Organically grown Andrographis paniculata plant was obtained from Jeevaniya Society, Lucknow, India and identified by CSIR-NISCAIR Herbarium and Museum, New Delhi. The aerial parts of the plant was removed, washed with distilled water and dried in an oven at 60 °C overnight and converted into powder by pulverization. 100 gm of the powder was extracted with 1 litre of Ethanol: Water (1:1 v: v) at 40 °C in a water bath shaker overnight. The extract was filtered and the clear filtrate was evaporated in a rotary evaporator at reduced pressure to 100 ml of slurry. 20 ml of this slurry was loaded on to a silica gel (Merck, India, 60–120 mesh) column (20 cm × 2.5 cm) using hexane and eluted sequentially with 200 ml each of hexane followed by analytical-grade ethyl acetate, acetone and finally with methanol. Each solvent fraction was evaporated and the residual matter was examined by TLC by comparing with authentic andrographolide (Sigma Aldrich-USA). The material present in the acetone fraction was crystallized from methanol to give yellow crystals which was identified to be andrographolide on the basis of its thin layer chromatography (TLC) profile, melting point, and High Performance Liquid Chromatography (HPLC) and 1H NMR profiles and compared with authentic andrographolide (Sigma Aldrich-USA).
Authentic andrographolide
Manufactured by Merck Group
Sourced in United States
Authentic andrographolide is a purified compound derived from the herb Andrographis paniculata. It is a key active ingredient used in various research and laboratory applications. The compound has well-documented chemical and physical properties that make it a valuable tool for scientific investigations.
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Lab products found in correlation
2 protocols using authentic andrographolide
1
Extraction and Characterization of Andrographolide
2
Andrographolide Quantification from Plant Tissue
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About 50 mg of dried plant tissue powder from different plant parts was subjected to methanol extraction as per Zaheer and Giri [37 ] with minor modifications. Dried powder was added to 5 m of HPLC grade methanol (Fisher Scientifics, India) and incubated at room temperature for 24 h followed by 1 h sonication in Ultrasonic Cleaning Bath (Spectralabs Instruments Pvt. Ltd, Mumbai, India). The methanol extract was filtered through Whatman No.41 filter paper first and then through 0.2 µm Millex GV Durapore PVDF filter (Merck, Germany). The filtered sample (500 µl) was used for analysis in the Waters HPLC system (Waters, USA) using a C18 column. The methanol (100%) was used as an isocratic solvent system. The samples were run for 15 min with 20 µl injection volume and 1 mL/min flow rate at 25 °C. The pressure was maintained in the range of 2000–6000 psi. The andrographolide content was detected with the PDA detector. The authentic andrographolide (98%) procured from Sigma-Aldrich (USA) was used as a standard. Andrographolide content was estimated by calculating peak areas and comparing with the authentic standard.
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