serve as diagnostic parameters. The extractive value, ash value, loss on drying and
fluorescent analysis of whole plant extracts have been carried out. The physical parameters,
such as loss on drying, ash values and extractive values will be helpful to identify the
authenticity of even from the crushed or powdered plant materials. It will serve as a standard
data for the quality control of the preparations containing this plant in future. The results

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showed greater extractive values in hot extraction, indicating the effect of elevated
temperature on extraction. Percentages of the extractive values were calculated with reference
to airdried drug. The percent extractives in different solvents indicate the quantity and nature
of constituents in the extracts. The extractive values are also helpful in estimation of specific
constituents soluble in particular solvent. It will serve as a standard data for the quality
control of the preparations containing this plant in future. The information obtained from the
ash values and extractive values are useful during the time of collection and also during
extraction process. Using these standards, the plant can be differentiated from other related
species. The plant may be considered as biosynthetic laboratory for a variety of compounds
(secondary metabolites) like alkaloids, glycosides, flavonoids, volatile oils, and saponins that
exert physiological effects. The fluorescence analysis of the powdered drug from the CL in
various solvents was performed under normal and UV light. Rhizome plant extracts are
examined in short UV (254nm) and long UV (366 nm) to detect the fluorescent compounds.
Phytochemical study was also useful to isolate the pharmacologically active principles
present in the drug. More phytochemical research work is required for isolation, purification
and characterization of biologically compounds.

CONCLUSION

Standardization is essential measure for quality, purity and sample identification.


Macromorphology and microscopy along with the Quantitative analytical microscopy is one
of the simplest and cheapest methods to start with for establishing the correct identity of the
source materials. Physicochemical and chemical analysis of rhizome confirm thequality and
purity of plant and its identification. The present study was useful for further pharmacological
and therapeutic evaluation along with the standadization of plant material.

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