The Extraction and Fermentation Process of Gallic Acid from Composition of Terminalia Species Leaves

Modelling Equations and Kinetic Parameters of Gallic Acid

Research Paper (postgraduate) 2018 54 Pages

Biology - Botany




Litterature Review

Material and Methods

Experimental Procedure






We greatly indebted to Meerut Institute of Engineering and Technology (AKTUK) that has provided me a healthy environment to drive me to achieve my ambition and goal. This institute located in Meerut, Uttar Pradesh State, India. The college is affiliated to Dr. A.P.J. Abdul Kalam Technical University, Lucknow. We convey my deep and sincere thanks to Professor Ajay Kumar Sharma (Head of the Department of Biotechnology) for extending their support and cooperation in providing access to facilities of the Department.

(D V Surya Prakash)


Gallic acid is showed best results from fermentation process than extraction process due to biotransformation. In this research, the highest concentration of gallic acid was observed to be 8.7 to 9.0µg/ml from the soxhlet extraction and column chromatography process, and the concentration was increased to 9.7µg/ml from the fermentation process by Bacillus cereus species. The fermented extracts are showed with the maximum specific growth rate and the maximum yield factor (Yx/s) of Bacillus cereus are 0.3541 hr-1 and 0.234µg/ml. The non fermented extracts are showed the modelling equation for the quercetin yield extract was Es = 0.0721(1 - e -0.2867t). The model allowed good accordance with the experimental data by producing average absolute relative deviation from about 9.78%.

Keywords: Terminalia species, Gallic acid, Fermentation, Extraction, Bacillus cereus


Terminalia arjuna

It is commonly known as Arjun tree and belongs to Combretaceae family. It is a widely used medicinal plant [15].

Origin: It is about 15 to 25 meters and growing on river banks in Uttar Pradesh, Madhya Pradesh, West Bengal and South & Central India. Stem bark is reddish colour, soft and thick. Leaves are oblong and 5-6 inch long. Flowers are yellow and white in colour [16].

Phytoconstituents : Beta sitisterol, gallic acid, ellagic acid, arjunetin, arjunic acid, ellagic acid, flavonoids like kaempferol, quercetin, phenolic acids, methyl oleanolate, terminoic acid and myristyl oleate etc.

Pharmacological activities : Cardiovascular activity, anti-inflammatory activity, anti-tumor activity, gastric activity, hepatoprotective activity, wound healing activity, antimicrobial activity, anti-oxidant activity, Immunomodulatory activity and anti-nociceptive activities etc.

Uses : It is traditionally used as a mil decoction, treatment of wounds, ulcers, fever, cough and heart diseases etc.

Scientific classification

Kingdom : Plantae

Class : Magnoliophyta

Order : Myrtales

Family : Combretaceae

Genus : Terminalia

Species : arjuna

Terminalia bellirica

It is also known as bastard myrobalan or beliric tree. It belongs to combretaceae family and large deciduous tree [18].

Origin : It is about 15-18 cm long and growing in hill regions of southeast asia, 10-15 mm thick of bark. Leaves are simple and opposite, sympodial branches, ellipsoidal seeds and flowers are bisexual and greenish yellow colour [19].

Phytoconstituents : Bellericanin, ellagic acid, chebulaginic acid, gallo-tannic acid, lignans, mannitol and phenylemblin etc.

Pharmacological activities : Anti diarrhoeal activity, Analgesic activity, Anti-microbial activity, Anti-hypertensive activity, Anti salmonella activity, Anti oxidant activity and Hepatoprotective activity etc.

Uses : It used to control the blood pressure and cholesterol, promotes hair growth and helpful to stop bleeding. The seeds contain high percentage of oil and used as biofuel. It is also used as pickle and spice.

Scientific classification

Kingdom : Plantae

Class : Magnoliophsida

Order : Myrtales

Family : Combretaceae

Genus : Terminalia

Species : bellirica

Terminalia chebula

It is also known as Chebulic myrobalan and Black myrobalan. It belongs to combretaceae family and highly used in homeopathic and ayurveda medicine. It is a moderate tree and used for the treatment of number of diseases [1].

Origin : It is deciduous and grows up to 32 m in India, China, Sri Lanka, Bangladesh, Thailand and Malaysia. The leaves are oval and 6-7 cm in long. Fruits are orange, yellow and brown in colour.

