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 Table of Contents  
ORIGINAL ARTICLE
Year : 2016  |  Volume : 1  |  Issue : 2  |  Page : 133-138

Pharmaceutico-physicochemical validation of Mamajjaka Ghana tablet, a potent antidiabetic medicine


1 Department of Rasa Shastra and Bhaishajya Kalpana, IPGT and RA, Gujarat Ayurved University, Jamnagar, Gujarat, India
2 Department of Dravyaguna, IPGT and RA, Gujarat Ayurved University, Jamnagar, Gujarat, India

Date of Submission21-Jun-2016
Date of Acceptance01-Dec-2016
Date of Web Publication19-Dec-2016

Correspondence Address:
Ashish Verma
PG Boys Hostel, Room No. 64, IPGT and RA, Gujarat Ayurveda University, Jamnagar, Gujarat
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/2468-838X.196106

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  Abstract 

Background: Mamajjaka Ghana Vati is a well-recognized herbal medicine for the treatment of Madhumeha. Instead of its attribute in Madhumeha, it was also therapeutically indicated for antipyretic, anthelmintic, hepatoprotective, and anti-inflammatory activities.
Aims and Objective: The objective of the present study was to evaluate the significance of large-scale production of Mamajjaka Ghana tablet and its quality control aspect with a view of its wide range of applicability in therapeutics and ayurvedic pharmaceutical industry.
Materials ad Methods: Mamajjaka Ghana tablet was prepared in four batches and the findings were scientifically recorded. In this analytical study, organoleptic and physicochemical evaluations of Mamajjaka decoction, solid aqueous extract (Ghana), and tablet were carried out.
Results and Discussion: An average of 38.25 h were required for the preparation of Mamajjaka decoction. The average temperature maintained during the process was 95-100°C. The average yield of decoction was 168.75 L in all the four batches. An average of 11.67 kg of solid aqueous extract (Ghana) was obtained. A total of 46.68 kg of Ghana was obtained from all the four batches. An average of 97.93% of Mamajjaka Ghana tablet was obtained.
Conclusion: The data pertaining to large-scale production will be useful for ayurvedic pharmaceutical industry populace to maintain quality, uniformity of dosage, and to sustain the batch-to-batch variation during various pharmaceutical operations, and thus the whole process is validated for 65.28 kg of Mamajjaka Ghana tablet .

Keywords: Ghana, Mamajjaka, tablet, validation


How to cite this article:
Verma A, Janani H, Yadav S, Ghimire S, Patgiri B J, Prajapati P K. Pharmaceutico-physicochemical validation of Mamajjaka Ghana tablet, a potent antidiabetic medicine. BLDE Univ J Health Sci 2016;1:133-8

How to cite this URL:
Verma A, Janani H, Yadav S, Ghimire S, Patgiri B J, Prajapati P K. Pharmaceutico-physicochemical validation of Mamajjaka Ghana tablet, a potent antidiabetic medicine. BLDE Univ J Health Sci [serial online] 2016 [cited 2019 Aug 20];1:133-8. Available from: http://www.bldeujournalhs.in/text.asp?2016/1/2/133/196106

Extraction, as the term is used pharmaceutically, involves the separation of medicinally active portions of plant or animal tissues by using selective solvents in standard extraction procedures. The products so obtained from plants are liquids, semisolids, or powders intended only for oral or external use. These include classes of preparations known as decoctions, infusions, fluid extracts, tinctures, pilular (semisolid) extracts, and powdered extracts. The purposes of standardized extraction procedures for crude drugs are to attain the therapeutically desired portion and to eliminate the inert material by treatment with a selective solvent known as menstruum, a solvent, especially the one used in extracting compounds from plant and animal tissues and in preparing drugs.

The extract thus obtained may be ready for use as a medicinal agent in the form of tinctures or extracts and further processed in any dosage form such as tablets or capsules. All these products contain a complex mixture of many medicinal plant metabolites, such as alkaloids, glycosides, terpenoids, flavonoids, and lignans. To be used as a modern drug, an extract may be further processed through various techniques of fractionation to isolate individual chemical entities such as vincristine, vinblastine, hyoscyamine, hyoscine, pilocarpine, forskolin, and codeine. Thus, standardization of extraction procedures contributes significantly to the final quality of the herbal drug.

