Abstract:
The invention provides a novel herbal preparation useful in treatment of allergy. Formulation(s) comprises of plant preparation or active component of plant extract together with conventional additives to form the oral dosage forms, which includes tablets, capsules and liquid dosage forms viz. syrup and suspension as anti-allergic. Formulation(s) comprises of  Tinospora cardifolia, Piper longum, Albizia lebbeck  and  Curcuma amada . The formulation can be used as solid or liquid or encapsulated as a soft/hard gelatin capsule for easy consumption as solid or liquid dosage forms.

Description:
FIELD OF THE INVENTION 
     The present invention relates to novel herbal formulation(s) useful as anti-allergens. 
     BACKGROUND AND PRIOR ART OF THE INVENTION 
     Stress is known to alter the physiological homeostasis of the organism and results in the break down of the integrated adaptational processes. In response to extreme environmental demands various endocrinal and visceral responses occur naturally in a variety of experimental situations. For example, changes in plasma corticosterone and gastric mucosal integrity are widely reported during stress and both peripheral and central mechanisms seemingly regulate these changes. Much interest has recently been generated on the immunological changes during stress with reports indicating that the immune status of the organism is actually modified by experimental stressors. The central nervous system, besides being crucial for stress, also regulates immune function and studies show that common neural substrates like the hypothalamus are clearly involved in such central nervous system-immune system interactions. 
     Neuro-pharmacological data have shown that complex neurochemical mechanisms regulates stress responses and transmitters like gamma-aminobutyric acid (GABA) and endogenous opiates are crucially involved. For example, benzodiazepenes (BZD), which modulate GABA, and opioid antagonists, modify several stress responses like gastric ulcer formation and plasma corticosterone. Evidence indicates that central nervous system innervation of lymphoid tissue is possible. Lymphocytes bear receptors for several hormones and neurotransmitters and pharmacological alterations in neural activity influence immunocompetence. (Ray 1991) 
     Scientists now use the word anaphylaxis to mean any immune reaction of this type, even if it is not serious. But most doctors use it to mean life-threatening rapid allergic reaction. Unfortunately this kind of harmful immunization happens to a few of us not just from injections but also from ordinary foods such as nuts. Our immune system is there to protect us from infection, goes wrong and harms or even kills us. In recent years, there has been an upsurge in the clinical use of indigenous drugs. Such herbal plants, originally used in the traditional system of medicine, are now being effectively tried in a variety of path physiological states. Non-specific mechanisms like restoration of normal physiological milieu and generalized increase in resistance against infections are proposed and the role of the immune system in these drug effects is suggested. (Sen P et al., 1992). 
     OBJECT OF THE INVENTION 
     The main object of the present invention is to provide a novel Anti-allergic herbal formulation(s) in various dosage forms viz; tablet, capsule and ointment form for easy consumption. 
     Another objective of the present invention is to prepare herbal dosage form that improves in the treatment of allergy. 
     SUMMARY OF THE INVENTION 
     Accordingly, the present invention provides a novel of the invention of an herbal formulation(s) obtained from plants having the property of stimulant and resistant to various pathogens and acts as an immunomodulation as a tablet, injection and as a liquid formulation(s). 
     The herbal synergistic formulation has anti-allergy properties and comprises extracts of  Tinospora cardifolia, Piper longum, Albizia lebbeck  and  Curcunia amada.    
     In an embodiment the extracts/juice of the plants are mixed in the ratio viz  Tinospora cardifolia  (2-3 wt %),  Piper longum  (0.5-2 wt %),  Albizzia lebbeck  (3-5 wt %), and  Curcuma amada  (3-5 wt %) and balance being conventional additives. 
     In another embodiment, the plant used is  Tinospora cardifolia.    
     In another embodiment, the plant used is  Piper longum.    
     In another embodiment, the plant used is  Albizia lebbeck.    
     In still another embodiment, the plant used is  Curcuma amada.    
     In another embodiment, composition is used a syrup or suspension. 
     In another embodiment, the formulation is also used as anti-oxidant and nerves relaxant. 
     In still another embodiment, the formulation is also used to treat allergic related diseases, blood purifier, anti-periodic and externally applied sprain and wound. 
     In another embodiment, the formulation is also used as a blood purifier and for jaundice treatment. 
     In yet another embodiment, the said formulation has specific gravity ranging between 0.972-1.405 and a refractive index ranging between 1.5263-1.6812. 
     In another embodiment, the formulation at a dose of 400 mg/kg did not show any abnormality of general condition. 
     In another embodiment, the formulation form at a dose ranging from 100-200 mg/kg shown a 22.70-80.04% protection in hypoxia time and works as a stress relaxant. 
     In another embodiment, the formulation form at a dose ranging from 100-200 mg/kg shown a 24.18-82.83% protection in swimming endurance and works as a stress relaxant. 
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     Accordingly the present invention provides a novel Anti-allergic herbal formulation(s) in various dosage forms viz; tablet, capsule and ointment form for easy consumption. The herbal formulation comprising of  Tinospora cardifolia, Piper longum, Albizia lebbeck  and  Curcuma amada. Tinospora cardifolia  extract brown in color. It is used as nourishing food. It was shown that it produce a significant improvement in general ability and behavioural pattern. 
       Tinospora cordifolia  Family. Menisperniaceae 
     