Phytochemical constituents : chebulinic acid, Chebulic acid, chebulagic acid, gallic acid, ellagic acid, flavonoids (quercetin, rutin and kaempferol), chebulosides, tannic acid, tannins, luteic acid and other constituents [4].

Pharmacological activities : Antioxidant activity, Immunomodulatory activity, Anti caries agent,

Anti‐aging activities, Anti lithiatic activity, Hypo lipidemic activity, Antidiabetic activity, Hepato‐protective activity, Anti ulcerogenic, Anti-cancer and Anti-microbial activities etc.

Uses : Chebula powder is control the various wounds, improves digestion and this herb is used as tooth paste and tonic for stronger teeth and cough. It is control the cholesterol in the body

Scientific classification

Kingdom : Plantae

Class : Magnoliophsida

Order : Myrtales

Family : Combretaceae

Genus : Terminalia

Species : chebula

Gallic acid

It is a phenolic acid and trihydroxybenzoic acid found in the plants. It is in white and yellow color crystals. It is commonly used in the pharmaceutical industry [2].

Abbildung in dieser Leseprobe nicht enthalten

Chrmical formula : C7H6O5

Density : 1.694 g/cm3

Solubility : soluble in acetone, ether, glycerol and alcohol.

Uses : Its supplements are available as ointments and capsules to treat a variety of wounds. Its control the blood pressure.

Pharmacological activities: Anti-oxidant activity, Anti-inflammatory effect, Anti - cancer effect, Anti-diabetic effect and Anti-fungal activity etc.

Assay method : Folin-Ciocalteau assay


Chattopadhyay and Bhattacharrya, 2007 studied the Terminalia chebula (Combretaceae) is being used for the treatment of various types of diseases, because of its strong power of healing with a wide spectrum of biological activity. Terminalia chebula chemical constituents, has been achieved regarding the biological activity and medicinal applications of the herbal plant. It is a development of medicines and industrial products against diseases. It possess anti-oxidant activity, anti-diabetic activity, anti-inflammatory activity, anti-microbial, anti-pyretic, anti-tumor and anti-mutagenic activities etc. The bioactive components gallic acid, ellagic acid, tannins, chebulinic acid, chebulagic acid, bellericanin, rutin, quercetin etc are presence in the herbal plant.

Patel Madhavi et al., 2010 studied that Triphala is a herbal composition containing Terminalia chebula, Phyllanthus emblica and Terminalia belerica fruits in equal proportions. The preparation is frequently used in ayurvedic medicine to treat diseases. A simple Reverse phase - High Pressure Liquid Chromatography (RP-HPLC) method for the separation and quantitative determination of phenolic compounds like gallic acid, ellagic acid and chebulinic acid components from Triphala herbal composition. In this analysis method, the use of an RP18 column with acidic mobile phase allowed the separation of compounds gallic acid, ellagic acid and chebulinic acid compounds. It takes the time period upto 30 min. The proposed Reverse phase - High Pressure Liquid Chromatography method was found to be simple, precise and accurate and can be used for the quality control of the formulations.

Anil Mahajan and Nandini Pai, 2011 explained on Triphala is one of the mainly popular herbal preparations in the world used to prepared as drugs. It is a combination of Terminalia chebula, Phyllanthus embilica and Terminalia bellirica species. A simple, precise and accurate high pressure liquid chromatography method has been developed for the determination of chebulinic acid, gallic acid, ethyl gallate, methyl gallate, chebulagic acid, ellagic acid, penta-O-galloyl-ß-D-glucose and 2,4-chebulyl-ß-D-glucopyranose in triphala. The analysis method was attained on Inertsil ODS 3 column (100mm×4. 6mm, 3µm). A mixture of acetonitrile and 0.02% triethyl amine aqueous pH 3.0 was used as mobile phase in incline mode at a flow rate of 1ml/min at 272nm. This method was carried out for linearity, precision, limit of detection and quantification. The advantage of the method is simple extraction process and short run time intervals at 27min.