In Ayurveda classics, five different dosage forms are given, collectively called Panchavidha Kashaya Kalpana.[1] These are the basis of all the secondary Kalpanas such as Avaleha (semisolid dosage form), Sandhana (hydro-alcoholic preparation), and Sneha (fatty preparations). Instead of their valuable importance in ayurvedic pharmaceutics, they have some drawbacks such as nonavailability of crude drugs all the time, very short shelf life, and inconvenient taste and dose. To counteract these problems, Upakalpana and secondary Kalpana were developed using the basic Kalpana.[2] Another motto behind the development of Upakalpana seems to enhance the potency as well as the selectivity in the pharmacological actions.

Ghana (solid aqueous extract) is a concentrated dosage form, which is a modification of Kwatha (decoction) Kalpana to increase potency, shelf life, and also to minimize the dosage of drug. Ghana is nothing but the semisolidification or solidification of the liquid substances or the formulations which is also termed Rasakriya, Lehya, and Avaleha in ayurvedic classics.[3] It has not been mentioned in any of the classics as a separate formulation or process. Though Avaleha and Lehya are considered synonyms of Rasakriya by Acharya Sharangadhara, a slight variation in their pharmaceutical preparation can be recognized. These dosage forms are prepared by evaporating the water content of aqueous solutions (decoction) and then concentrated up to semisolid form to prepare Vati (pills) or up to dried form to prepare Ghana (aqueous extract), if necessary.

Mamajjaka (Enicostemma littorale Blume) is one of the famous traditional herbs used in Madhumeha, especially in Gujarat, Madhya Pradesh, and Rajasthan. It is used as a substitute for Chirayata (Swertia chirayita Roxb. Lyons).[4] Mamajjaka is mentioned in Shodhala Nighantu in Lakshmanadi Varga. It has the synonyms, namely, Nagajihva, Mamajjaka, Nahi, Tiksnapatra, Vitikshnika, Krimihrit, and Ksharakarma.[5] It has Tikta rasa (bitter taste), Laghu guna, Ushna veerya (hot in potency), and Katu Vipak, and mentioned as Kaphapittashamak (subsided all the three humors). It is therapeutically indicated in Amadosha (indigestion), Vibandha (constipation), Yakritdaurbalya (liver disorders), Krimi (worms), Raktavikara (blood disorders), Shotha (inflammation), Prameha (urinary disorders), Madhumeha (diabetes), Twagvikara (skin disease), Vishamajwara (intermittent fever), Medoroga (obesity), and Atisara (dysentery).[6] It is also indicated for fever, malaria, diarrhea, and worms.[7] Mamajjaka is widely grown in Saurashtra region of Gujarat and traditionally people also prepare pickle out of it.

Mamajjaka Ghana Vati[8] is traditionally known by the ayurvedic alliance for its therapeutic properties on Madhumeha. Today, more than twenty ayurvedic antidiabetic formulations are available in the market containing Mamajjaka as an ingredient.[9] Hence, it represents the importance of Mamajjaka as an ingredient into multiple herbal formulations and its wide range of applicability.

In the present scenario, ayurvedic pharmaceutical industry is increasing at a rapid pace over the years. Most of the industries are launching new and unique dosage forms which have a long shelf life, good elegance, and they are palatable, acceptable, and compatible with the expectations of customers. Palatability is the major setback in the case of decoction and powder. Tablet form has a wide range of applicability such as uniformity in disintegration and dissolution time. Except that palatability, shelf life and eases in administration are the other benefits of the tablet form. One more reason behind the selection of this study was that large-scale production and quality control of Mamajjaka Ghana tablet have not been taken for the study till date. Hence, it has been preferred for the present study.


  Materials and Methods Top


Identification and authentication

Dried whole plant of Mamajjaka (Enicostemma littorale blume), 320 kg, and Prakshepa (powder) Dravya (Mamajjaka, Katuki, Ativisha, and Pippali, each 4 kg in raw form) were procured from Department of Pharmacy, Gujarat Ayurved University, Jamnagar, and authenticated at Pharmacognosy laboratory, IPGT and RA, Jamnagar. All the pharmaceutical operations of Mamajjaka Ghana tablet were carried out at the Pharmacy Department, Gujarat Ayurved University, Jamnagar.