         
         Botanical Descriptions: A large glabrous climbing shrub. Stems rather succulent with long filiform, aerial roots arising from branches. Bark; warty, papery thin, creamy white or grey brown. Peels off easily. Wood, soft, perforated. Leaves; membranous, cordate with broad sinus. Pointed at the tip. Flowers; unisexual and greenish, in long clusters. Seeds; curved. Drupes; ovoid, succulent, lustrous, red, pea sized. Fruits; fleshy, one seeded. It is found throughout tropical India, ascending to an altitude of 300 m. 
         Medicinal Uses: Useful in bilious fever, rheumatism, general debility, seminal weakness, splenic diseases and urinary affections. Fresh plant is considered more efficacious. It is mostly used for preparing a kind of starch known as  Guduchi satva  or  Sat giloe.    
         Phytochemistry: Sesquiterpene tinocordifolin, sesquiterpene glucoside tinocordifolioside, tinosponone, tinocordioside, cordioside, furanoid diterpenes, a new clerodane furano-diterpene viz. columbin, tinosporaside, an immunologically active arabinogalactan, two phytoecdyyones viz., ecdysterone and makisterone and several glycosides isolated as polyacetates. Other alkaloids viz., jatrorrhizine, palmatine, berberine, tembeterine, phenylpropene disaccharides cordifolioside A, B and C, choline, tinosporic acid, tinosporal, tinosporon, 20-β-hydroxyecdysone, palmatoside C and F, cordifolisides D and E, diterpenoid furanolactones. 
         Pharmacology: The water and ethanolic extract inhibited the cyclophosphamide induced immunosupression. Aqueous extract of the stem showed anti-inflammatory, analgestic and antipyretic properties in rats. In clinical studies, it also showed immunosuppressive effect in obstructive jaundice patients, antioxidant activity and amelioration of cylcophosphamide-induced toxicity.
 
 Piper longum  L. Family: Piperaceae
 
         Botanical Description: A slender, aromatic climber with perennial woody roots; stems jointed, creeping, young shoots downy. Leaves simple, alternate, 5-12 cm long and 3-6 cm wide, glabrous, ovate base cord ate with broad rounded lobes, apex sub acute, and margins entire. Flowers minute on unisexual, axillary, cylindrical spikes; green at first, turning yellow, up to 5 cm long; male spikes longer than female. Fruits ovoid, yellowish orange turning dark red to blackish, sunk in fleshy spikes 2.5-3.8 cm long. 
         Pharmacology: The dried roots, as well as the immature and mature fruits, are used extensively, alone and in combination with other plant drug to treat a broad range of ailments in traditional Indian medicine. The dried roots and thicker stems, known commercially as piplamul, are an important drug in the ayurvedic and unani systems. The roots and fruits are used to treat dysentery and leucoderma, as a cholagogue for treating bileduct and gallbladder obstruction, and as a counter—irritant and analgesic for relieving muscularpains and inflammations. A decoction of dried immature fruit and root, or the powdered fruits mixed with honey, is used to treat chronic bronchitis, cough and cold. An infusion of the powdered fruits is given to women after childbirth to check bleeding and fever. It is as important ingredient in medicated oil used externally for sciatica and paraplegia.
 