Ashwini et al., 2011 informed that Terminalia chebula is a moderate herb used in traditional medicines. Terminalia chebula showed that it possess antimicrobial, antitumor, antidiabetic, antioxidant, antiulcer and wound healing activity. It has also been found that it prevents the kidney damage and also used for the treatment of heart disease. It is also an active ingredient of a herbal composition called as Triphala which comprises of three medicinal herbs namely Phyllanthus embilica (Amla), Terminalia bellerica and Terminalia chebula (Haritaki) species. This Triphala is plays a key role for the treatment of various diseases in the World. It contains phytochemical components like gallic acid, ellagic acid, tannins, chebulinic acid, chebulagic acid, bellericanin, rutin, quercetin etc. Terminalia chebula is a natural origin and it have no side effects in the medical field.

Gowda et al., 2012 reviewed on Triphala is one of the most powerful herbal drug used in medicine. Triphala is a combination of Terminalia chebula, Phyllanthus emblica, and Terminalia belerica species. It is also in the treatment of Acquired immune deficiency syndrome and various diseases. It contains many polyphenolic components, gallic acid, ascorbic acid (Vitamin C), ellagic acid, chebulinic acid, chebulagic acid, and flavonoids (quercetin and rutin). It shows the different pharmacological activities like anti-inflammatory activity, anti-cancer activity, anti-oxidant activity, anti-ulcer activity, anti-microbial activities etc.

Anwesa Bag et al., 2013 explained the medicinal plant Terminalia chebula (Haritaki) is extraordinary strong power of healing plant and have more medicinal value and traditionally used for the treatment of various diseases. People used this plant in the treatment of different ailment like vomiting, ulcers, bladder diseases, throat, asthma etc. It shows various pharmacological and medicinal activities, such as anti-inflammatory, anti-microbial, anti-oxidant, anti-arthritic, anti-diabetic, anti-tumor, anti-caries, and wound healing activity. The phytochemical constituents are isolated and identified by using the high pressure liquid chromatography (HPLC) and R-HPLC. It contains high phenolic compounds.

Ranjeet Sawant et al., 2013 reported that the Terminalia chebula, which is a belongs to the Combretaceae family, is frequently used as medicinal herb to treat most of the systemic disorders. Phyto-chemical constituents are isolated from Terminalia chebula has potential used in medical field. The phytochemical constituents are isolated and recognized by using the HPLC (high pressure liquid chromatography) and R-HPLC (reverse phase high pressure liquid chromatography). In Ayurvedic system this one commonly used as drug medicinal herbal plant for treatment of various diseases of humanbeings. Various pharmacological activities of this plant gives a good results to the medical system.

Kamali Seyed Hamid et al., 2013 reported that Triphala is one of the oldest rasayana drug and used polyherbal formulation in the medical system. It is a composition of medicinal herbs and comprised of Terminalia chebula, Terminalia bellerica and Phyllanthus embilica. This herbal combination have healing benefits in multi-organ systems and treated various diseases. And, it exhibits a number of health benefits like antioxidant activity, anti-diabetic, anti-inflammatory, anti-obesity, antigastric ulcers, and anti-mutagenic activity, chemo-preventive potential, wound healing, antimicrobial, radioprotective effect, anti-hypercholesterolemic, enteroprotective efficacies, nitric oxide scavenging activity and immunomodulatory. It is rich in megnesium, potassium, calcium, ferrous and zinc, which enhance their bioavailability. It plays a therapeutic agents and clinically used as drugs for treatment of diseases.

Rathinamoorthy and Thilagavathi, 2014 explained the Terminalia chebula is a medicinal plant widely distributed in the World and belongs to the family Combretaeae. This plant have been used to treat different diseases and infections in traditional medicinal systems and it is called the King of Medicine. The pharmacological studies on Terminalia chebula species, this gives a wide knowledge about the herb and showed various activities.

Amit Kumar, 2014 states that Ayurveda is the oldest medical system. In the last some years the field of ayurveda herbal medicine and drugs are gaining positions in various countries. The medicinal herbs are shows natural origin and without side effects. The WHO has selected up to twenty one thousand herbal plants, which are used for medicinal purposes around the world. Mostly number of professional scientists discovered that one of the most value of all Ayurvedic combination - Triphala (Terminalia chebula, Phyllanthus embilica and Terminalia bellirica) in different ratios exhibits a number of health benefits, including various pharmacological activites like antioxidant, anticancer, antidiabetic etc.

