Kalmeg (Andographis Paniculata)

SHAILENDRA SINGH RAJPUT
RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)
KALMEGH :1- INTRODUCTION :Andrographis paniculata, the Kalmegh of the Ayurveda, is an erect annual herb extremely bitter in taste in each and every part of the plant body. The plant is known in north-eastern India as Maha-tita, literally king of bitters, and known by various vernacular names . It is also known as Bhui-neem, since the plant, though much smaller in size has a similar appearance and has the same bitter taste as that of Neem (Azadirachta indica). In Malaysia, it is known as 'Hempedu Bumi' literally means 'bile of earth' since it is one of the most bitter plants that are used in traditional medicine. In Tamil it is called 'Sirunangai' or 'Siriyanangai'. The genus Andrographis consists of 28 species of small annual shrubs essentially distributed in tropical Asia. Only a few species are medicinal, of which Andrographis Paniculata is the most popular.
1.1- SYNONYM :Scientific Name(s): Andrographis paniculata (Burm.f.) Wall. Nees. Common Name(s): Kalmegh (Hindi), Kalmegha (Snaskrit), Chuanxinlian (Chinese), Kalupnath , Kirayat (Hindi), Mahatita (King of Bitters), Alui , Bhunimba , Bhui-neem , Yavatikta (Sanskrit), Sam biloto (Malay)
Botanical classification Kingdom : Plantae Division : Angiospermae Class : Dicotyledoneae Order : Tubiflorae Family : Acanthaceae Genus : Andrographis Species : paniculata Nees Vernacular names Assamese Arabic Marathi Bengali Oriya Chinese English Persian Gujarati Sanskrit Hindi Tamil Kannada Telugu Malayalam Indonesian Thai Lao Sinhalese Chiorta Quasabhuva Oli-kiryata Klmegh () Bhuinimba Chuan Xin Lian () Green chirayta, creat, king of bitters, andrographis, India echinacea Naine-havandi Kariyatu Klamegha (), Bhnimba () Kirayat Nilavembu, Sirunangai, Siriyanangai Nelaberu Nelavemaa( ) or Nelavepu meaning "Neem of the ground". "Nela" = ground and "vemaa" = neem. Nilavembu, Kiriyattu Sambiloto Fa-Talai-Jorn, Fah-talai-jone, Fah-talai-jon ( ) La-Sa-Bee Hn Kohomba / Heen Kohomba ( ), meaning "small neem," or Hn Bm Kohomba / Heen Bim Kohomba( ) meaning "small neem of the ground." Description :-
Andrographis paniculata grows erect to a height of 30110 cm in moist, shady places. The slender stem is dark green, squared furrows in with and cross-section longitudinal
wings along the angles. The lance-shaped leaves have hairless blades marking up to 8 centimeters long by 2.5 wide. The small flowers are borne in spreading racemes. The fruit is a capsule around 2 centimeters long and a few millimeters wide. It contains many yellow-brown seeds
1.2- DISTRIBUTION :A. paniculata is distributed in tropical Asian countries, often in isolated patches. It can be found in a variety of habitats, such as plains, hillsides, coastlines, and disturbed and cultivated areas such as roadsides, farms, and wastelands. Native populations of A. paniculata are spread throughout south India and Sri Lanka which perhaps represent the centre of origin and diversity of the species. The herb is an introduced species in northern parts of India, Java, Malaysia, Indonesia, the West Indies, and elsewhere in the Americas. The species also occurs in Hong Kong, Thailand, Brunei, Singapore, and other parts of Asia where it may or may not be native. The plant is cultivated in many areas, as well.Unlike other species of the genus, A. paniculata is of common occurrence in most places in India, including the plains and hilly areas up to 500 m, which accounts for its wide use. Since time immemorial, village and ethnic communities in India have been using this herb for treating a variety of ailments. 1.3- CULTIVATION :-
Cultivars (including improved Varieties) Comparative studies of plants from different localities have shown variation in the total alkaloids content. Local clones are generally cultivated as there is no named variety available.However, based on plant height, leaf length, leaf width, leaf stem ratio, leaf biomass ratio and dry biomass (herbage) yield /plant and chlorophyll content, morphogenetically diverse genotypes have been reported from Bhuvneshwar, Lucknow, Kakori and Assam. Method of Cultivation:
The period of this crop is 4 to 5 months. The sandy soil, which has sufficient percentage of microorganisms and has well ability to not accumulate the water, is better for its cultivation.
The 3 to 4 times ploughing of the land is sufficient before the sowing. Mix the natural Cow dung in field and make the lines. As it is the crop of rainy season therefore irrigate the crop, only when is required.
1.4- PREPARATION OF PLANTS IN NURSERY :In rainy season (around 15 June) for 1 acre, sow the 1 kg.Seeds in the one-meter wide lines by broad casting system and cover the seeds by thin layer of soil.Then irrigate the seeds.When plants become 1 month old (in July) then sow them in their permanent lines in field at the distance of 30 X 30 Cm (Line X Plant).It will be better if the sowing done during the dropping rain or irrigate the land with little volume of water after the sowing.There will be required 44,840 plants per acre (1,12,000 plants in one hectare).In rainy season there will grow many useless plants with the crop. Thus remove of the useless plants after the 30 and 60 days of sowing.It will increase the produce of Kalmegh Crop. Use 30 kg. Phosphorus,
30 kg. Potash and 60 kg. Nitrogen per hectare as fertilizer. Irrigate the crop essentially in the month of September if there is no any rain. By this the plants will increase properly.
Cutting and Drying: When the Kalmegh Plants is Green and has flowers then pull them out from the land with their roots. Remove the soil from roots and dry it on the dry and shaded place with little sunlight.Turn the plants after one week with awareness that leaves should not be off from the plant.When the plants are well dried put them in Jute bags and store. Besides Kalmegh we do have seeds of Tulasi (Ocimum Sanctom), Sanay ( Casia Angustifolia Vahl), Safed Musali (ChlorophytumBorivilianum), Isabgol (Plantago Ovata) Baibadag (Embelic Ribs Burm), Ashwagandha (Withania Somnifera) etc. Soil And Climate :It can be cultivated on wide range of soils from loam to lateritic soils with moderate fertility. It can be cultivated on shady wastelands also. The climatic requirement of the plant is hot and humid conditions with ample sunshine. With the onset of monsoon, plant grows luxuriantly and starts flowering with the moderation in temperature during September. Flowering and fruiting continues upto December until temperature drops drastically in Northern plains. Nursery Raising And Plantning Its propagation is through shattered seeds in nature. Vegetative propagation is also ossible in certain special cases through layering as each node is capable of roducing enough roots. Seeds are small and remain dormant for five to six months. For raising crop in one hectare three beds of 10x2 m size should be tilled, pulverized and levelled during the month of May. Liberal use of organic manure in nursery is advised for raising healthy seedling. Seeds should be
covered by very thin layer of soil and compost mixture. Beds should be covered properly by suitable mulch and irrigated regularly with water fountain till seedlings merge (6-7 days). Immediately after germination, mulch is removed to avoid elongation of the seedlings. After 10-15 days regular flood irrigation can be given till it becomes ready for planting. Transplanting of seedling is done in second fortnight of June at a row and plant spacing of 45 to 60 cm and 30 to 45 cm respectively. Beds should be irrigated immediately after planting. Thinning And Weeding To begin with one or two weeding/hoeing are essential to get the crop established. After establishment, crop grows well during monsoon and does not face any competition from weed. Manures, Fertilizers And Pesticides The medicinal plants have to be grown without chemical fertilizers and use of pesticides. Organic manures like, Farm Yard Manure (FYM), Vermi-Compost, Green Manure etc. may be used as per requirement of the species. To prevent diseases,nbio-pesticides could be prepared (either single or mixture) from Neem (kernel, seeds & leaves), Chitrakmool, Dhatura, Cow's urine etc. Irrigation Fairly distributed rainfall during monsoon is sufficient to raise annual crop in Northern states. But prior to rain 2-3 irrigations are required. Irrigation during autumn does not show much effect on biomass yield as by that time plant is in reproductive phase.
1.5- BOTONY :The herb of Andrographis paniculata is an erect, annual and sometimes it assumes height of shrub also. The flowers of the plant are insignificant and of dirty pink color [2].In Ayurveda the drug has been described as antipyretic and hepatoprotective. Cold infusion of the drug is mentioned in Sushruta Samhita for fever and liver disorders. Toxicity The possible testicular toxicity of Andrographis paniculata, standardized dried extract was evaluated in male Sprague Dawley rats for 60 days. No testicular toxicity was found with the treatment of 20, 200 and 1000 mg/kg during 60 days as evaluated by reproductive organ weight, testicular histology, ultrastructural analysis of Leydig cells and testosterone levels after 60 days of treatment. It is concluded that Andrographis paniculata dried extract did not produce subchronic testicular toxicity effect in male rats].
1.6- ACCORDING TO AYURVEDA IT CONTAIN :Rasa (Taste) Tikta (Bitter) Guna (Characteristics) Laghu (Light), Ruksha (Un-unctuous) Veerya (Potency) Ushna (Hot) Vipaka (Post digestion effect) Katu (Pungent) Actions according to Ayurveda: Deepana : Kalmegh ignites the digestive fire in stomach
Swedana: Kalmegh stimulates perspiration and removes toxins from the body via sweat. Krimighna: Kalmegh works well against parasites and microbes. Pittasaraka: Kalmegh stimulates expulsion of bile from liver. Yakrita Roge Prashasyate: Kalmegh is indicated in all disorders of liver. Kushthagyna: Kalmegh is useful in skin diseases 1.7- CHEMISTRY :Analysis of the whole plant gave the following lactones (dry basis): andrographolide,0.6%; 14-deoxy-11-oxoandrographolide (C20 H 28 O5), 0.12%;14deoxy -11, 12-ldehydroandrographohde (C20H30O4), 0.02% and a (C20,H30O4), 0.06%; non-bitter 14deoxyandrographolide constituent,
neoandrographolide (C26 H40 O8),0.005%. he leaves contain andrographolide (yield, 1%). From the petroleum ether extract of the leaves from Bangladesh, the following compounds have been isolated: a , b -unsaturated lactone, homoandrographolide (C22 H32 O3). andrographosterol (C23 H38 O), andrographane (C40 H82), andrographone (C32H640 ), a wax, and two esters containing, hydroxyl groups. he roots gave apigenin-7,4'-di-0-methyl ether, andrographolide and a new natural flavone, 5-hydroxy-7,8,2',3'-tetramethoxyflavone (C19H1807 );yield, 0.006%). They also contain a monohydroxy-trimethylflavone, andrographin (C18H1606 )and a dihydroxy-di-methoxyflavone, panicolin (C17H14O6 ). The presence of a - sitosterol is also reported.
Chemical markers Andrographolide, a diterpene lactone is the major constituents of the drug and its minimum quantity is prescribed as 0.5-0.9% in the appropriate drug.
Specification Andrographolide 5.0%
Chemical composition & pharmacology Three bitter principles, deoxyandrographolide, andrographolide and
neoandrographolide have been isolated from the whole plant and leaves1.The hepatoprotective effect of the water extract of the plant and the active principle
andrographolide has been studied 2. Both leaf extract and andrographolide in a single oral dose also inhibited the microsomal lipid peroxidation induced by carbon tetrachloride3. The effect of aqueous extract on the bile flow in albino rats was compared with phenobarbitone. Significant increase in liver weight was also observed4. 1.8- BIOLOGICAL ACTIVITY :