Preparation of Mamajjaka Ghana tablet

The whole procedure was divided into the following three parts:

  1. Preparation of Mamajjaka decoction
  2. Preparation of Mamajjaka aqueous extract (Ghana)
  3. Preparation of Mamajjaka Ghana tablet.


Coarse powder of dried whole plant of Mamajjaka was prepared using a pulveriser. Coarse powder of Mamajjaka was soaked overnight (12 h) with eight parts (640 L) of reverse osmosis (RO) water and finally reduced to 1/4th of its original volume and filtered through 4-folded clean cotton cloth to obtain decoction. Similarly, four batches of decoction were prepared [Table 1].
Table 1: Observations and results obtained during the preparation of Mamajjaka decoction


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The decoction was subjected to the process of reboiling with infrequent stirring over mild flame till the contents become semisolid. Semisolid aqueous extract (Ghana) thus obtained was collected and used for further processing. In a similar manner, four batches of Mamajjaka Ghana were prepared [Table 2].
Table 2: Observations and results obtained during the preparation of Mamajjaka Ghana


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Preparation of Mamajjaka Ghana tablet

Semisolid aqueous extract of Mamajjaka (moisture 20%) was mixed (triturated) with Prakshepa Dravya (powders) and solution of gum acacia in end runner. Uniform mass, thus obtained, was converted into small lumps and subjected for drying in an oven. Further, granules were prepared from the dried lumps, dried, and excipients were added and compressed into tablet form with the help of Single Rotary Tablet machine of 16 stations. Tablets were collected, weighed, and kept into pharmaceutical grade air-tight plastic containers. By adopting aforesaid method, four batches of Mamajjaka Ghana tablet were prepared [Table 3], [Table 4], [Table 5].
Table 3: The proportion of ingredients in Mamajjaka Ghana tablet


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Table 4: Results obtained during the granulation and mixing of excipients


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Table 5: Results obtained during the preparation of Mamajjaka tablet


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Mamajjaka decoction, semisolid aqueous extract (Ghana), and tablet were analyzed by the parameters described below.

Organoleptic evaluation[10]

Organoleptic characteristics of raw herbs were shown in [Table 6].
Table 6: Organoleptic characteristics of raw herbs


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Physicochemical evaluation[11],[12]

Evaluation of physicochemical parameters mentioned in the Ayurvedic Pharmacopeia of India was carried out for raw herbs, decoction, and semisolid aqueous extract (Ghana) as shown in [Table 7].
Table 7: Physicochemical parameters of raw herbs and Mamajjaka Ghana


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Preliminary qualitative tests for functional groups[13],[14]

Mamajjaka Ghana tablet was extracted with distilled water by maceration process. The extracted samples were used for various qualitative tests to detect the presence of functional groups, which plays a very important role in the expression of biological activity as shown in [Table 8].
Table 8: Various functional groups of Mamajjaka Ghana


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Quantitative analysis

Assay of total bitter residue[15] and total alkaloid content[16] was performed by gravimetric method whereas the assay of piperine was performed by ultraviolet spectrophotometric method [Table 9].[17]
Table 9: Physicochemical parameters of Mamajjaka Ghana tablet


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  Results and Discussion Top


Mamajjaka (Enicostemma littorale blume), a perennial herb of the family Gentianaceae, is cosmopolitan in occurrence in India. It is an erect, perennial herb, 5-30 cm tall, and simple or branched at the base. Leaves are sessile and sometimes narrowed into a petiole-like base, longer than the internodes. It is of inflorescence in many flowered auxiliary clusters and numerous in the axils of each pair of leaves. Flowers are white with green lines, drying yellowish, sessile or subsessile.[18]

Daily uptake of 2 g of Enicostemma littorale fresh leaves is recommended for diabetic patients since it is highly nutritious.[19] The nutritional analysis report of the Indian Council of Medical Research suggests that 100 g of fresh Enicostemma littorale contains 140 kcal energy, 7 g of protein, 0.7 g of fat, 26.5 g of carbohydrate, 4.2 g of fiber, 8.4 g of minerals, 49.9 mg of iron, 1.641 mg of calcium, and 81 mg of phosphorous.[20]

