Nonspecific Binding in LC–MS Bioanalysis

Tan, Aimin; Fanaras, John C.

doi:10.1002/9781119413073.ch9

Cited by 5 publications

(1 citation statement)

References 40 publications

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“…For tissue homogenization, diluted rabbit plasma was chosen to provide sufficient amounts of protein to compete for and solubilize the analytes from various tissue matrices during homogenization as well as to reduce nonspecific binding (Tan & Fanaras, ). Various dilutions of rabbit plasma (5‐, 10‐, 15‐ and 20‐fold) were tested.…”

Section: Resultsmentioning

confidence: 99%

Simultaneous quantification of candesartan and irbesartan in rabbit eye tissues by liquid chromatography–tandem mass spectrometry

Tan

Gui

Wong

et al. 2020

Biomedical Chromatography

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Diabetic retinopathy is a major cause of vision loss in adults. Novel eye-drop formulations of candesartan and irbesartan are being developed for its cure or treatment. To support a preclinical trial in rabbits, it was critical to develop and validate a new LC-MS/MS method for simultaneous quantification of candesartan and irbesartan in rabbit eye tissues (cornea, aqueous humor, vitreous body and retina/choroid). Eye tissue samples were first homogenized in H 2 O-diluted rabbit plasma. The candesartan and irbesartan in the supernatants together with their respective internal standards (candesartan-d 4 and irbesartan-d 4 ) were extracted by solid-phase extraction. The extracted samples were injected onto a C 18 column for gradient separation. The MS detection was in the positive electrospray ionization mode using the multiple reaction monitoring transitions of m/z 441 ! 263, 445 ! 267, 429 ! 207, and 433 ! 211 for candesartan, candesartan-d 4 , irbesartan and irbesartan-d 4 , respectively. For the validated concentration ranges (2-2000 and 5-5000 ng/g for candesartan and irbesartan, respectively), the within-run and between-run accuracies (% bias) were within the range of −8.0-10.0. The percentage CV ranged from 0.6 to 7.3. There was no significant matrix interference nor matrix effect from different eye tissues and different rabbits. The validated method was successfully used in the Good Laboratory Practice (GLP) study of rabbits. K E Y W O R D S candesartan, eye tissue, irbesartan, LC-MS, rabbit

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Section: Resultsmentioning

confidence: 99%