Carbohydrates Lipids Alkaloids Caffeic acid derivatives Flavonoids Iridoid glycosides Bitter glycosides Terpenoids Glucosinolates Vitamins
Other organic acids From the methanol extract the following organic acids were isolated: numeric acid, syringic acid, vanillic acid, p-hydroxy benzoic acid, ferulicacid, p-coumaric acid, gentisic acid, traces of salicylic acid, benzoic acid and cinnamic acid (Pailerand Haschke-Hofmeister, 1969).
1.9- MORPHOLOGY :An erect annual herb, stem dark green, 0.3-1.0 m in height, 2- 6 mm in dia., quadrangular with longitudinal furrows and wings on the angles of the younger parts, slightly enlarged at the nodes. Leaves glabrous, up to 8.0 cm Iong and 2.5 cm broad, lance late, pinnate; flowers small, in lax spreading axillary and terminal
racemes or panicles; capsules linear-oblong, acute at both ends, 1.9 cm x 0.3 cm long; seeds numerous, sub quadrate and yellowish brown in color.
Morphological Description:Andrographis is an herbal medicine derived from the Andrographis Paniculata shrub grown the moist, shady areas of India, China and throughout Southeast Asia. It is commonly referred to as 'Indian Echinacea' and is a popular Ayervedic and Chinese household remedy for the common cold, digestive issues, upper respiratory tract infections, flu and other sicknesses typified by fever. Although its use dates back easily a thousand years in these ancient medical traditions, there is a more recent historical reference to its potency. In the Indian flu epidemic of 1919, the herb Andrographis is credited with the reversal of its onslaught. More recently, it popularity has spread to Scandinavia, where its dosages have been standardized and it has been widely recommended by doctors for two decades as a common remedy in treating these same winter ailments.
Morphological characteristics of drugDrug consists of dried leaves and tender shoots; yields not less than 1% of andrographolide on dry wt. basis. Normally powdered drug is used. 1.10- MACROSCOPY :Commercial sample consists of chopped pieces of the whole plant with stem, roots, leaves, inflorescence stalk, flowers and fruits along with a few seeds. Stem Dark green, hard, quadrangular and narrowly winged in upper parts; glabrous about 6 mm thick; scars of detached leaves opposite and decussate; node swollen; fracture short; taste bitter.
Leaf
Present in broken pieces, fragile, deep green in ad axial surface, paler on the abaxial surface; winged, short petiole of about 8 mm long present; tip acute to acuminate. Flower Small, zygomorphic, pedicel late with a stalk of about a centimeter; solitary detached or intact on lax, spreading axillary or terminal racemes or panicles; bracts present, lance late, about 5 mm long; sepal linear lance late, glandular, pubescent; corolla two lipped with a slightly enlarged tube; generally white with purplish tinge; outer surface of petal hairy, upper lip two lobed, lower three lobed. Fruits A capsule; linear, pubescent with yellowish brown numerous seeds supported on retinacula; seeds sub quadrate rugose. 1.11- MICROSCOPIC :StemTS square in outline with four arrow winged; epidermis with a thin cuticle and few glandular trichomes; cortex narrow; collenchymatous tissue present in the wings and below the epidermis; rest of cortex is parenchymatous containing chlorophyll; epidermal cells contain cystoliths; endodermis present; phloem narrow with a few isolated
fibres and small accicular calcium oxalate crystals present occasionally; xylem wide, vessels scattered; accicular fibres and tracheids present; pith present small, with occasional cystoliths and acicular crystals of calcium oxalate. Leaf Petiole TS shows lateral wings; epidermis followed by Collenchyma and Chlorenchymatous tissue; two vascular bundle present in each of the wings on the ad axial side; meristele central in the form of an arc surrounded by parenchyma; small acicular crystals of calcium oxalate present in the parenchymatous tissue.
Midrib TS shows convex outline on the ad axial surface and a laterally flattened outline on the abaxial side, several cells of epidermis contains cystoliths, followed by collenchymas adjacent to both upper and lower epidermis, meristele similar to that of petiole. Lamina Dorsiventral; epidermal cells contains cystoliths; surface view of upper epidermis shows straight walled cells; with abundant diacytic stomata lower epidermal cells sinuous trichomes present, ordinary with one to three cells more on the margins; glandular with unicellular stalk and multicellular head present. Pallisade of one layer continues below the midrib also. PowderShows abundant simple, uni-and 2 to 3 celled multicellular short and long trichomes; glandular trichomes with short and long multicellular stalk and
multicellular head from leaf, stem and all parts of the flower; long acicular fibres from xylem region of stem, epidermal cells of the leaf and petiole in surface view embedded with diacytic stomata and cystolith; parenchymatous cells of the pith of stem embedded with acicular crystals of calcium oxalate and simple starch grains; fragments of spiral, pitted and scalariform vessels, polygonal thick-walled cells of the corolla tube, pollen grains and sclereidal layers from the stamen; fragments of cotyledon and endosperm, fibres and sclerosis from the fruit wall.
1.12- EXTRACTION :Air dried (3550C) leaves (1.0 g) of A. paniculata were extracted with solvents (3 _ 25 mol each) of varying polarity viz. chloroform, ethyl acetate, methanol and ethanol, in order to optimize the extraction method for the most efficient extraction of andrographolide and it derivatives. Extracts were concentrated under vacuum,redissolved in methanol, filtered, re-concentrated and finally made up to 5 mol volume with methanol prior toHPTLC analysis. Chromatographic condition6 Chromatography was performed on pre-activated (at 110C) silica gel 60 F254 HPTLC plates (10 _ 10 cm; 0.25 mm layer thickness).Samples and standard compounds were applied to the layer as 8 mm wide bands, positioned 10 mm from the bottom of the plate, using an automated TLC applicatorLinomat IV (Camag, Muttenx, Switzerland) with nitrogen flow providing delivery from the syringe at a speed of10s/mL. These critical parameters were maintained for all analyses performed. 1.13- IDENTIFICATION :-
Thin-layer chromatography: Carry out thin-layer chromatography on a precoated silica gel 60F254 plate (Appendix 3.5) using andrographolide and andrograpanin as reference
standards. Solvent system: Toluene : acetone : formic acid (6.0 : 4.0 : 0.4). Test solution: To 2 g of the substance being examined, add 25 ml of methanol, heat on a water bath for 10-15 minutes, cool and filter. Standard solution: Dissolve 10 mg each of andrographolide RS and 3 mg of andrograpanin RS in 10 ml of methanol. Procedure: Apply 10 l each of the test and standard solutions on a TLC plate as bands of 10 mm. Develop the plate to a distance of 8 cm from the line of application. Dry the plate in air and examine under 254 nm. Spray the plate with a solution of anisaldehyde sulphuric acid reagent. Heat the plate at 1100 for about 5 minutes till the bands are clearly visible. The chromatogram obtained with test solution shows a band at Rf ~0.60 corresponding to that of andrograpanin and at Rf ~0.40 corresponding to that of andrographolide and the profile should be similar to the one given in the TLC Experimental Dried leaves of A. paniculata were obtained from Malaysian Agricultural Research and Development Institute (MARDI) and were stored in a sealed air tight container at ambient conditions. Without any pre-treatment, the dried leaves were ground into powder and were sieved to obtain a particle size of less than 0.375mm. Carbon dioxide with a purity of 99.7% was used as the solvent in this study and was purchased in the liquid form from Air Product (M) Berhad. Analyses were done by high-performance liquid chromatography (HPLC) with a system from Shimadzu (Japan). Chromatographic peaks were identified by comparison with the retention time of the standards. Linear calibrations of standards at accuracy of more than 99% were carried out for the quantification of the A. paniculata extracts. Single injection of solvent (blank) was also done to determine the retention time of the solvent.
1.14- PURITY TESTS :General identity tests Macroscopic and microscopic examinations, chemical tests, and thin-layer chromatography for the presence of diterpene lactones (13). Microbiological Tests for specific microorganisms and microbial contamination limits are as described in the WHO guidelines on quality control methods for medicinal plants (12). Chemical Not less than 6% of total diterpene lactones, calculated as andrographolide(1, 3). Foreign organic matter Not more than 2% (1, 3). Acid-insoluble ash Not more than 2% (1, 3). Water-soluble extractive
Not less than 18% (1, 3). Alcohol-soluble extractive Not less than 13% using 85% ethanol (1, 3). Loss on drying Not more than 10% (1). Pesticide residues The recommended maximum limit of aldrin and dieldrin is not more than0.05 mg/kg (13). For other pesticides, see the European pharmacopoeia (13), and the WHO guidelines on quality control methods for medicinal plants (12) and pesticide residues (14). Heavy metals For maximum limits and analysis of heavy metals, consult the WHO guideline son quality control methods for medicinal plants (12). Radioactive residues Where applicable, consult the WHO guidelines on quality control methods for medicinal plants (12) for the analysis of radioactive isotopes. Other purity tests Total ash test to be established in accordance with national requirements. 1.15- STRENGTH :-
Physicochemical parameters
Foreign matter:
Not more than 2.0 Appendix 2.1.3
Loss on drying: Total ash: Acid-insoluble ash: Alcohol-soluble extractive: Water-soluble extractive:
per cent, Not more than 12.0 Appendix 2.1.4 per cent, Not more than 15.0 Appendix 2.1.5 per cent, Not more than 3.0 Appendix 2.1.7 per cent, Not less than 3.0 Appendix 2.1.8 per cent, Not less than 12.0 Appendix 2.1.9 per cent,