All the clean and dried ingredients were taken. Mamajjaka whole plant was converted into coarse powder with the help of a disintegrator (pass through sieve no. 44). Other ingredients (Mamajjaka, Katuki, Ativisha, and Pippali) were converted into fine powder (powder pass through sieve no. 80). The coarse powder of Mamajjaka was subjected to overnight soaking in RO water for eight times, and the next day, it has been taken for the preparation of decoction. During the preparation of decoction, initially, the liquid was light brown and bitter. Evaporation was started at 70°C, which was aggravated on stirring. During boiling, typical irritating smell which causes nausea and burning sensation in eyes was observed. The liquid was light brown in the initial stage, which was gradually turned to greenish brown and finally into dark brown. The maximum temperature of the liquid during boiling stage was found to be between 90°C and 95°C. An average of 38.25 h were required for the preparation of Mamajjaka decoction. The average temperature maintained during the process was 95-100°C. The average yield of decoction was 168.75 L in all the four batches.

The degree of size reduction depends on the structure of the drug. Particle size reduction facilitates adequate mass transfer for better extraction. To facilitate further extraction of water-soluble constituents, overnight soaking was done, which allows micelles to take up liquid film and tissue swelling. Swelling also results from distension and bursting of thin-walled cells that have taken up the liquid by osmosis. During processing, the application of mild heat is required with occasional stirring to avoid destruction of the components sensitive to higher temperature.[21] It has been practically observed that eight times water taken for the preparation of decoction was found sufficient to extract the maximum active constituent. Hence, eight times water has been taken for the same and reduction was made up to 1/4th of its original volume. Decoction of Mamajjaka was processed further to prepare Ghana (semisolid extract). After 4 h of boiling, green-colored froth was observed which was subsided after stirring. After 12 h of heating, stickiness of the liquid and adhesiveness to the vessels were increased. Thickness of liquid and adhesiveness of vessel were gradually increased. After completion of the process, dark brown-colored semisolid extract was obtained. An average of 11.67 kg extract was obtained. Totally 46.68 kg of the extract was obtained in all the four batches. After mixing the fine powder into the semisolid extract of Mamajjaka, dark brown-colored mass was found.

Tanna et al. reported that for the preparation of Mamajjaka aqueous extract (Ghana), 2 kg coarse powder (yavkut) was taken and reduction was made up to 1/8th of its original volume. Maximum temperature given for the preparation of decoction (Kwatha) and aqueous extract (Ghana) was between 90-950 C and 70-750 C, respectively, while temperature during oven drying of the extract (Ghana) was between 45°C and 500 C. The average time required for the preparation of decoction and aqueous extract (Ghana) was 9.06 h and 7.56 h, respectively. The average yield of decoction and aqueous extract was 4.15 L and 295.6 g (14.78%), respectively.[21]

During mixing of Mamajjaka aqueous extract with Prakshepa Dravya (fine powder of Mamajjaka, Katuki, Ativisha, and Pippali, passed through sieve no. 80) and gum acacia in end runner, stickiness was observed in mass after 45 min. An average of 55 min was required for mixing of aqueous extract and fine powder of Prakshepa dravya. During drying in tray dryer after 1 day (each day 8 h), stickiness was observed in lumps and the upper layer of the lumps was dried, but the inner material was not dried properly. After 2 days of drying (each day 8 h) in tray dryer, the lumps were not dried completely, some stickiness was observed in lumps and when lumps broken, the inner material stick to the hand. When the dryer was cooled down, the lumps became somewhat hard compared to the earlier one. During preparation of granules through multi mill, pore of the multi mill blocks due to adhesive properties of extract of Mammajaka. After granulation, the granules were also dried in sunlight from 9 am to 5 pm. Granules were dried properly in sunlight. Totally 16 h were required for drying of lumps in the oven and 8 h were required for the sun drying of the prepared granules. Color of the dried granules was light brown, and some stickiness was observed in the dried granules due to the presence of Ghana (solidified aqueous extract).

