Quantification of Hypocrellin B in Rabbit Ocular Tissues/Plasma by Spectrofluorimeter: Application to Biodistribution Study

Author(s): Venkateshwaran Krishnaswami, Chandrasekar Ponnusamy, Senthilkumar Sankareswaran, Muthuselvam Paulsamy, Rajaguru Palanisamy, Subramanian Natesan.

Journal Name:Current Bioactive Compounds

Volume 10 , Issue 4 , 2014

Abstract:

A simple and sensitive red spectrofluorimetric method has been described for the estimation of hypocrellin B in rabbit plasma and ocular tissues such as retina, iris, aqueous humour and vitreous body. The fluorimetric estimation was achieved using black coloured 96- well plate at an excitation/emission wavelengths of 470/620 nm with a specific well volume of 100 µL/well. Hypocrellin B calibration curves were linear at a concentration ranging from 20 ng/mL to 1280 ng/mL with correlation coefficient between 0.86 and 0.99 in ocular tissues and plasma. Limit of detection and quantification was between 0.011 to 0.017 µg/mL and 0.034 to 0.054 g/mL respectively for hypocrellin B in different ocular tissues and plasma. The percentage relative standard deviation observed for the validation parameters such as method precision, system precision, intra- inter day, analyst to analyst variations and robustness is found to be less than 12.5 % for the observed fluorescence intensity in different ocular tissues/plasma. The method has been applied to check the hypocrellin B concentration in plasma and ocular tissues after intravenous administration and found that hypocrellin B has some exposure to the posterior segment of eye without major intraocular presuure abnormalities. This selective and sensitive fluorimetric method will facilitate the future investigations of hypocrellin B on ocular delivery of hypocrellin B with less solvent consumption.

Keywords: Calibration, emission, excitation, eye, fluorimetric, hypocrellin B.

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Article Details

VOLUME: 10
ISSUE: 4
Year: 2014
Page: [245 - 253]
Pages: 9
DOI: 10.2174/1573407210666141001000336
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