 Albizia lebbeck  Family: Mimosoideae
 
         Botanical description: It is common throughout India, from the plains up to 900 m elevations in the Himalayas. It is a large, unarmed, decidous tree to about 20 m tall with a spreading crown; bark pale; young shoots glabrous. Leaves bipinnate; rachis with a gland near the base of the petiole and one below uppermost pairs of pinnae; pinnae usually 2-3 pairs, 7-12 cm long; leaflets 5-9 pairs, 2.5-4.5 cm long and 1.6-2 cm wide, with glands between their bases, elliptic-oblong or obovate-oblong, unequal-sided, retuse or obtuse at apex, base obliquely rounded or truncate; petiolules very short. Flowers fragrant, white to greenish-yellow, borne in globose umbellate heads 2-3.8 cm in diameter; peduncles 3.8-7.5 cm long, solitary or 2-4 together from the axils of the upper leaves, pedicels 2.5-3 mm long; calyx 4 mm long, pubescent, teeth short, deltoid; corolla 1 cm long, tube glabrous, lobes 2.5 mm long, triangular, acute, pubescent outside; stamens longer than the corolla. Fruits (pods) 10-30 cm long and 2-4.5 cm wide, linear-oblong, bluntly pointed, thin green turning straw-coloured on maturity, reticulately veined above the seeds, smooth, shining; seeds 4-12, pale brown, ellipsoid-oblong, compressed. Flowers from April to June and fruits mature in December in northern and central India; in southern India flowering occurs earlier, from January to April. In central India trees remains leafless for a month or more between March and June. 
         Medicinal uses: In Ayurveda the astringent root is used to treat hemicrania; the acrid bark is reportedly used to treat diseases of the blood, leucoderma, itching, skin diseases, piles, inflammation, and bronchitis; the leaves are used to treat opthalmia, and the flowers for asthma. In unani medicine the root is used to treat ophthalmia; the bark is regarded as anthelmintic and used to relieve toothache and to strengthens the gums and teeth, and to treat leprosy, deafness, boils, scabies, syphilis and paralysis; the leaves are reportedly useful for treating night blindness; and the seeds are used to treat gonorrhoea and tuberculous glands, their oil applied locally for leucoderma. The flower are used as a cooling medicine and as an external application to relieve boils, skin eruption and swelling the seed oil is used externally to promote healing of lesions in leprosy among the Irulars of Tamilnadu. 
         Phytochemistry: A new acyclic ester isolated and characterized as heneicos-7 (2)-enyl-24-hydroxytetracos-10 (2)-enoate; lupeol, oleanolic acid, docosanoic acid and β-sitosterol also was isolated ( Ind J Pharm Sci,  1991,53,24). Echinocystic acid β-sitosterol identified in bark and seeds ( Ind J Appl. Chem,  1969,32,73 ; Chem Abstr  1971,75,160352). Mature leaves contained ketoacids including phosphoenol pyruvate, glyoxalate, oxaloacetate and α-oxoglutarate ( Plant Biochem J,  1977,4,34; Chem Abstr 1977,17,148762s); vicenin-2, reynoutrin, rutin, myricitrin and robinin from leaves ( Shoyakugaku Zasshi;  1977, 31, 172 ; Chem Abstr  1978,88,14897h). 
         Pharmacology: Bark and flowers decoction protected guinea pig against histamine and acetlycholine-induced bronchospam. Chronic treatment with bark decoction also protected sensitized guinea pigs against antigen challenge. Drug showed anti-asthmatic and anti-anaphylactic activities due to inhibition of phenomenon of sensitization ( Ind J Pharmacol,  1977,9,189). Saponin showed β-haemolysis against buffalo and sheep blood and α-haemolysis against human blood. Purified saponin showed antifungal activity against marcophomnina phaseolina (mic 32.8 μg), stemphilum species and  fusarium  solani ( Pak Vet J  1990,10,146 ; Chemi Abstr  1992,116,102686j). The plant has been showed to poses antidirrhoeal and antifertility activities. It posses anti anaphylactic and anticonvulsive properties. The leaves are reportedly useful for treating night blindness; and the seeds are used to treat gonorrhoea and tuberculous glands, their oil applied locally for leucoderma.
 