Starch and talcum powder used as lubricants for the preparation of tablet tend to equalize the pressure distribution in a compressed tablet. They form a coat around the individual particles, which remains more or less intact during compression. The presence of a lubricant coating may cause an increase in the disintegration time and may also cause a decrease in drug dissolution rate. Mamajjaka Ghana is very sticky in nature, so to convert it into granule form, 960 g of pharmaceutical grade starch powder and 120 g talcum powder were added to the granules. Granulation depends on the physical characteristics of the material. After conversion of the powders into granules, their cohesiveness, compressibility, and flow properties are increased. The dissolution rate of hydrophobic drugs may be improved. Here, granules were prepared through sieve number 12. After mixing of the excipients, granules appeared light brown compared to the earlier one. Very less stickiness was observed in granules before compression. After addition of excipients (starch, talcum powder, and aerosil), free-flowing property of the granules was increased. Before starting the compression, hopper, die, punches, and all the accessories of the tablet machine were cleaned properly. After starting the operation, in-process quality control measures such as weight variation, hardness, and friability were checked out and then finally compression was carried out. Finally prepared Mamajjaka Ghana tablet was light brown. Four batches of Mamajjaka Ghana tablet were prepared to maintain the standard manufacturing procedure and the same process was repeated for the preparation of decoction, aqueous extract, and tablet in all the four batches.

All the samples of raw herbs, aqueous extract, and tablet were subjected to organoleptic and physicochemical analyses. Mamajjaka powder was greenish brown, soft and slimy, and bitter. Foreign matter, total ash, acid-insoluble ash, loss on drying, water-soluble extractive, and alcohol-soluble extractive values of Mamajjaka powder were 0.77%, 11.45%, 0.64%, 0.256%, 26.56%, 24.12%, respectively. Decoction of Mamajjaka was brown and sticky. Average pH, specific gravity, and total solid content of Mamajjaka decoction were 4.76, 1.0063, and 6.39%, respectively.

Average pH value, loss on drying, ash value, acid-insoluble ash, water-soluble extract, and alcohol soluble extract of Mamajjaka Ghana were 4.37, 4.20%, 11.29%, 0.59%, 14.1%, and 12.3%, respectively. Qualitative tests performed for various functional groups reveal the presence of alkaloids, glycosides, saponin, flavonoids, proteins, and carbohydrates in Mamajjaka Ghana.

In Mamajjaka Ghana tablet, loss on drying, water-soluble extractive, alcohol-soluble extractive, total ash, and acid-insoluble ash were 6.36%, 61.21%, 50.87%, 12.98%, and 2.46% w/w, respectively. Higher yield of extract (both water and alcohol), total ash, and acid-insoluble ash in tablet is observed due to a combination of all the ingredients. Friability, hardness, and disintegration time were 0.2%, 3 kg/cm2 , and 25 min, respectively.

Bitterness stimulates secretions in the gastrointestinal tract, especially of gastric juice. The bitter properties of herbal materials are determined by comparing the threshold bitter concentration of an extract of the materials with that of a dilute solution of quinine hydrochloride R. The bitterness value is expressed in units equivalent to the bitterness of a solution containing 1 g of quinine hydrochloride R in 2000 ml.

Total bitter residue in Mamajjaka Ghana tablet was found to be 6.13% w/w. Total alkaloid content and piperine were found to be 0.69% w/w and 0.99% w/w, respectively.


  Conclusion Top


Mamajjaka Ghana Vati is a widely used traditional formulation in the present era for its antidiabetic and antipyretic attributes. An average of 97.93% Mamajjaka Ghana tablet was obtained. The pharmaceutical and physicochemical data pertaining to large-scale production of Mamajjaka Ghana tablet are essential to assure the quality of finished products and will be useful for ayurvedic clinicians and pharmaceutical industry populace to maintain the uniformity of dosage and to maintain the batch-to-batch variations and in-process quality control measures during various pharmaceutical operations during manufacturing.

Financial support and sponsorship

This study was financially supported by the Department of Pharmacy, Gujarat Ayurved University, Jamnagar, Gujarat, India.

Conflicts of interest

There are no conflicts of interest.