1.16- PHARMACOLOGY :Andrographolide isolated from Andrographis paniculata induces cell cycle Flavonoids and andrographolides from Andrographis paniculata. Two flavonoids, identified as 5,7, tetramethoxyflavanone and 5-hydroxy-7, trimethoxyflavone, as well as several other flavonoids, andrographolide diterpenoids, and polyphenols, were obtained from the phytochemical investigation of the whole plant of Andrographis paniculata, a well known medicinal plant.Anti-malarial activity of some xanthones isolated from the roots of Andrographis paniculata. Four xanthones were isolated from the roots of Andrographis paniculata. In vitro study revealed that compound 1,2-dihydroxy-6,8-dimethoxy-xanthone possessed substantial anti-plasmodial activity against Plasmodium falciparum Anticancer and immunostimulatory compounds from Andrographis paniculata. Hepatoprotective activity Andrographis paniculata (AP) treatment prevents BHC induced increase in the activities of enzymes y- Glut amyl transpeptidase , glutathione-S- transferase and lipid peroxidation . The activities of antioxidant enzymes like superoxide smutase , catalase , glutathione peroxidase and the levels of glutathione were decreased following BHC effect. Administration of AP showed protective effects in the
activity of superoxide dismutase , catalase , glutathione peroxidase , glutathione reductase as well the level of glutathione. The activity of lipid peroxidase was also decreased. The result indicates antioxidant and hepatoprotective action of A. paniculata. Alcoholic extract of the leaves of Andrographis paniculata was obtained by cold maceration. A dose of 300 mg/kg (1/6 of LD50) of the extract was selected to study hepatoprotective action against carbon tetrachlorideinduced liver damage. The extract was found to be effective in preventing liver damage which was evident by morphological, biochemical and functional parameters [5].Andrographolide from the herb Andrographis paniculata (whole plant) per se produces a significant dose (1.5-12 mg/kg) dependent choleretic effect (4.8-73%) as evidenced by increase in bile flow, bile salt, and bile acids in conscious rats and anaesthetized guinea pigs. The paracetamol induced decrease in volume and content of bile was prevented significantly by andrographolide pretreatment. It was found to be more potent than silymarin , a clinically used hepatoprotective agent. Hypoglycemic activity So far known Andrographis paniculata) is a hepatoprotective, ant platelet and antithrombotic drug. In this experiment its hypoglycemic effect has been tried in various ways. Water extract of AP 10 mg/kg body weight can prevent induction of hyperglycaemia significantly (P < 0.001) induced by oral administration of glucose 2 mg/kg body weight. But any how failed to do so in adrenaline induced hyperglycaemia . It also failed to demonstrate any "fasting blood sugar lowering effect" upon chronic administration (6 weeks) of AP. So probably AP prevents glucose absorption from gut. Whole experiment was done on rabbits. Anti -fertility activity
Anti-fertility effect of Andrographis Paniculata (AP) was observed in mice under
different experimental conditions. When sun-dried powder of AP was mixed with animal's food (Rats Pelletts ) in a dose of 2G per Kg. B.W. per day and was allowed to consume by the female mice daily for a period of six weeks, none of the animal (100%) were pregnant when mated with the male of proven fertility who did not receive the drug. On the other hand, the majority of the control female mice (95.2%) who did not receive the drug were pregnant when mated with the similar type of male as in the experimental group, and they continued to give birth to litters of usual size and number (average 5-6) for subsequent six matings [8].
Antivenin activity
Intraperitoneal injection of an ethanol extract of the aerial parts (25 g/kg bodyweight) to mice poisoned with cobra venom markedly delayed the occurrence of respiratory failure and death (6, 44). The same extract induced contractions in guinea-pig ileum at concentrations of 2 mg/ml. The contractions wereHerba Andrographidisenhanced by physostigmine and blocked by atropine, but were unchanged by antihistamines (44). These data suggest that extracts of the aerial parts do not modify the activity of the nicotinic receptors but produce significant muscarinic activity, which accounts for its antivenin effects (6, 44). Cardiovascular activity To observe the effects of Andrographis Paniculata component on nitric oxide, endothelin , cyclic guano sine monophosphate , lipid peroxide and superoxide dismutase in experimental therosclerotic rabbit model was established by feeding high cholesterol diet supplemented by bovine serum albumin injection bolus. The rabbits were randomly divided into the control, model, and andrographis paniculata treated group. Blood samples were collected before 4 weeks and 8 weeks after relevent treatment. Before 4 and after 8 weeks andrographis
paniculata administration, compared with model group, the nitric oxide, cyclic guano sine monophosphate and activity of superoxide dimutase increased, while lipid peroxide and endothelin decreased .it was concluded that andrographis paniculata possesses the effects of ant oxidation , preserving endothelial function, and maintaining the balance of nitric oxide/ endothelin [9].
Psycho- pharmacological activities Psychopharmacological studies were conducted on an extract of Andrographis paniculata herbs. The extract exhibited a significant alteration in behavior pattern and a reduction in spontaneous motility. The extract also produced a prolongation of the pentobarbitone -induced sleeping time and lowered the body temperature in different experimental animal models [10]. Anti -HIV activity A phase I dose-escalating clinical trial of andrographolide from Andrographis paniculata was conducted in 13 HIV positive patients and five HIV uninfected, healthy volunteers. The objectives were primarily to assess safety and tolerability and secondarily to assess effects on plasma virion HIV-1 RNA levels and CD4 (+) lymphocyte levels. No subjects used antiretroviral medications during the trial. Those with liver or renal abnormalities were excluded. The planned regimen was 5-mg/kg bodyweight for 3 weeks, escalating to 10-mg/kg bodyweight for 3 weeks and to 20-mg/kg bodyweight for a final 3 weeks. The trial was interrupted at 6 weeks due to adverse events including an anaphylactic reaction in one patient. All adverse events had resolved by the end of observation. A significant rise in the mean CD4 (+) lymphocyte level of HIV subjects occurred after administration of 10 mg/kg andrographolide (from a baseline of 405 cells/mm (3) to 501 cells/mm (3); p = 0.002). There were no statistically significant changes in mean
plasma HIV-1 RNA levels throughout the trial. Andrographolide may inhibit HIVinduced cell cycle dysregulation , leading to a rise in CD4 (+) lymphocyte levels in HIV-1 infected individuals [11]
Anti -allergic activity
One study, conducted by C?ceres , Hancke , et al., measured the effectiveness of Andrographis SHA-10 extract, compared against a placebo, in lessening the symptoms associated with the common cold. With a patient group of 158 male and female adults, the effects of Andrographis were measured on days 0, 2, and 4 of the treatment. On day 2, the patient group taking Andrographis displayed reductions in several of the associated symptoms, and on day 4, the same group was shown to have significant reductions in all of the symptoms as compared to the placebo group. In the summary conclusion, C?ceres , Hancke , et al. write, "It is concluded that Andrographis paniculata had a high degree of effectiveness in reducing the prevalence and intensity of the symptoms in uncomplicated common cold beginning at day two of treatment. No adverse effects were reported [12].In another study conducted by C?ceres , Hancke et al., one group of students in a rural school was given a placebo and another was given Kan Jang, a formulation of Andrographis paniculata from the Swedish Herbal Institute, and then observed to see how many colds occurred over a three month period. A dose of 200 mg a day was given to the study group, and after 1 month no significant difference was recorded. But after three onths, significant variances occurred. The Kan Jang group was 2.1 times less likely to catch a cold than the placebo group. They had a rate of incidence of 30 percent, compared to 62 percent in the placebo group. Anti Hepatic activity
The
aerial
parts
and
their
constituent
andrographolides
have
antihepatotoxicactivity in vitro and in vivo (4554). Intraperitoneal administration
of a methanol extract of the aerial parts (861.3 mg/kg body weight) to mice reduced hepatotoxicityinduced by carbon tetrachloride (CCl4), and reversed CCl4-inducedhistopathological CCl4-inducedincrease in the changes activity in of the liver (52). Intraperitoneal oxaloacetate administration ofandrographolide (100 mg/kg body weight) to mice inhibited the serum glutamate transaminase, serum glutamate pyruvate transaminase, alkaline phosphatase, bilirubin and hepatic triglycerides (52). Intraperitoneal administration of a methanol extract of the aerial parts (500 mg/kg body weight) to rats also suppressed the CCl4-inducedincrease in the activity of serum glutamate oxaloacetate transaminase, serum glutamate pyruvate transaminase, alkaline phosphatase and bilirubin (51). Intragastricadministration of an aqueous extract of the aerial parts (500mg/kg bodyweight) to ethanol-treated rats decreased the activity of serum transaminasesand suppressed histopathological changes in the liver (49). Andrographolide, the major antihepatotoxic component of the plant, exerted a pronounced protective effect in rats against hepatotoxicity induced by CCl4 (47), Dgalactosamine(54), was more paracetamol than (48) and ethanol the (49). Andrographolide effective silymarin, standard
hepatoprotective agent.
Ant malarial activity
50%
ethanol
extract
of
the
aerial
parts
inhibited
the
growth
of
Plasmodiumberghei both in vitro (100 mg/ml) and in mice after intragastric administration(1 g/kg body weight) (42). Intragastric administration of a 1-butanol, chloroform or ethanolwater extract of the aerial parts to Mastomys natalensis
inhibited the growth of P. berghei at doses of 12 g/kg body weight (43).Andrographolide(5 mg/kg body weight) and neoandrographolide (2.5mg/kg body weight) were also effective when administered by gastric lavage. Antibacterial activity