 Curcuma amada  Family: Zingiberaceae
 
         Botanical description: A tall herb, rootstock large, ovoid, with cylindrical tubers that are bright yellow or orange inside. Leaves very large, in tufts up to 1.2 m long including petioles, blade up to 50 cm long and 8 cm wide, oblong lanceolate, tapering to the base; petiole about as long as leaf blade. Flowers are pale yellow. 
         Pharmacology: The dried, powdered rhizome is the source of turmeric, widely used in Indian cuisine as well as in traditional medicine. Turmeric is considered to have strong antiseptic properties and is used to treat poisonous affections, ulcers and wounds. In Ayurveda, is considered to be alterative, antiperiodic, germicidal, carminative, stimulant, tonic and vermifuge. It is used to treat diabetes, eye diseases, ulcers, oedema, anaemia, anorexia, leprosy, scrofula; its paste, sometimes combined with that of neem leaves, is used to cure ring worm, itching, eczema and other parasitic skin diseases, and in the treatment of chicken pox and small pox. In unani medicine it is considered carminative, mutant and diuretic and useful for treating jaundice and other liver disorders. An essential oil obtained from the rhizome is used as a carminative, stoma chic and tonic. A paste made of the flowers is used externally to treat ringworm and other parasitic skin diseases, as well as gonorrhoea. 
         Medicinal uses: It is used to treat diabetes, eye diseases, ulcers, oedema, anaemia, anorexia, leprosy, scrofula; its paste, sometimes combined with that of neem leaves, is used to cure ring worm, itching, eczema and other parasitic skin diseases, and in the treatment of chicken pox and small pox. 
       
    
     The present invention provides a novel of the invention of an herbal formulation(s) obtained from plants having the property of stimulant and resistant to various pathogens and acts as an immunomodulation as a tablet, injection and as a liquid formulation(s). 
     The herbal synergistic formulation has anti-allergy properties and comprises extracts of  Tinospora cardifolia, Piper longum, Albizia lebbeck  and  Curcuma amada . The extracts/juice of the plants are mixed in the ratio viz  Tinospora cardifolia  (2-3 wt %),  Piper longum  (0.5-2 wt %),  Albizzia lebbeck  (3-5 wt %), and  Curcuma amada  (3-5 wt %) and balance being conventional additives. 
     In another embodiment, the plant used is  Tinospora cardifolia.    
     In another embodiment, the plant used is  Piper longum.    
     In another embodiment, the plant used is  Albizia lebbeck.    
     In still another embodiment, the plant used is  Curcuma amada.    
     In another embodiment, composition is used a syrup or suspension. 
     The formulation is also used as anti-oxidant and nerves relaxant. The formulation can also be used to treat allergic related diseases, blood purifier, anti-periodic and externally applied sprain and wound. The formulation is also used as a blood purifier and for jaundice treatment. 
     The formulation has specific gravity ranging between 0.972-1.405 and a refractive index ranging between 1.5263-1.6812. At a dose of 400 mg/kg did not show any abnormality of general condition. The formulation form at a dose ranging from 100-200 mg/kg shown a 22.70-80.04% protection in hypoxia time and works as a stress relaxant. 
     The formulation form at a dose ranging from 100-200 mg/kg shows 24.18-82.83% protection in swimming endurance and works as a stress relaxant. 
     The invention is further illustrated by the following non-limiting examples. 
     Formulation 1 (F1) 
     
       
         
               
               
               
               
             
               
               
               
             
           
               
                   
                   
               
             
             
               
                   
                 
                   Tinospora cardifolia 
                 
                 3 
                 wt. % 
               
               
                   
                 Lactose 
                 66.7 
                 g 
               
               
                   
                 Starch 
                 10 
                 g 
               
             
          
           
               
                   
                 Water 
                 q.s. to make 100 ml 
               
               
                   
                   
               
             
          
         
       
     
     Dry mature seed of  Tinospora cardifolia  are washed in purified distilled water, and were immersed in hot water (30-60° C.) for 2-3 hr. Mix the plant constituents and filter the solution and add specified quantity of starch and heat until the starch dissolves and then cool and make up the volume with required amount of water to make 100 ml. Oral dosage form has been described in detail giving the formula of the ingredients along with the method and mode of usage of the standardized formulation. Kindly refer table I and II. 
     Formulation 2 (F2) 
     
       
         
               
               
               
               
             
               
               
               
             
           