Rukshana Irani and Kazi layla Khaled , 2015 given a reviewed that Innate power of the traditional medicine is Triphala. It is one of the most significant and efficient polyherbal formula acknowledged in ayurveda is Triphala for the prevention and treatment of various health ailments. It is prepared by mixing the powdered fruits of Emblica officinalis (Amalaki), Terminalia chebula (Haritaki) and Terminalia bellerica (Bibhitaki or Bahera) in equal proportion. It is credited with diverse beneficial properties and has been reported to possess anti-oxidant activity, anti-diabetic activity, anti-inflammatory activity, anti-pyretic, anti-microbial activity, anti-tumor activity, anti-mutagenic activity, immunomodulatory, antistress, anti-cancer, antigout, anti-ulcer, wound healing, radioprotective, hepatoprotective, chemopreventive and anticariogenic effects. The bioactive components phyllembic acid, ascorbic acid, carotene, gallic acid, ellagic acid, tannins, chebulinic acid, chebulagic acid, bellericanin, rutin, quercetin etc.

Shaeesta Khaleelahmed Bhavikatti et al., 2015 given the reports that Triphala is an important medicinal herbal drug, used for treatment of various diseases. The three fruits (Phyllanthus emblica, Terminalia belerica, and Terminalia chebula) process the tonifying, antiseptic detoxifying, and laxative effects of this combination. In Ayurveda, Triphala is being widely studied and researched in the anti‑tumor, antioxidant, anti‑inflammatory, anti‑bacterial and anti-viral activities. According to the survey reports, many patients are recoverd by these Triphala drugs and gives the best results in the medical field. It contains bio compounds (phenolic compounds, tannins, phenolic acids, minerals) and associated into the various pharmacological activites.

Neethu S Kumar et al., 2016 explained the Triphala is one of the most valuable herbal preparations in the world. Triphala is an ayurvedic herbal preparation and composed of three equal proportions of herbal plants - Terminalia chebula (Haritaki), Phyllanthus emblica (Amla), and Terminalia belerica species. Triphala is rich amount of polyphenols, possess antimicrobial, antiinflammatory, antidiabetic and antitumor activities. It is also in the treatment of AIDS. It contains polyphenols, tannins and flavonoids. It is a polyherbal composition and the mechanism of action of drugs. It contains a number of phytocompounds, including gallic acid, ellagic acid, chebulagic acid, chebulinic acid etc. Triphala has a play role on nutritional as well as liver and blood cleansing activities. The dietary nutritional values of triphala in the form of its ascorbic acid (vitamin C), presence of linoleic oil and other important nutrients make it more of a tonic. Triphala possess free radical scavenging, anti-oxidant, anti-inflammatory, anti-pyretic, analgesic, antimicrobial, anti-mutagenic, wound healing, anti-cariogenic, anti-diabetic activities.

Gull Mohd Mir and Bilal Ahmad Dar, 2016 says that Triphala a rasayana drug and polyherbal formulation used in Indian System of Medicine (ISM) for various diseases. It is a composition of Emblica officinalis (Euphorbiaceae), Terminalia bellerica Linn (Combertaceae) and Terminalia chebula (Combertaceae) with 1:1:1 ratio. It is one of the best ayurvedic herbal combinations. Triphala is tridoshic rasayana in ayurvedic system. According to literature survev fifty thousand people are dying every day due to pneumonia, diarrhoea, tuberculosis and malaria diseases. Triphala is plays a key role for controlling these diseases. It contains rich phytoconstituents and possess various pharmacological activities.

Padmaa Paarakh , 2010 says that Terminalia arjuna (Roxb.) Wt. and Arn. (Arjuna; Combretaceae) is a widely used medicinal plant throughout India and popular in various Indigenous System of Medicine like Ayurveda, Siddha and Unani. In the Indian System of Medicine, the bark are used as astringent, cooling, tonic, aphrodisiac, cardiotonic, in fractures, ulcers, spermatorrhoea, leucorrhoea, diabetes, cough, tumour, excessive perspiration, asthma, inflammation and skin disorders etc.