An ethanol extract of the leaves inhibited the growth in vitro of Escherichia coli and Staphylococcus aureus (28). A 50% methanol extract of the leaves inhibited growth in vitro of Proteus vulgaris (29). However, no in vitro antibacterial activity was observed when dried powder from the aerial parts was tested against E. coli, Staphylococcus aureus, Salmonella typhi or Shigella species. Anti-human immunodeficiency virus (HIV) activity
Aqueous extracts of the leaves inhibited HIV-1 infection and replication in the lymphoid cell line MOLT-4. A hot aqueous extract of the aerial parts reduced the percentage of HIV antigen-positive H9 cells. Dehydroandrographolideinhibited HIV-1 and HIV-1 (UCD123) infection of H9 cells at1.6mg/ml and 50mg/ml, respectively, and also inhibited HIV-1 infection of human lymphocytes at 50mg/ml. A methanol extract of the leaves suppressedsyncytia formation in cocultures of uninfected and HIV-1-infected MOLT cells (median effective dose [ED50] 70mg/ml). Immunostimulatory activity
Intragastric administration of an ethanol extract of the aerial parts (25mg/kgbody weight) or purified andrographolides (1 mg/kg body weight) to mice stimulated antibody production and the delayed-type hypersensitivity response to sheep red blood cells (35). The extract also stimulated a non-specific immune response in mice, measured by macrophage migrationindex,phagocytosis of[14C]leucine-
labelled E. coli, and proliferation of splenic lymphocytes (35). The extract was more effective than either andrographolide or neoandrographolidealone, suggesting that other constituents may be involved in the immunostimulantresponse (35). Antipyretic activity
Intragastric administration of an ethanol extract of the aerial parts (500mg/kgbody weight) to rats decreased yeast-induced pyrexia. The extract was reported to be as effective as 200 mg/kg body weight of aspirin, and no toxicity was observed at doses up to 600 mg/kg body weight. Intragastricadministration of andrographolide (100 mg/kg body weight) to mice decreased brewers yeast-induced pyrexia. Intragastric administration of deoxyandrographolide, andrographolide, neo andrographolide or 11,12-didehydro-14-deoxy andrographolide (100 mg/kg body weight) to mice, rats or rabbits reduced pyrexia induced by 2, 4-dinitrophenol or end toxins.
Antidiarrhoeal activity
Herba Andrographidis has antidiarrhoeal activity in situ. An ethanol, chloroform or 1-butanol extract of the aerial parts (300mg/ml) inhibited the WHO monographs on selected medicinal plants16E. coli enter toxin-induced secretary response which causes a diarrhoeal syndromein the rabbit and guinea-pig ileal loop assay. However, an aqueous extract of the aerial parts was not active. The constituent diterpenelactones, andrographolide and neoandrographolide, exhibited potentantisecretory activity in vivo against E. coli enter toxin-induced diarrhea. Andrographolide (1 mg per loop) was as active as lope amide when
tested against heat-labile E. coli enter toxin-induced diarrhea and more effectivethanloperamide when tested against heat-stable E. coli enter toxin induceddiarrhoea. Neoandrographolide (1 mg per loop) was as effective as lope amide when tested against heat-labile E. coli enter toxin-induced diarrhoea and slightly less active than lope amide when tested against heat-stable E. colienterotoxin-induced diarrhea. The mechanism of action involves inhibition of the intestinal secretory response induced by heat-abile E. coli enterotoxins,which are known to act through the stimulation of adenylate cyclase,and by inhibition of the secretion induced by heat-stable E. coli enterotoxins,which act through the activation of guanylate cyclase. Incubation ofmurine macrophages with andrographolide (150mmol/l) inhibited bacterialendotoxin-induced nitrite accumulation in a concentration- and timedependentmanner. Western blot analysis demonstrated that andrographolideinhibited the expression of an inducible is form of nitric oxide synthase linked to end toxin-induced circulatory shock .
Antiulcerogenic activity:-
P. major has been used in Turkey in the treatment of ulcers. The powdered dried leaves were taken together with honey daily before breakfast. A water immersionstress ulcer model was used on rats to test the plant extracts ability to inhibit ulcers. A test sample was given just before immobilization a stress cage. After 7 h immersed in water-bath the rats were killed and the stomachs were taken out for examination. The combined methanol- and water extract (1.2 g:kg) inhibited ulcer formation by 40% relative to the control group which received only the vehicle. The water extract (1 g:kg) inhibited ulcer formation by 37%and the
methanol extract inhibited it by 29%. P.major was not among the most active plants tested (Yesilada et al., 1993). Anticancer activity:In a screening of anticancer activity of Chilean plants a 50% ethanol extract of leaves, stems and seeds of P. major had no activity in vivo against lymphocytic leukaemia in mice (Bhakuni et al., 1976).A P. major preparation was reported to be effective in a screening system for prophylactic oncology. The effect included ant metastatic activity in models of tumour metastasis in mice. The details in this study were not described (Yaremenko,1990). In another study, an aqueous extract was shown to have a prophylactic effect on mammary cancer in mice (Lit hander, 1992). The leaves were extracted with phosphate buffer pH 7containing 0.9% NaCl and injected subcutaneously in mice of the C3H Strong strain. Among mice of this strain more than 90% develop cancer induced by a virus infection. After 60 weeks,93.3% of the untreated and 18.2% of the treated mice had tumours. The observed effect is thought to be due to stimulation of the immune system rather than a direct effect on the virus. No experimental results support this idea, only some observations made without experimental verification. The P. major extract had good effect on human herpes infections but had no effect on the herpes virus in in vitro tests. The same observations have been made for bacteria; only weak antibacterial activity of P. major extracts in vitro, but they hadan effect on infected wounds in vivo. While antibiotic son infected wounds had no effect, topical treatment with P. major extract eradicated the infections and healed the wounds. Immunomodulatory activity:The leaves extracted in saline for 2 h at 50C had chemo tactic activity on neutrophils using the Boyden migration chamber method, but it did not enhance neutrophil intracellular killing activity by the nitrozoblue tetrazolium reduction test.
1.17- USES :
Andrographis has been found to be an effective anti-biotic, anti-viral, antiparasitic and immune system stimulant.
It is used in viral hepatitis, children's bowel complaints, gastric acidity, liver congestion, flatulence.
It is being increasingly used as treatment for colds. Reasonably good evidence tells us that it can reduce the severity of cold symptoms.
It may also help prevent colds. Preliminary evidence suggests that it stimulate immunity, potentially making it useful for general immune support.
Interestingly, the ingredient of Andrographis used for standardization purposes, andrographolide, does not appear to affect the immune system as much as the whole plant extract.
Preliminary studies in animals also suggest that andrographis may offer benefits for preventing heart disease.
In addition, highly preliminary studies suggest that andrographis may help protect the liver from toxic injury, perhaps more successfully than the more famous liver-protective herb milk thistle. Decoction of the plant is blood purifier and is used for cure of torbid liver. Tincture of roots is tonic, stimultant and aperient.
Kalmegh has been used for liver complaints and fever, and as an antiinflammatory and immunostimulant. In clinical trials, Andrographis extract has been studied for use as an immunostimulant in upper respiratory tract infections and HIV infection. The potential of andrographolide as an anticancer agent is being investigated. However, clinical evidence to support the use of Kalmegh for any indication is lacking.
LITERATURE SURVEY :2- PHYTOCHEMICAL BASED : PAWAR R.K. et al (2011)10 A simple, rapid, selective and quantitative
HPTLC method has been developed for determination of Andrographolide in Andrographis paniculata (Whole Plant) and its formulationKalmegh Navaya Loha. The alcoholic extract of Andrographis paniculata (Whole Plant) and its ayurvedic formulationKalmegh Navayas Loha samples were applied on TLC Aluminium plate pre coated with Silica gel60 GF254 and developed using Toluene : Ethyl acetate : Formic acid (5:4.5:0.5) v/v
as
a mobile
phase. The
plate
was
sprayed
(derivatized)
with
Anisaldehyde Sulphuric Acid reagent followed by heating at 1100C for 10 minutes and detection and quantification were carried out densitometrically using an UV detector at wavelength of 235 nm. Content of marker compound Andrographolide found in the Andrographis paniculata (Whole Plant) and its formulation almegh Navaya Loha were 0.7746% w/w and 0.1155 % w/w respectively.
G. Ramesh et al (2011)3 Andrographis paniculata is a precious herb
belonging to the family Acanthaceae and possess valuable medicinal properties. A field experiment was taken up at Kannamangala Horticulture Farm, Kannamangala, Bangalore, Karnataka to study the interactive influence of organic and inorganic fertilizers on the growth and yield of kalmegh. The treatment of soil with 2.63 t ha-1 poultry manure + 150 % recommended dosage of fertilizers resulted in higher growth parameters in terms of plant height (42.0cm), number of leaves (107.8), number of branches ( 124.6), leaf area (102.0 cm2), plant spread (1131.4 cm2), fresh leaf weight (12.4 g), and higher yield parameters i.e., fresh weight per plant (35.6 g), dry weight per plant (13.4 g), fresh weight per plot (708.5 g) and herb yield (2.7 t ha-1). The treatments that received Azospirillum showed higher population of Azospirillum in soil and treatments that received VAM exhibited higher VAM root colonization when compared to the treatments that did not receive biofertilizers. The andrographolide content in plants subjected to VAM treatment was higher than the treatments that received either Azospirillum or organic fertilizers.
R. Radha et al (2011)2 The present study has been designed with the