               
                   
                   
               
             
             
               
                   
                 
                   Piper longum 
                 
                 1 
                 wt. % 
               
               
                   
                 Lactose 
                 66.7 
                 g 
               
               
                   
                 Starch 
                 10.0 
                 g 
               
             
          
           
               
                   
                 Water 
                 q.s. to make 100 ml 
               
               
                   
                   
               
             
          
         
       
     
     Dry mature seed of  Piper lonigum  are washed in purified distilled water, and were immersed in hot water (30-60° C.) for 2-3 hr. Mix the plant constituents and filter the solution and add specified quantity of starch and heat until the starch dissolves and then cool and make up the volume with required amount of water to make 100 ml. Oral dosage form has been described in detail giving the formula of the ingredients along with the method and mode of usage of the standardized formulation. Kindly refer table I and II. 
     Formulation 3 (F3) 
     
       
         
               
               
               
               
             
               
               
               
             
           
               
                   
                   
               
             
             
               
                   
                 
                   Albizia lebbeck 
                 
                 4 
                 wt. % 
               
               
                   
                 Lactose 
                 66.7 
                 g 
               
               
                   
                 Starch 
                 10.0 
                 g 
               
             
          
           
               
                   
                 Water 
                 q.s. to make 100 ml 
               
               
                   
                   
               
             
          
         
       
     
     Dry mature seed of  Albizia lebbeck  are washed in purified distilled water, and were immersed in hot water (30-60° C.) for 2-3 hr. Mix the plant constituents and filter the solution and add specified quantity of starch and heat until the starch dissolves and then cool and make up the volume with required amount of water to make 100 ml. Oral dosage form has been described in detail giving the formula of the ingredients along with the method and mode of usage of the standardized formulation. Kindly refer table I and II. 
     Formulation 4 (F4) 
     
       
         
               
               
               
               
             
               
               
               
             
           
               
                   
                   
               
             
             
               
                   
                 
                   Curcuma amada 
                 
                 5 
                 wt % 
               
               
                   
                 Lactose 
                 66.7 
                 g 
               
               
                   
                 Starch 
                 10 
                 g 
               
             
          
           
               
                   
                 Water 
                 q.s. to make 100 ml 
               
               
                   
                   
               
             
          
         
       
     
     Dry mature seed of  Curcuma amada  are washed in purified distilled water, and were immersed in hot water (30-60° C.) for 2-3 hr. Mix the plant constituents and filter the solution and add specified quantity of starch and heat until the starch dissolves and then cool and make up the volume with required amount of water to make 100 ml. Oral dosage form has been described in detail giving the formula of the ingredients along with the method and mode of usage of the standardized formulation (Tables I and II) 
     Formulation 5 (F5) 
     
       
         
               
               
               
               
             
               
               
               
             
           
               
                   
                   
               
             
             
               
                   
                 
                   Tinospora cardifolia 
                 
                 3 
                 wt. % 
               
               
                   
                 
                   Piper longum 
                 
                 1 
                 wt. % 
               
               
                   
                 Lactose 
                 63.7 
                 g 
               
               
                   
                 Starch 
                 10 
                 g 
               
             
          
           
               
                   
                 Water 
                 q.s. to make 100 ml 
               
               
                   
                   
               
             
          
         
       
     
     Dry mature seed of  Tinospora cardifolia  and  Piper longum  are washed in purified distilled water, and immersed in hot water (30-60° C.) for 2-3 hr. Mix the plant constituents and filter the solution and add specified quantity of starch and heat until the starch dissolves and then cool and make up the volume with required amount of water to make 100 ml. Oral dosage form has been described in detail giving the formula of the ingredients along with the method and mode of usage of the standardized formulation (Tables I and II). 
     Formulation 6 (F6) 
     
       
         
               
               
               
               
             
               
               
               
             
           
               
                   
                   
               
             
             
               
                   
                 
                   Tinospora cardifolia 
                 
                 3 
                 wt. % 
               
               
                   
                 
                   Piper longum 
                 
                 1 
                 wt. % 
               
               
                   
                 
                   Albizia lebbeck 
                 
                 3 
                 wt. % 
               
               
                   
                 Lactose 
                 60.7 
                 g 
               
               
                   
                 Starch 
                 10 
                 g 
               
             
          
           
               
