Preparation Of Anti-human IgG-hrp Conjugate And Its Use In ELISA And Western Blotting Experiments | 5891
ISSN: 2155-9872

Journal of Analytical & Bioanalytical Techniques
Open Access

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Preparation of anti-human IgG-hrp conjugate and its use in ELISA and western blotting experiments

3rd International Conference and Exhibition on Analytical & Bioanalytical Techniques

K. Ramesh Kumar

Posters: J Anal Bioanal Techniques

DOI: 10.4172/2155-9872.S1.010

A highly sensitive method for detection and quantification of swertiamarin, mangiferin and amarogentin in Swertiachirata (Gentianaceae).The leaves, roots, stem, flowers and tissue cultured plants were extracted with 80% methanol in a bath sonication for rapid extraction of metabolites. The desired separation and baseline was obtained in a gradient method where mobile phase A was composed of 0.1% TFA in water and mobile phase B was a mixture of acetonitrile: water in the ratio of 70:30. The linear gradient at a flow rate of 1.0ml/min was start with 15% B; 20% B in next 5 min, 70% B in next 25 min, hold for 5 min; 15%B in next 5 and equilibrated for 5 min at 240 nm UV wavelength. The separation was obtained on Waters Spherisorb 5μm ODS2 (250 mm x 4.6 mm). The LOD of swertiamarin, mangiferin and amarogentin were found to be 0.4μg/ml, 0.2μg/ml and 0.3μg/ml respectively. The LOQ of swertiamarin, mangiferin and amarogentinwere found to be 1.0μg/ml, 0.4μg/ml and 0.9μg/ ml respectively. The method can be used as quality control of plant material and also identification of Swertiachirata from other species of Swertia due to the presence of marker compound amarogentin.
Varun Kumar have completed his M. Pharmacy from GJU, Hissar and is having 2 years of Research experience in Analytical research and development, Jubilant Life Sciences. Now pursuing PhD in Jaypee University of Information Technology, Shimla Hills.