objective to examine the petroleum ether, acetone, chloroform and methanol extracts of Andrographis paniculata leaves and stems, in order to evaluate the chemical composition, investigate its in vitro antimicrobial potential against strains of Enterococcus faecalis, Streptococcus
pyogenes, Klebsiella pneumonia, Proteus vulgaris, Candida albicans and Aspergillus flavus. Phytochemical analysis revealed the presence of flavonoids, alkaloids, glycosides, steroids, phenols, tannins and saponins. The antibacterial activity is more significant against Gram positive bacterium Enterococcus faecalis whereas the antifungal activity is more significant against Aspergillus flavus. These results may justify the popular use of this species as it has antimicrobial activity. However, in order to evaluate possible clinical application in therapy of infectious diseases, further clinical trials are required.
MAMATHA A.
et al (2011)6 A simple, sensitive and accurate HPTLC
method has been developed and validated for the quantitative estimation of Andrographolide in Kalmegh. The method employed TLC Aluminium plates precoated with silica gel 60 F254 as stationary phase with mobile phase as Chloroform: Methanol::7:1v/v. Andrographolide showed mean Rf value of 0.41 with max at 231nm. The method was validated in terms of linearity (100 500ng), precision, and accuracy (100.03% recovery). Limit of detection and limit of quantification were found to be 30ng and 100ng respectively. The validated method was applied for quantification of Andrographolide in methanolic extracts of Kalmegh samples obtained from various geographical sources. The proposed method is simple, sensitive and precise; it can be used for routine analysis.
Tanwer Babeet Singh et al (2010)11 Andrographis paniculata Nees
(Acanthaceae), commonly known as Kalmegh is traditionally used medicinal plant. In present study callus was raised from the seeds. Maximum callus was obtained on MS medium supplemented with 2, 4- D 4.52M/liter and IAA (5.71 M/l). The callus and different plant parts were used for primary metabolite quantification and antioxidant activity. Maximum soluble sugars and protein found in callus, maximum amount of starch, lipid and phenolic contents were found in leaf. Leaf extract showed
maximum superoxide radical scavenging activity (45.01.02%)) than other plant parts while root extract have maximum 2,2- diphenyl-1-picryhydrazyl (DPPH) free radical scavenging activity (71.421.14%)).
Ankita Kataky et al (2010)21 Since time immemorial the medicinal plants,
as sources of remedies, are widely used as alternative therapeutic tool for the treatment of plethora of acute and chronical diseases ranging from common cold to complex human diseases all over the world. Following the advent of modern medicine, herbal medicine suffered a set back, but due to the multiple drug resistance, side effects associated with antibiotics, restriction in use of synthetic antioxidant drugs because of its carcinogenicity and limited availability of anticancer drugs has forced the scientist to search for new alternative substances from plant origin. A. paniculata is in demand in terms of its high valued medicinal properties. Extensive research on this plant considered it as a good source of medicinal herb over the commercially available synthetic drugs. It can be used for pharmaceutical applications, food preservation and also as a food supplement, in order to promote growth of live stock and to increase the nutritional value of different foods and diet.
Ankita Kataky et al (2010)22 Antimicrobial activity of eight-months old
micro propagated plantlets of Andrographis paniculata was evaluated with various organic and aqueous extract against gram negative (Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa), gram positive (Staphylococcus aureus and Bacillus subtilis) bacteria and the fungal pathogen (Candida albicans). Agar well diffusion method was used to assess the antimicrobial activity of Andrographis paniculata. Chloroform extract showed strong inhibitory activity with all the microbes tested. Out of the five microbial test organisms Staphylococcus aureus was the most susceptible. The minimal inhibitory concentration (MIC) of the chloroform extract ranged from 15.625g/ml to 31.5g/ml. Phytochemical test reveals
the presence of carbohydrates, proteins, flavonoids, phenolics, saponin and alkaloids in the dried powder.
Thakur Gout am et al (2010)23 Diabetes mellitus (DM), a clinical
manifestation characterized by chronic hyperglycemia, is often ascribed to either a defect in insulin secretion, insulin resistance or both. Ayurveda (Indian Traditional Medicinal System) have shown promising results in the treatment of diabetes using various plants and herbs with negligible side effects and cost effective treatment. However, only limited number of these plants have been explored and scientifically validated for their hypoglycemic effect. This review highlights some of the plants being commonly used in India for their hypoglycemic effects.
MONIKA JADHAO
et al (2010)27 Herbal powder and poly herbal
formulation containing Andrographis peniculata were standardized and validated by high performance thin layer chromatographic method. Andrographolide in sample was identified and amount was estimated densitometrically. This method involves separation of compounds by TLC on pre-coated silica gel 60F 254 plates with the solvent system of benzene: ethyl acetate (5:5) and scanned using densitometric scanner in UV reflectance photo mode at 220 nm. The linearity was observed in the range of 360ng-660 ng /spot. The andrographolide content of 237.2 -g/ 100 mg in powder and 41.80 mg/ 5ml. was observed in the test sample. The average percentage recovery value of 97.68 was obtained. The proposed method being precise and sensitive can be used for the detection, monitoring and quantification of andrographolide in Andrographis peniculata.
Sunday J Ameh et al 201017 To characterize the aerial parts of
Andrographis paniculata, a bitter Indian herb grown in Nigeria, for the purpose of quality control. Methods: The determination of bitterness value
and
of
various
physicochemical
characteristics;
tests
for
key
phytochemicals; and thin layer chromatography (TLC) of the air-dried herb, were carried out as prescribed in standard texts. Results: The mean bitterness value of the herb for both men and women was 2.86 1.74 x 103 units per g. The male value (2.07 1.42 x 103) appeared to be lower than the females (3.52 1.82 x 103) but the difference was not statistically significant. The results (% w/w) of loss on drying (10.64 0.36), total ash (14.10 4.49), water extractive value (30.37 2.63) and acid insoluble ash (1.00 0.06) were similar to those reported for the Asian plant. The phytochemical tests revealed the presence of glycosides, saponins, tannins and alkaloids, but not of anthraquinones. Normal phase TLC of the drug yielded 5 spots as against 6 spots yielded by reverse TLC. Conclusion: The results provide useful quantitative and descriptive data that are essential for identifying and characterizing the Nigerian grown herb for the purpose of quality control; and confirm key similarities between the Nigerian and the Asian plant.
Andri Cahyo Kumoro
et al (2009)33
A solid-liquid extraction was
performed for the leaves of Andrographis paniculata in order to obtain two bioactive components, andrographolide and deoxyandrographolide. The problem of hydrolysble components and solvent removal difficulties in the conventional extraction led us to study further the effects of solvent properties on the conventional extraction using Soxhlet method in order to determine the best solvent or solvent mixture for high extraction yield of A. paniculata. This solvent selection is very important prior to alternative extraction methods since it can be used as a preextraction solvent, main solvent, or co-solvent. Based on the yield of extract and andrographolide and deoxyandrographolide content, methanol was found to be the best solvent
Andri Cahyo Kumoro et al (2009) A solid-liquid extraction was performed
for the leaves of Andrographis paniculata in order to obtain two bioactive components, andrographolide and deoxyandrographolide. The problem of hydrolysable components and solvent removal difficulties in the conventional extraction led us to study further the effects of solvent properties on the conventional extraction using Soxhlet method in order to determine the best solvent or solvent mixture for high extraction yield of A. paniculata. This solvent selection is very important prior to alternative extraction methods since it can be used as a preextraction solvent, main solvent, or co-solvent. Based on the yield of extract and andrographolide and deoxyandrographolide content, methanol was found to be the best solvent.
Praveen et al (2009)44 Adventitious roots were induced directly from leaf
segments of Andrographis paniculata on Murashige and
Skoog (MS)
medium with 5.3 M -naphthalene acetic acid (NAA) and 30 g/l sucrose. Adventitious roots cultured in flasks using MS liquid medium with 2.7 M NAA and 30 g/l sucrose showed higher accumulation of biomass (fresh and dry weight) and andrographolide within four weeks. Seven-fold increment of fresh biomass was evident in suspension cultures along with 3.5-fold higher andrographolide compared to natural plants. These results showed a great potentiality of adventitious root cultures for the production of andrographolide.
Phani Ratna Prasanth .G et al (2009) Around 70% of population in India
relies on these systems for primary health care. Ayurveda and Indian System of Medicine (ISM) utilize a vast number of plants. The traditional Indian system of medicine includes many natural plants used for the treatment of diabetic activity. In the present study, the herbal remedies used for diabetic treatment in Krishna district, which is located in coastal region of Andhra Pradesh, India are discussed. A total number of around