                   
                 Water 
                 q.s. to make 100 ml 
               
               
                   
                   
               
             
          
         
       
     
     Dry mature seed of  Tinospora cardifolia, Piper longum  and  Albizia lebbeck  are washed in purified distilled water, and immersed in hot water (30-60° C.) for 2-3 hr. The plant constituents were mixed and the solution filtered and specified quantity of starch added and heated till starch dissolves and then cooled and volume made up with required amount of water to make 100 ml. Oral dosage form has been described in detail giving the formula of the ingredients along with the method and mode of usage of the standardized formulation. Kindly refer table I and II. 
     Formulation 7(F7) 
     
       
         
               
               
               
               
             
               
               
               
             
           
               
                   
                   
               
             
             
               
                   
                 
                   Tinospora cardifolia 
                 
                 3 
                 wt. % 
               
               
                   
                 
                   Piper longum 
                 
                 1 
                 wt. % 
               
               
                   
                 
                   Albizia lebbeck 
                 
                 4 
                 wt. % 
               
               
                   
                 
                   Curcuma amada 
                 
                 3 
                 wt. % 
               
               
                   
                 Lactose 
                 56.7 
                 g 
               
               
                   
                 Starch 
                 10 
                 g 
               
             
          
           
               
                   
                 Water 
                 q.s. to make 100 ml 
               
               
                   
                   
               
             
          
         
       
     
     Dry mature seed of  Tinospora cardifolia, Piper longum, Albizia lebbeck  and  Curcuma amada  are washed in purified distilled water, and were immersed in hot water (30-60° C.) for 2-3 hr. Mix the plant constituents and filter the solution and add specified quantity of starch and heat until the starch dissolves and then cool and make up the volume with required amount of water to make 100 ml. Oral dosage form has been described in detail giving the formula of the ingredients along with the method and mode of usage of the standardized formulation. Kindly refer table I and II. 
     
       
         
               
             
               
               
               
               
               
             
           
               
                 TABLE I 
               
             
             
               
                   
               
               
                 Effect of formulation on hypoxia and 
               
               
                 swimming performance time in mice. 
               
             
          
           
               
                   
                   
                 Dose 
                   
                 Swimming 
               
               
                 S. No 
                 Treatment 
                 (mg/kg, p.o) 
                 Hypoxia time 
                 endurance 
               
               
                   
               
               
                 1. 
                 Control + 
                 — 
                 23.34 ± 1.15 
                 238.21 ± 25.11 
               
               
                   
                 Stress 
               
               
                 2. 
                 F1 
                 200 
                 24.34 ± 1.55 
                 241.21 ± 27.41 
               
               
                 3. 
                 F2 
                 200 
                 22.54 ± 2.11 
                 239.45 ± 25.05 
               
               
                 4. 
                 F3 
                 200 
                 26.01 ± 2.28 
                 245.52 ± 26.74 
               
               
                 5. 
                 F4 
                 200 
                 27.89 ± 2.15 
                 246.32 ± 25.53 
               
               
                 6. 
                 F5 
                 200 
                 27.01 ± 2.58 
                 248.14 ± 26.85 
               
               
                 7. 
                 F6 
                 200 
                 35.23 ± 2.95 a   
                 369.52 ± 26.74 a   
               
               
                 8. 
                 F7 
                 200 
                 45.89 ± 3.05 b   
                 435.54 ± 28.85 b   
               
               
                   
               
               
                 Values are mean ± S.E.M. 
               
               
                 P:  a &lt;0.01 and  b &lt;0.001 compared to control + stress group. 
               
               
                 NOTE: 
               
               
                 No mortality was found in any of the treated group. 
               
             
          
         
       
     
     No gross abnormality in behavior was observed in the animal exposed with herbal preparation. The formulation (F1) contains  Tinospora cardifolia  (3%) with balance being conventional additives. The formulation (F2) contains  Piper longum  (1%) with balance being conventional additives. The formulation (F3) contains  Albizia lebbeck . (4%) with balance being conventional additives. The formulation (F4) contains  Curcuma amada  (3%) with balance being conventional additives. The formulation (F5) contains  Tinospora cardifolia  (3%) and  Piper longum  (1%) with balance being conventional additives. The formulation (F6) contains  Tinospora cardifolia  (3%),  Piper longum  (1%) and  Albizia lebbeck  (3%) with balance being conventional additives. The formulation (F7) contains  Tinospora cardifolia  (3%),  Piper longum  (1%),  Albizia lebbeck  (4%) and  Curcuma amada  (3%) with balance being conventional additives 
     The results showed in table 1 that formulations F7 showed significant hypoxia time when compared with control+stress and formulation F1-F6. The value of formulation F7 hypoxia time has significantly increased and swimming endurance value is decreased which leads to stress relaxation. 
     Immunological Assay 
     