Bariya Aditi Hemrajbhai et al., 2016 Medicinal plants have been a major source of therapeutic agents to cure human diseases, since ancient time. Terminalia arjuna is one kind of widely used medicinal plant used in various indigenous system of medicine like Ayurveda, Sidda and Unani. This review has been conducted to pile up phytochemical as well as pharmacological information that is available in different scientific literatures. This plant has been reported to contain phytochemical constituents like Triterpenoids (arjunolic acid, arjunic acid, arjunin, terminic acid, etc.), Glycosides (arjunetin, arjunosides, arjunglycosides etc.), Flavonoids (Arjunone, arjunolone etc.), Tannins (casuarinin, gallic acid, pyrocatechols etc.), β-Sitosterol, Minerals (calcium, magnesium, zinc, copper etc.). which exhibit various pharmacological activities like Antimicrobial, Anti cancer, Cardioprotective, Anti fungal, Anti diabetic, Antioxidant, Anti inflammatory, Hypolipidemic, Anthelmintic, Insecticidal, Wound healing, Anti acne, Gastroprotective etc. The present comprehensive review is therefore an effort to give detailed information on phytochemical and pharmacological studies of T. arjuna.

Aghera Hetal et al., 2015 The knowledge of Ayurveda is probably one of India’s greatest offerings to human empire. It is eternal in the profound sense that it remains pertinent for all times. There are plentiful articles found regarding Ayurvedic single drugs and formulations, however, most of them are with the deficiency of permutation of modern aspect and Ayurveda aspect. Hence, the review of Terminalia Arjuna (Roxb.) Wt. & Arn and its valuable formulation has been made with classical facts with logical portrayal, including evidences from Vedas, Samhitas, Medicinal books, Tribal colonies and different search engines on podium. This review article reflects history, properties, chemical constituents, pharmacological actions, toxicity, and different uses of plant Terminalia Arjuna (Roxb.) Wt. & Arn. and reasonable role of various formulations and media incorporate with the same plant. In sum up, this sort of review would serve as the most important step to engender new approach of Ayurvedic drug research, a path to sense possible alterations in drug formulations and ultimately new drug discovery

Anindita Deb et al., 2016 Terminalia bellerica Roxb (combretaceae) is found widely throughout the Indian subcontinent, Sri Lanka, South- East Asia, Bangladesh as a medicinal plant. Plant and plant parts are used in the traditional system of medicines like Ayurveda, Siddha, Unani & Chinese medicine. The plant is constituted of Glucoside, Tannins, Gallic acid, Ethyl Gallate, Chebulinic acid which serves as an antioxidant, antimicrobial, antidiarrheal, anticancer, antihypertensive, hepatoprotective& antipyretic agent. This review sites the information on pharmacological activities of Terminalia bellerica which may serve as a source for further research studies.

Saraswathi Motamarri et al., 2012 Terminalia belerica. Roxb (TB) is growing widely throughout the Indian subcontinent, Sri Lanka and SE Asia. In the Traditional system of medicine (TCM) like Ayurveda, Siddha and Unani, medicinal uses have been described as it is works in disease of every system. Glucoside, Tannins, Galliacid, Ellagicacid, Ethylgalate, Gallylglucose, Chebulanic acid are mainly believed to be responsible for its wide therapeutic actions. It is used as antioxidant, antimicrobial, antidiarrheal, anticancer, antidiabetic, antihypertensive and hepatoprotective agent. The present article attempts to provide comprehensive information on phytopharmacological properties of Terminalia belerica for further research.

Pedro Aguilar-Zárate et al., 2015 Tannase is an enzyme that catalyses the breakdown of ester bonds in gallotannins such as tannic acid. In recent years, the interest on bacterial tannases has increased because of its wide applications. The lactic acid bacteria (LAB) plays an important role in food tannin biotransformation, it has the ability of hydrolyse tannins in ruminants intestine. The finding of tannin hydrolysis by LAB has sparked their use as tannase producer. The bacterial strains used in the present work were identified as Bacillus subtilis AM1 and Lactobacillus plantarum CIR1. The maximal tannase production levels were 1400 and 1239 U/L after 32 and 36 h of fermentation respectively, for B. subtilis AM1 and L. plantarum CIR1. Maximum gallic acid release was 24.16 g/L for B. subtilis AM1 and 23.73 g/L for L. plantarum CIR1. HPLC analysis showed the formation of another peaks in the retention time range of 9–14 min, which could be attributed to the formation of di or tri-galloyl glucose.



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Title: The Extraction and Fermentation Process of Gallic Acid from Composition of Terminalia Species Leaves