twenty-five plants such as Eugenia jambolana, Ocimum sanctum, Phyllanthus amarus, Pterocarpus marsupium, Tinospora cord folia, Curcuma longa and Trigonella forename graecum etc. from various families used therapeutically against diabetes and related disorders are being covered, which includes Biological source, family, parts of the plant used, regional name, local use and therapeutic action (traditional use) for all the natural plants. This detailed information will be helpful for the pharmacognosist, botanist, phytochemist and pharmacologist for the collection and identification of the plant for their research work.
K. Vijaya Lakshmi et al (2009) CPAC 5-7 and 7-7 of T1 generation and
plants CPAC 20-7-7 of T2 generation showed significant reduction in larval weight gain compared with non-transformed plants. Some of the plants of T1 and T2 generation of cry1Ac showed resistance and moderately resistance. In the present study, some of the plants of the progeny of T0 CPAC 1, 5, 7, 8, 9, 19 and 20 affected the larval weight gain, but consistency was not observed in the antibiosis performance against H. armigera larvae in subsequent generations. It is presumed that, the physiology of the plant, interaction of toxic protein with acid metabolic cycle with in the plant and internal gut environment of the larva due to consumption of the acid exudates of the plant may influence the potency of the Bt toxin in the chickpea. Hence, to produce transgenics with higher level of expression of Bt toxins in chickpea plants, the research need to be oriented with due Consideration to all the above factors
Shiv Narayan Sharma
et al (2009)30 In the present investigation, 15
Andrographis paniculata genotypes collected from Chhattisgarh and adjoining states were grown in the field for the variability studies. Three different variability parameters namely morphological, molecular and chemo typing of active ingredient content were employed. Wide variations were recorded with regard to quantitative characters and RAPD profile.
Ten polymorphic RAPD primers produced a total of 37 amplicons, which generated 70.27% polymorphism. The number of amplified products ranged from 2 to 7 for different primers, whereas the percentage of genetic similarity for the studied primers ranged from 51.4 to 97.0. There were two major clusters formed in the genotypes studied. The andrographolide content ranged from 0.69 to 1.85% and the genotype KI-2 had the highest estimated content. The study demonstrated that simultaneously morphological, molecular and biochemical analysis are useful for characterizing genetic diversity and defining relationships between kalmegh germplasm. It also gave possible indications to the phytochemical variation of different genotypes which were due to the genetic differences.
2.1- PHARMACOLOGY BASED : Lather A et al (2011)7 A large proportion of the population suffers from
piles. The term piles refers to thehaemorrhoidal swelling. Treatment of piles depends on the degree of their occurrence and their related symptoms. Conservative treatment with diet and drugs is of reventive and to a lesser degree of curative value. Local application of various ointments, sclerosal injection therapy, rubber band legation and cryosurgery can be used with advantage. Many indigenous drugs have been used in India for many decades in the treatment of piles. The home treatment of piles and the herbal treatment of piles are the alternative of the present surgical methods. This article is concerned with the use of herbs in the treatment of piles along with common name of the plant, their family, part to be used in the treatment of the piles.
Matthew I. Arhewoh et al (2011)5
The powdered leaves of Anona
muricata (AME), and Andrographis paniculata (APE) and seeds of Garcinia kola (GK) were separately subjected to ultrasonic and conventional (maceration) extraction methods. The antioxidant potency of their extracts were evaluated on the basis of their DPPH (2,2-Diphenyl-1picrylhydrazyl) scavenging activity. The results showed that the extraction yield using ultrasound-assisted method was 39.8, 34.3 and 21.2% for APE, GK and AME, respectively. The corresponding values for maceration method were lower: 15.6, 18.7 and 1.5%, respectively. The antioxidant (DPPH scavenging) activity of the extract obtained by ultrasound-assisted extraction was higher than that obtained by maceration. The
phytochemical components of the extracts include phenolics and flavonoids which might have contributed largely to the free radicals scavenging activity of the extracts. The findings of this work lend support to the claimed efficiency of ultrasonic extraction as well as to the use of these plants as health-promoting agents.
Shahid Akbar et al (2011)8 Andrographis paniculata is a plant that has
been effectively used in traditional Asian medicines for centuries. Its perceived blood purifying property results in its use in diseases where blood abnormalities are considered causes of disease, such as skin eruptions, boils, scabies, and chronic undetermined fevers. "e aerial part of the plant, used medicinally, contains a large number of chemical constituents, mainly lactones, diterpenoids, diterpene glycosides, flavonoids, and flavonoid glycosides. Controlled clinical trials report its safe and effective use for reducing symptoms of uncomplicated upper respiratory tract infections. Since many of the disease conditions commonly treated with A. paniculata in traditional medical systems are considered self-limiting, its purported benefits need critical evaluation. "is review summarizes current scientific findings and suggests further research to verify the therapeutic efficacy of A. paniculata
MAMATHA A et al (2011)6 A simple, sensitive and accurate HPTLC
method has been developed and validated for the quantitative estimation of Andrographolide in Kalmegh. The method employed TLC Aluminium plates precoated with silica gel 60 F254 as stationary phase with mobile phase as Chloroform: Methanol::7:1v/v. Andrographolide showed mean Rf value of 0.41 with max at 231nm. The method was validated in terms of linearity (100 500ng), precision, and accuracy (100.03% recovery). Limit of detection and limit of quantification were found to be 30ng and 100ng respectively. The validated method was applied for quantification of Andrographolide in methanolic extracts of Kalmegh samples obtained from
various geographical sources. The proposed method is simple, sensitive and precise; it can be used for routine analysis.
R khabiya et al (2010)
Medicinal herbs are significant source of
pharmaceutical drugs. Latest trends have shown increasing demand of phytodrugs and some medicinal herbs have proven hepatotprotective potential. Silymarin, a flavones lignan mixture) extracted from the Silybum marianum (milk thistle) is a popular remedy for hepatic diseases. Today every herbal company is marketing formulations for liver disorders but the actual scene is that only selected medicinal herbs have been tested for hepatotprotective activity. Some herbal formulations claiming to be hepatoprotective may actually contain chemical constituents having hepatotoxic potential. Andrographolide (Andrographis paniculata), Glycyrrhizin (Glychyrrhiza glabra), Picrrorihzin (Picrorrhiza kurroa) and Hypo-phyllanthin (Phyllanthus niruri) are potential candidates with hepatoprotective activity. The article reviews latest trends in testing of isolated constituents with hepatoprotective activity.
N. Abu backer et al (2010) The present study mainly focuses on the
potential antibacterial value of ethanol leaf extract of Andrographis paniculata against pathogenic bacteria Escherichia coli, Klebsiella peneumoniae, Proteus vulgaris and Streptococcus pneumoniae by disc diffusion method. Bioactive compound andrographolide was isolated from the leaf. The results revealed that the ethanol leaf extract and andrographolide compound are potent in inhibiting these bacteria and this work highlights that the inhibitory effect is on par with standard antibiotics.
Shewta Mishra et al (2010)19
Andrographis paniculata (Burm.f.) Nees
(Acanthaceae) is a medicinal plant traditionally used for the treatment of cold, fever, laryngitis and several infectious diseases ranging from malaria to dysentery and diarrhoea. The plant is widely used in Ayurvedic and
Homeopathic systems of medicines. The medicinal value of this plant is due to the presence of active ingredients viz andrographolide and neoandrographolide which are derivatives of diterpenoids. The content of these active ingredients in plant varies with in plant parts and with the geographical distribution. In order to study the variation in andrographolide content, plant material was collected from 15 districts of Madhya Pradesh and evaluated through a simple, quick and accurate HPLC method using C-18 ODS-2 column. The chemo profiling study showed significant variations in the concentration of active ingredients in the leaves as well as in whole plant. The study also revealed that andrographolide content was maximum in leaves of the accession collected from Seoni(1.82%) followed by Chhindwara (1.48%).
C. K. K. Nair et al (2010)16 According to one of the ancient proverbs in
India, there is no plant on earth which has no medicinal property. More than 90% of the compounds used in modern medicine have their origin from plant sources. A large number of plants have been used by man from ancient times as medicine for curing various ailments. In recent times there is an upsurge of interest and focus on the importance of medicinal plants and traditional health systems in solving the health care problems of the world. This article gives an over view and a brief account of the important ethno-medicinal plants used in treating neoplastic diseases.
KAPADI C. et al (2010)25 The present paper deals with development and
standardization of HPTLC method used for quantification of 14Deoxy 11,12didehydroandrographolide in Andrographis paniculata commonly known as Kalmegh, a plant of the family Acanthaceae. It is an Andrographolide derivative, isolated from the whole plant of Andrographis paniculata, which possesses immune stimulatory activity. Present work focuses on the development and standardization of a chromatographic method for quantification of 14Deoxy11,12didehydroandrographolide,