         
         Hypoxia time: The animals were placed in an empty glass jar of 300 mL capacity attached with an electronic watch; the jars were made air tight with greased glass stoppers and the time until the onset of convulsion was recorded (Singh B et al 2001). 
         Swimming performance time: The animals were allowed to swim inside a Perspex glass beaker (30 cm high with 20 cm diameter containing water up to 25 cm high) maintained at 26°±1° C. with a continuous air current from the bottom. The end point of swimming endurance was taken as when the mice remained at the bottom for more than 10 s (Singh B et al 2001). 
       
    
     
       
         
               
             
               
               
               
               
             
           
               
                 TABLE II 
               
             
             
               
                   
               
               
                 Effect of formulation on mast cell degranulation in rats. 
               
             
          
           
               
                   
                   
                 Dose 
                 Histamine released 
               
               
                 S. No 
                 Treatment 
                 (mg/kg, p.o) 
                 (μg/ml) 
               
               
                   
               
               
                 1. 
                 Control 
                 — 
                 0.899 ± 0.05 
               
               
                 2. 
                 F1 
                 200 
                 0.895 ± 0.04 
               
               
                 3. 
                 F2 
                 200 
                 0.886 ± 0.06 
               
               
                 4. 
                 F3 
                 200 
                 0.892 ± 0.05 
               
               
                 5. 
                 F4 
                 200 
                 0.879 ± 0.05 
               
               
                 6. 
                 F5 
                 200 
                 0.885 ± 0.02 
               
               
                 7. 
                 F6 
                 200 
                 0.671 ± 0.02 a   
               
               
                 8. 
                 F7 
                 200 
                 0.254 ± 0.05 c   
               
               
                 9. 
                 Avil 
                 25 
                 0.345 ± 0.04 b   
               
               
                   
               
               
                 Values are mean ± S.E.M. 
               
               
                 P:  a &lt;0.05,  b &lt;0.01 and  c &lt;0.001 compared to control group. 
               
               
                 NOTE: 
               
               
                 No mortality was found in any of the treated group. 
               
             
          
         
       
     
     No gross abnormality in behavior was observed in the animal exposed with herbal preparation. 
     The formulation (F1) contains  Tinospora cardifolia  (3%) with balance being conventional additives. The formulation (F2) contains  Piper longum  (1%) with balance being conventional additives. The formulation (F3) contains  Albizia lebbeck . (4%) with balance being conventional additives. The formulation (F4) contains  Curcuma amada  (3%) with balance being conventional additives. The formulation (F5) contains  Tinospora cardifolia  (3%) and  Piper longum  (1%) with balance being conventional additives. The formulation (F6) contains  Tinospora cardifolia  (3%),  Piper longum  (1%) and  Albizia lebbeck  (3%) with balance being conventional additives. The formulation (F7) contains  Tinospora cardifolia  (3%),  Piper longum  (1%),  Albizia lebbeck  (4%) and  Curcuma amada  (3%) with balance being conventional additives. 
     The results showed in table 2 formulations F7 showed significant effect when compared with control. The release of histamine is tremendously decreased which shows the anti allergic activity. The formulation F7 is more significant than that of standard drug Avil. 
     Disadvantages of Avil are Dizziness and Drowsiness. 
     
         
         Mast cell degranulation: Tyrode solution (10 ml) containing 5 units/ml of heparin was injected in the peritoneal cavity of sensitized male rats lightly anaesthetized with ether. After a gentle abdominal massage for about 30 to 45 sec, the mast cell rich peritoneal fluid was collected over ice and centrifuged at 2000 rpm for 5 min. The cells were washed thrice with chilled Tyrode and resuspended in about 1 ml of Tyrode solution. The peritoneal fluid was obtained from 10 rats and the final volume of the fluid was pooled and used for the studies (Nair A M et al, 1997). 
       
