IJCRR - Vol 09 Issue 22, November
Date of Publication: 30-Nov--0001
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Variation of Andrographolide Content in Andrographis paniculata from Different Sites of Balaghat Region of Jabalpur (M.P.)
Author: Anil Kushram, S. K. Masih, Sajad Ahmad Mir
Category: General Sciences
Abstract:Aim: The main objective of this investigation was to show the variation of Andrographolide content in Andrographis paniculata from thirty seven different sites of Balaghat (Jabalpur MP) region.
Methodology: In order to study the variation in andrographolide content, plant material was collected from different sites and were analyzed through quick and accurate HPLC chromatographic method using C-18 column (4.6 x 250 mm, 5\? particle size).
Result and Discussion: The average andrographolide content varied from 0.01 to 1.2%. Highest percent of andrographolide in Andrographis paniculata were estimated from site Kera of south Balaghat 1.2%. Lowest content of andrographolide in Andrographis paniculata were estimated from the site of Salhen north Balaghat region that is 0.02%.
Conclusion: The content of this active ingredient in this plant varies with in the plant parts and with the geographical distribution.
Keywords: Andrographis paniculata, Andrographolide, Balaghat region, HPLC instrumentation
Andrographis paniculata is an important medicinal plant of family acanthaceae. It is also called king of bitters. It is an annual herb widely used in different parts of Asia. Used mainly for treating fever, liver disease, common cold, chickengunia fever, snake bite etc. It is to be used extensively for medicinal purpose in India, Thailand, China and Mauritius (Niranjan, et. al., 2010). The leaves and aerial parts of the plant are used in Indian traditional medicine for the treatment of fever, malaria, some throat infection (Shanker, et. al., 2012), hepatoprotective, antioxidant (Lin, et. al., 2009). Whole plant has variety of therapeutic uses. Phytochemical data revealed that this plant can be used for the treatment of common cold pharyngotonsillitis and diarrhea (Mishra et al 1992 and Najib et al 1999). Antioxidant properties of Andrographis paniculata were recorded by (Trivedi and Raval, 2001). This plant is used for blood purifier, liver diseases, dermatological diseases (Prathanturarug et al., 2007). Aqueous extract of this plant are having antimicrobial activity due to presence of arabinogalactan proteins and andrographolide (Singh et al., 2003). Leaf part was found to content highest amount of andrographolide and the seeds were estimated to content lowest amount of andrographolide (Sharma et al., 1991). Andrographolide is bitter crystalline compound, colorless having antithrombotic, hyphoglycemic and antipyretic properties. A recent study conducted by USA University that this plant is having anti HIV activity (Calabrese, 2000). This plant ranks seventeenth position among their thirty two prioritized medicinal plants and annual growth of this plant is 3.1 percent (Anonymous, 2007).
MATERIALS AND METHODS
Plant material: Andrographis paniculata was collected from different sites of Balaghat region. Areal plant parts were collected for comparative study of andrographolide content. Leaves/ plant material at different sites were collected and dried in shade for 15 days followed by grinding to form powder of it. The fine powdered samples were used in methanol solvent to estimate andrographolide content by HPLC method.
HPLC Instrument: HPLC instrument that was used for the estimation of andrographolide were of the following features, HPLC- grade waters, Pump - 515 Isocratic pump, Injector - Rheodyne injector with a 20-microlitre loop, Detector - UV Vis detector, Software - Data ace software, Column - Thermo C-18 column (4.6 x 250mm, 5μ particle size), sample size (20µl). Isocratic elution was carried out with methanol at a flow rate (1ml/min). The detection was performed with wavelength (230 nm) and column temperature was ambient (300C). Class VP software was used for integration and calibration. Evaluation was via peak areas with linear regression.
Preparation of herbal extract: Fresh aerial part of the plant sample collected from 37 different sites of Balaghat region of Jabalpur (M.P). Reflux 1 gm dried powder along with 50 ml of methanol was kept in soxhlet for one hour. After one hour the refluxing load was subjected to Rota-vapor at 60 RPM and heated at 600C. Filter & subject the marc for another two cycles of refluxes (1 hrs. each) with methanol (50 ml) combine with the filtrate. Evaporate under vacuum to dryness Dissolve the residue 10 mg in methanol (10ml). Filter, Inject the solution in HPLC with the help of 20 µl fixed loop injector and percent content of andrographolide were estimated by counting the area of andrographolide peak in HPLC chromatogram in all sample.
Table 1: Variations of Andrographolide content from Andrographis paniculata
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