for the standardization of A. paniculata by HPTLC method. A simple, precise, accurate and rapid high performance thin layer method has been developed an dvalidated forth estimation of 14Deoxy11, 12 didehydroandrographolide. The lowest detectable limit of 4deoxy11, 12 didehydroandrographolide was found up to 40 ng with a good resolution and separation of the marker from the other constituents of A. paniculata. Further, the recovery value of 14deoxy11,12didehydroandrographolide was found to be about 94.02 96.34 %, which shows the reliability and suitability of the method. The structure of isolated marker was characterized and confirmed by various advanced spectroscopic methods
Pritam S. Jain
et al (2010) The present study was carried out to
determine the, anti-inflammatory activity of petroleum ether and methanol extracts of Wrightiatinctoria woody stems using animal models. The airdried, powdered woody stems (900 g) were extracted in Soxhlet apparatus with petroleum ether and methanol. The crude dried petroleum ether extract (12 g) and methanol (18 g) were prepared containing doses of 100, 200 and 400 mg/kg, and subjected to anti-inflammatory (carrageen anand histamine-induced paw edema test) assays. The results obtained indicate that the extract possessed significant dose dependent antiinflammatory activity. This study showed that the petroleum ether and methanol extracts of Wrightia tinctoria woody stems possess potential pharmacologically active constituents capable of inhibiting inflammation.
S.
J. Ameh et al (2010)14 CONAVIR is an immunostimulant
phytomedicine developed at the National Institute for Pharmaceutical Research and Development, Abuja. The paper describes the steps taken and the results obtained in the attempt to develop specifications for the herbal component the aerial parts of Andrographis paniculata Ness (Family: Acanthaceae), grown in the Institutes gardens. The said steps
are vital for production as per good manufacturing practice. Qualitative and quantitative tests on the fresh and air dried materials were carried out as prescribed in standard texts. Quantitative results on the dried herb included: bitterness value, 2.86 1.74 x 103 units per g; loss on drying, 10.64 0.36 %w/w; total ash, 14.10 4.49 %w/w; acid insoluble ash, 1.00 0.06 %w/w; and water extractable matter, 30.37 2.63 %w/w. Tests for phytochemicals revealed the presence of glycosides, saponins, tannins and alkaloids, but not of anthraquinones. Normal phase thin layer chromatography of the aqueous or ethanolic extract gave 5 spots, while the reverse phase of the same extracts gave 6. These results, including macroscopic and sensory examinations of the fresh herb, provided the quantitative and descriptive data needed for identifying and characterizing the herbal component of CONAVIR.
Reinhard Bhm et al (2009) The increased consumer demand for high
quality,
long
shelf-life,
and
ready-to-eat
foods
has
initiated
the
development of only mildly preserved products that keep their natural and fresh appearance as far as possible. Microbiological growth commonly induces undesirable organoleptic and appearance change during the storage of food products. If bacterial growth in food products could be delayed, or halted, large gains in products shelf-life would be possible.
Chauhan J.S. et al (2009)38 Andrographis paniculata (Burm. f.) Wallich ex
Nees contains a bitter compound called andrographolide is commonly used for medicinal purposes. The herb has been revered for treating infectious diseases and highly regarded also as having a preventative effect from many diseases, due to its powerful immune strengthening benefits. The propagation of A. paniculata through seed is somewhat difficult which may be due to various germination problems. In view of the above, the present investigation was conducted with an objective to
assess the compatible substratum for standard seed germination test of Andrographis seed. The seeds were collected and stored for six months at optimum temperature. Four substrata namely filter paper, top soil, saw dust and riverbed sand was collected for the experiment. Sand and soil were sieved with a 2 mm sieve to ensure uniform particle size. A standard germination test was done under partly sterilized and non-sterilized substratum with proper maintenance of light (11 hrs) and temperature (250C) by adding water up to 60 70 % WHC. The 16 days test showed varied germination index and number of secondary roots. The result showed that the sterilized filter paper has proved most effective and compatible substratum for the Andrographis paniculata standard seed germination test. This led to uniform and quick germination with proper secondary roots and primary leaves. It can be suggested that due to scarcity of filter papers, sand substratum can also be used as an alternate as it showed more uniform germination than the rest two substrata. [The Journal of American Science. 2009; 5(6):70-75]. (ISSN 1545-1003).
Pratibha Singh et al (2009) Kidney Disease is an increasingly common
condition with limited treatment options placing a major financial and emotional burden on the community. There is emerging evidence in the literature about renoprotective complementary and alternative medicines. A primary goal of this article is to present the scientific evidence for the use of herbs like Andrographis paniculata (AP) as a complementary treatment for acute renal failure (ARF). Wistar rats were divided as follows: Normal, Gentamicin (GM) and Root extracts (Pt. ether, CHCl3 and MeOH) of AP (200 mg/kg; p.o.) treated groups. The nephrotoxic model was prepared by GM (80 mg/kg; i.p. 8 days). The degree of nephroprotection was measured by using renal parameters like serum creatinine (SCr), serum urea (SU) and urinary proteins (UP) after 3, 7 and 10 days of ingestion of various extracts. Oral administration of Pt. ether (p<0.05), CHCl3 (p<0.01) and MeOH (p<0.001) extracts of AP patently prevented
gentamicin induced elevated levels of SCr, SU and UP. The results were also supported by measuring the urine volume voided by each rat separately, of all the groups with time. The extent of protection offered by various extracts under study increased with the increasing time of treatment and polarity of the solvents. The signs of GM nephrotoxicity in rats are significantly mitigated by Pt. ether and CHCl3 extracts whereas the maximal alleviation of ARF was caused by MeOH root extract; hence, the methanolic root extract of AP can be advocated as a nephroprotective agent.
2.2- MIXED LITRAETURE : S.Savitha et al (2011)1 This work mainly focus on the potential
antibacterial value of various extracts of Andrographis paniculata against various pathogenic microorganisms (E.coli, Klebsilla and Pseudomonas) by employing minimal inhibitory concentration. In siddha system of medicine the herb Nilavembu (A. paniculata) used to treat various ailments including infection diseases even viral origin too according to its phamacodynamics. Minimal inhibitory concentration values were compared with control the result revealed that the methanol extract is potent in inhibiting bacterial growth of gram negative organism. Medicinal plants are a source for a wide variety of natural antioxidants. Free radical scavenging potential of various extract (methanol, ethanol, and aqueous) of the Andrographis paniculata, a medicinal plant was evaluated by using reducing power. The reducing power method ascorbic acid was used as a
standard for determining reducing power. Significant results were observed in the methanol extract has estimated parameters. The methanol extracts of the plant exhibited appreciable activity as compared to the aqueous and ethanol extract. There by indicating that the Andrographis paniculata has promising free radical scavenging activity.
R. Radha et al (2011)2 The present study has been designed with the
objective to examine the petroleum ether, acetone, chloroform and methanol extracts of Andrographis paniculata leaves and stems, in order to evaluate the chemical composition, investigate its in vitro antimicrobial potential against strains of Enterococcus faecalis, Streptococcus pyogenes, Klebsiella pneumonia, Proteus vulgaris, Candida albicans and Aspergillus flavus. Phytochemical analysis revealed the presence of flavonoids, alkaloids, glycosides, steroids, phenols, tannins and saponins. The antibacterial activity is more significant against Gram positive bacterium Enterococcus faecalis whereas the antifungal activity is more significant against Aspergillus flavus. These results may justify the popular use of this species as it has antimicrobial activity. However, in order to evaluate possible clinical application in therapy of infectious diseases, further clinical trials are required.
G. Ramesh et al (2011)3 Andrographis paniculata is a precious herb
belonging to the family Acanthaceae and possess valuable medicinal properties. A field experiment was taken up at Kannamangala Horticulture Farm, Kannamangala, Bangalore, Karnataka to study the interactive influence of organic and inorganic fertilizers on the growth and yield of kalmegh. The treatment of soil with 2.63 t ha-1 poultry manure + 150 % recommended dosage of fertilizers resulted in higher growth parameters in terms of plant height (42.0cm), number of leaves (107.8), number of branches ( 124.6), leaf area (102.0 cm2), plant spread (1131.4 cm2), fresh
leaf weight (12.4 g), and higher yield parameters i.e., fresh weight per plant (35.6 g), dry weight per plant (13.4 g), fresh weight per plot (708.5 g) and herb yield (2.7 t ha-1). The treatments that received Azospirillum showed higher population of Azospirillum in soil and treatments that received VAM exhibited higher VAM root colonization when compared to the treatments that did not receive biofertilizers. The andrographolide content in plants subjected to VAM treatment was higher than the treatments that received either Azospirillum or organic fertilizers.
Somnath Ghosal et al (2011) Non-Timber Forest Products (NTFPs) have
an important role in the household economy of forest fringe dwellers living in the dry-deciduous forests of Purulia, Bankura and West Midnapur districts of West Bengal, India. Due to the lack of agricultural land and industrial activities, forest fringe people collect forest products in regular basis for their livelihoods. They also make some value-added products to have some extra money. It is comparatively easier to calculate the monetary value of NTFPs, which are marketed at the local market or by intermediaries. However, the products which are normally collected for domestic uses carry enough monetary value too, but it never calculated in a proper way. It is very difficult to get a proper information from forest villagers that which product how much each household collect annually as the production of NTFPs vary from one year to another. A relative value can be calculated based on the importance, availability and quantity of these products.