    
     
       
         
               
             
               
               
               
               
               
             
               
             
               
               
               
               
               
             
               
             
               
               
               
               
               
             
               
             
               
               
               
               
               
             
           
               
                 TABLE III 
               
             
             
               
                   
               
               
                 Effect of formulation (F7) on lipid peroxidation, superoxide 
               
               
                 dismutase and catalase activities in tissues. 
               
             
          
           
               
                 S. No 
                 Treatment 
                 Liver 
                 Kidney 
                 Heart 
               
               
                   
               
             
          
           
               
                 Lipidperoxidation(LPO) 
               
             
          
           
               
                 1. 
                 Control 
                 32.2 ± 1.5 
                  25.1 ± 1.3 
                 17.7 ± 0.5 
               
               
                 2. 
                 F7 
                      18.5 ± 1.2 c   
                      13.65 ± 2.7 c   
                      21.5 ± 2.1 a   
               
             
          
           
               
                 Superoxide dismutase(SOD) 
               
             
          
           
               
                 1. 
                 Control 
                  9.2 ± 0.6 
                  5.8 ± 0.9 
                  4.3 ± 0.5 
               
               
                 2. 
                 F7 
                    2.6 ± 0.8 c   
                     3.6 ± 0.2 b   
                    4.9 ± 0.4 a   
               
             
          
           
               
                 Catalase activity 
               
             
          
           
               
                 1. 
                 Control 
                 96.3 ± 1.8 
                 126.5 ± 1.9 
                 92.5 ± 2.5 
               
               
                 2. 
                 F7 
                 185.6 ± 7.1 c       
                      256.9 ± 8.3 c   
                 132.2 ± 3.4 c       
               
               
                   
               
               
                 Values are mean ± S.E.M. 
               
               
                 P:  a &lt;0.05,  b &lt;0.01 and  c &lt;0.001 compared to respective control group. 
               
               
                 NOTE: 
               
               
                 No mortality was found in any of the treated group. 
               
             
          
         
       
     
     No gross abnormality in behavior was observed in the animal exposed with herbal preparation. 
     The formulation (F7) contains  Tinospora cardifolia  (3%),  Piper longum  (1%),  Albizia lebbeck  (4%) and  Curcuma amada  (3%) with balance being conventional additives. 
     
       
         
               
             
               
               
               
               
               
             
           
               
                 TABLE IV 
               
             
             
               
                   
               
               
                 Effect of formulation(s) on relative 
               
               
                 mean ± SEM organ weights of rats. 
               
             
          
           
               
                 Treatment 
                   
                   
                   
                   
               
               
                 group 
                 Body weight (g) 
                 Kidney (g) 
                 Liver (g) 
                 Spleen (g) 
               
               
                   
               
               
                 Control 
                 182.5 ± 9.3 
                 0.94 ± 0.04 
                 5.92 ± 0.65 
                 0.61 ± 0.07 
               
               
                 F (5) 
                 171.3 ± 8.6 
                 0.87 ± 0.05 
                 5.68 ± 0.58 
                 0.68 ± 0.05 
               
               
                 F (6) 
                 178.1 ± 9.7 
                 0.92 ± 0.07 
                 5.82 ± 0.61 
                 0.70 ± 0.08 
               
               
                 F (7) 
                 185.4 ± 8.4 
                 0.85 ± 0.09 
                 5.13 ± 0.69 
                 0.65 ± 0.06 
               
               
                   
               
             
          
         
       
     
     The formulation (F5) contains  Tinospora cardifolia  (3%) and  Piper longum  (1%) with balance being conventional additives. The formulation (F6) contains  Tinospora cardifolia  (3%),  Piper longum  (1%) and  Albizia lebbeck  (3%) with balance being conventional additives. The formulation (F7) contains  Tinospora cardifolia  (3%),  Piper longum  (1%),  Albizia lebbeck  (4%) and  Curcuma amada  (3%) with balance being conventional additives. 
     The results of the Table IV shows there is no significant changes in body weight of various vital organs in the body in toxicity studies. 
     The formulation F5, F6 and F7 is highly effective (Table I) and it is safe. 
     Note: No mortality/gross abnormality was observed in the animals during the treatment of formulations (F5, F6 and F7).