NDLI: Bioanalytical method validation of rapamycin in ocular matrix by QTRAP LC-MS/MS: application to rabbit anterior tissue distribution by topical administration of rapamycin nanomicellar formulation.

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Bioanalytical method validation of rapamycin in ocular matrix by QTRAP LC-MS/MS: application to rabbit anterior tissue distribution by topical administration of rapamycin nanomicellar formulation.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Earla, Ravinder Cholkar, Kishore Gunda, Sriram Earla, Rajya Lakshmi Mitra, Ashim K.
Description	Country affiliation: United States Author Affiliation: Earla R ( UMKC School of Pharmacy, Pharmaceutical Sciences, University of Missouri-Kansas City, 2464 Charlotte Street, 5258 Health Sciences Building, MO 64108, United States.)
Abstract	A novel, fast and sensitive 3200 QTRAP LC-MS/MS method was validated for rapamycin analysis in the rabbit eye following 0.2% administration of nanomicellar eye drop formulation. The LC-MS/MS technique was developed with electrospray ionization (ESI) in positive mode. Rapamycin was extracted from individual eye tissues and fluids by a simple protein precipitation method. Samples were reconstituted in 200µL of 80% of acetonitrile in water containing 0.05% formic acid. Twenty microliter of the sample was injected on LC-MS/MS. Chromatographic separations was achieved on reversed phase C 8 Xterra column, 50mm×4.6mm, 5µm. Multiple reactions monitoring (MRM) transition m/z 936.6/409.3 for rapamycin and 734.4/576.5 for erythromycin were employed as internal standard. The calibration curves were linear r(2)>0.9998 over the concentration range from 2.3ng/mL to 1000.0ng/mL. Rapamycin was found to be stable in ocular tissue homogenates for 6weeks at a refrigerated -80°C and -20°C temperatures. Rapamycin concentration was found to be 2260.7±507.1 (mean±S.D.)ng/g tissue and 585.5±80.1 (mean±S.D.)ng/g tissue in the cornea and iris ciliary muscle, respectively. This method has two advantages. First, a volatile base was used in the extraction procedure, which is easy to evaporate and generate consistent results. Second, the sodium adduct is employed that was stable in non-ammoniated mobile phase. The method demonstrates that absorption of rapamycin by a topical application of 0.2% rapamycin nanomicellar formulation generates therapeutically effective concentrations in the anterior segment of the eye.
File Format	HTM / HTML
ISSN	15700232
e-ISSN	1873376X
DOI	10.1016/j.jchromb.2012.09.014
Journal	Journal of Chromatography B
Volume Number	908
Language	English
Publisher	Elsevier
Publisher Date	2012-11-01
Publisher Place	Netherlands
Access Restriction	Open
Subject Keyword	Discipline Analytical Chemistry Anterior Eye Segment Chemistry Chromatography, High Pressure Liquid Sirolimus Tandem Mass Spectrometry Administration, Ophthalmic Animals Drug Stability Linear Models Micelles Nanoparticles Rabbits Reproducibility Of Results Administration & Dosage Research Support, N.i.h., Extramural Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Cell Biology Medicine Analytical Chemistry Clinical Biochemistry Biochemistry

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Bioanalytical method validation of rapamycin in ocular matrix by QTRAP LC–MS/MS: Application to rabbit anterior tissue distribution by topical administration of rapamycin nanomicellar formulation

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Bioanalytical method validation of rapamycin in ocular matrix by QTRAP LC-MS/MS: application to rabbit anterior tissue distribution by topical administration of rapamycin nanomicellar formulation.

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