Renu Parashar
et al (2011)9 A field experiment was conducted on
Andrographis paniculata (during Kharif season 2007), atDusty acre farm of JNKVV, Jabalpur. The experiment was laid out using randomized complete block design with four replicates. The samples were collected at seven different growth stages. The results were evaluated on the basis of growth analytical parameter (viz, Leaf area, Leaf area index, Crop growth
rate, Relative crop growth rate, Leaf area duration, Biomass duration and Specific leaf area) as well as morpho-physiological (viz, Plant height, No. of leaves, No. of branches, plant fresh weight, plant dry weight, leaves fresh and dry weights and herbage yield) parameters. The result revealed that all the selected parameters goes on increasing with crop growth stages. However, the specific leaf area increases up to 90 DAS and there after it gradually decline up to 120 DAS indicating the maturity of crop
Bhasker Joshi et al (2011) The medicinal properties of forest vegetation
was analyzed in a submontane forest of Tarai and Bhawar of Kumaun adjacent to Kashipur, at (29 14-43.6)(29 19-50.5) E longitude and (79 03-22.6)(79 04-23.2) N latitude at an elevation of 253.4265.5 meter above the sea level, within the districts of Nainital and Udham Singh Nagar. 29 plants species belonging to 22 family, 26 genera, and 29 species were reported. Of these leaves in 19% cases, roots and whole plants in 16% cases, fruits and bark 13% cases are used. Based on life form 17 phanerophytes, 5 chamaephytes, 4 therophytes, 2 hemi cryptophytes and 1 therophyte were recorded.
P.D. MAKWANA et al (2010) Kalmegh (deshi kariatu) is a substitute of
Nepali kariatu (swita chiruta). Kalmegh can easily grown in the Gujarat as wild. Due to short supply and poor quality of kalmegh, it is necessecery to cultivate in the field. Regular cultivation of kalmegh is started in the Gujarat during the year 2002. Ayurveda is an ancient science of life. Which has a strong philosophical basis. Ayurveda is a dynamic phenomenon that offers multifaceted approaches for healing. It comprises of knowledge about the plants that are primarily based on the past experiences and present uses in India, more as living tradition. Significantly higher andrographolide content was observed under the treatment of organic manures i.e. FYM 10 t/ha and Spacing i.e.30 x 45 cm (1.39 and 1.40%, respectively). Interaction effect of FYM @ 10 t/ha with wider spacing of 30cm x 45cm had recorded
higher
1.5%
andrographolide
content
noted
higher
amount
of
andrographolide content. Application of FYM 10 t/ha also recorded significantly higher organic carbon , available N, P and K.
Shiv Narayan Sharma et al (2010)
Botanical pesticides are good
alternatives to chemical pesticides. It is eco-friendly, economic, targetspecific and biodegradable. Present day research necessitate to explore and harnessing the traditional knowledge regarding the use of natural products for crop protection in reference to traditional folk practices prevalent among farmers as well as information from classical literature. This review provides a brief explanation of various green pesticide research methodologies that are helpful in and quick identification, upgradation, standardization of botanicals.
ADI C.
at el (2010)
The present paper deals with development and
standardization of HPTLC method used for quantification of 14Deoxy 11,12didehydroandrographolide in Andrographis paniculata commonly known as Kalmegh, a plant of the family Acanthaceae. It is an Andrographolidederivative, isolated from the whole plant of Andrographis paniculata, which possesses immunostimulatory activity. Present work focuses on the development and standardization of a chromatographic method for quantification of 14Deoxy11,12didehydroandrographolide, for the standardization of A. paniculata by HPTLC method. A simple, precise, accurate and rapid high performance thin layer chromatographic method has-been developed and validated for the estimation of 14Deoxy 11,12didehydroandrographolide. The lowest detectable limit of 14deoxy 11,12didehydroandrographolide was found up to 40 ng with a good resolution and separation of the marker from the other constituents of A. paniculata. Further, the recovery value of 14deoxy11,12 didehydroandrographolide was found to be about 94.02 96.34 %, which
shows the reliability and suitability of the method. The structure of isolated marker was characterized and confirmed by various advanced spectroscopic methods
Sunday J Ameh
et al (2010) To characterize the aerial parts of
Andrographis paniculata, a bitter Indian herb grown in Nigeria, for the purpose of quality control. The determination of bitterness value and of various physicochemical characteristics; tests for key phytochemicals; and thin layer chromatography (TLC) of the air-dried herb, were carried out as prescribed in standard texts. The mean bitterness value of the herb for both men and women was 2.86 1.74 x 103 units per g. The male value (2.07 1.42 x 103) appeared to be lower than the females (3.52 1.82 x 103) but the difference was not statistically significant. The results (% w/w) of loss on drying (10.64 0.36), total ash (14.10 4.49), water extractive value (30.37 2.63) and acid insoluble ash (1.00 0.06) were similar to those reported for the Asian plant. The phytochemical tests revealed the presence of glycosides, saponins, tannins and alkaloids, but not of anthraquinones. Normal phase TLC of the drug yielded 5 spots as against 6 spots yielded by reverse TLC.
Sahgal et al (2009) Methanolic extracts of Vitex negundo (Nochi), Coleus
amboinicus (Karpuravalli), Hylocereus polyrhyzus (Dragon fruit) and Andrographis paniculata (Hempedu Bumi) were evaluated in an effort to identify antibacterial compounds against clinical isolates of MethicillinResistant Staphylococcus aureus (MRSA). Coleus amboinicus crude methanolic (CACM) leaves extract inhibited MRSA growth completely at lower concentration compared to other extracts. This is the first time that the CACM leaves extract was tested and reported to be effective against MRSA. Hence, CACM leaves extract was chosen for determination of minimum inhibitory concentration (MIC), growth profile of MRSA, bioautographic and toxicity test against brine shrimp. The MIC value of
CACM leaves extract against MRSA was 200g.ml-1. CACM leaves extract resulted complete inhibition growth of the MRSA in growth profile and bioautography assay. The CACM leaves extract tested showed cytotoxicity activity against the Artemia salina. Therefore, CACM leaves extract could be used as a potential antibacterial agent from the natural plant sources for the treatment of MRSA infection.
M. N. Ibrahim et al (2008) Fresh Andrographis paniculata was dried by a
thin layer drying technique using hot air at a temperature of 55C and velocity of 1.0 m/s. The dried sample was then ground and kept in air-tight glass bottles at three selected storage conditions (52C; 252C with 60% 5% RH; 302C with (60% 5%RH) for three months period. The contents of the active compound andrographolide present in the dried Andrographis paniculata at the beginning and at the end of each month during the storage period were determined by using the HPLC. It was found that there was no significant reduction of the active compound andrographolide for all of the selected storage conditions. Storage at ambient conditions (30 2C, 60 5% RH) was also able to maintain andrographolide content in Andrographis paniculata during the three months period. This would enhance the production and supply of Andrographis paniculata as raw material with ease, without requiring the use of cooling equipment for storage purpose.
Johnson Marimuthu et al (2007) The present study was aimed at the
development of a somoclonal variant and isozyme marker for Phyllanthus amarus Schum & Thonn using inter-nodal segments and the enzyme peroxidase. Maximum callus proliferation was obtained on Murashige and Skoogs medium supplemented (MS) with 1.0 mg/l of 2,4Dichlorophenoxyacetic acid. Three weeks-old pale yellowish white semifriable callus was used for organogenesis; the maximum percentage of multiple shoot formation (85%) was achieved after 4 weeks when callus
was cultured on Murashige and Skoogs medium fortified with 1.0 mg/l of 6-Benzylaminopurine. The multiple shootlet formations were also achieved in the presence of the same concentration. The maximum formation of rootlets was observed on MS medium augmented with 1.0 mg/l of Kinetin and 0.5 mg/l of Naphthalene acetic acid. The banding pattern and phytochemical constituent differences were observed between mother plants, directly regenerated via nodal segments, calli, and calli mediated plants. The calli mediated somoclonal variation was confirmed through isozyme (peroxidase) and phytochemical analysis. The isoperoxidase banding profile showed a difference in calli and calli mediated plants. The phytochemical study confirmed the presence of more alkaloids, saponins, tannins and others from calli and calli mediated shoots and roots. Hence the isozyme banding patterns can be used as molecular markers in future plant breedings or genetic improvement programmes.
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SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

Specification Andrographolide 5.0%

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

Not more than 2.0 Appendix 2.1.3

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

Anti-fertility effect of Andrographis Paniculata (AP) was observed in mice under

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

Anti -allergic activity

antihepatotoxicactivity in vitro and in vivo (4554). Intraperitoneal administration

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

Ant malarial activity

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)

SHAILENDRA SINGH RAJPUT

RAJIV GANDHI INSTITUTE OF PHARMACY SHERGANJ, SATNA(M.P.)