 
  References Top

1.
Shrivastava S. Sharangdhar Samhita. II nd ed. Varanasi: Chaukhamba Orientalia; 1998. p. 127.  Back to cited text no. 1
    
2.
Shrivastava S. Sharangdhar Samhita. II nd ed. Varanasi: Chaukhamba Orientalia; 1998. p. 135.  Back to cited text no. 2
    
3.
Shrivastava S. Sharangdhar Samhita. II nd ed. Varanasi: Chaukhamba Orientalia; 1998. p. 208.  Back to cited text no. 3
    
4.
The Wealth of India Raw Materials Series. New Delhi: NISCAIR- CSIR; 2006. p. 174.  Back to cited text no. 4
    
5.
Sharma PV. Shodahala Nighantu Namasangraha and Gunasangraha. I st ed. Baroda: Oriental Institute; 1978. p. 74.  Back to cited text no. 5
    
6.
Billore KV. Database on Medicinal Plants Used in Ayurveda. Delhi: CCRAS; 2005. p. 312.  Back to cited text no. 6
    
7.
Bapalala G. Vaidya Nighantu Adarsha. Varanasi: Chaukhambha Bharti Academy; 2009. p. 74.  Back to cited text no. 7
    
8.
Anonymous. Bhaishajya Samhita Gujarat Rajya Bhaishajya Samiti. Ahmedabad: Health Ministry Gujarat Publisher; 1960. p. 493.  Back to cited text no. 8
    
9.
Ayurmedline. Ayurvedic Drug Index. 10 th ed. Bangalore: Dr. Seetharam Prasad Publisher; 2009. p. 469.  Back to cited text no. 9
    
10.
Kokate CK, Purohit AP, Gokhale SB. Pharmacognosy. 42 nd ed. Pune: Nirali Prakashan; 2008. p. 63.  Back to cited text no. 10
    
11.
The Ayurvedic Pharmacopoeia of India, Government of India, Ministry of Health and Family Welfare, Department of Indian System of Medicine and Homoeopathy. I st ed., Part II. Vol. 2. New Delhi; 2008. p. 15-7.  Back to cited text no. 11
    
12.
The Ayurvedic Pharmacopoeia of India, Government of India, Ministry of Health and Family Welfare, Department of Indian System of Medicine and Homoeopathy. I st ed., Part II, Vol. 1. New Delhi; 2007. p. 243.  Back to cited text no. 12
    
13.
Baxi AJ, Shukla VJ, Bhatt UB. Methods of Qualitative Testing of Some Ayurvedic Formulations. Jamnagar: Gujarat Ayurved University; 2001. p. 512.  Back to cited text no. 13
    
14.
Khandelwal KR, Sethi V, editors. Practical Pharmacognosy Techniques & Experimental. II nd ed. Delhi: Nirali Prakashan; 2001. p. 149-56.  Back to cited text no. 14
    
15.
Quality Control Method for Herbal Material, WHO. Updated Edition of Quality Control Methods for Medicinal Plant Materials. Geneva: WHO Press; 1998. p. 41-2.  Back to cited text no. 15
    
16.
Wagner H, Bladt S. Plant Drug Analysis: A Thin Layer Chromatography Atlas. II nd ed. Germany: Springer Science and Business Media; 1996. p. 22.  Back to cited text no. 16
    
17.
Vishvnath G, Jain UK. Estimation of piperine by UV-Spectrophotometric method in herbal formulation, pippli churna. Int J Res Pharm Biomed Sci 2011;2:550-3.  Back to cited text no. 17
    
18.
Gupta RS, Singh D. Hepatomodulatory role of Enicostemma littorale Blume against oxidative stress induced liver injury in rats. Afr J Agric Res 2007;2:131-8.  Back to cited text no. 18
    
19.
Upadhyay UM, Goyal RK. Efficacy of Enicostemma littorale in type 2 diabetic patients. Phytother Res 2004;18:233-5.  Back to cited text no. 19
    
20.
Sathish Kumar R, Lakshmi PT, Annamalai A. Effect of drying treatment on the content of antioxidants in Enicostemma littorale Blume. Res J Med Plant 2009;3:93-101.  Back to cited text no. 20
    
21.
Tanna SP, Patgiri B, Shukla VJ, Prajapati PK. Pharmaceutical standardization of Mamajjaka (Enicostemma littorale Auct. non Bl) Ghana. Ayu 2012;33:294-8.  Back to cited text no. 21
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