Development and validation of an HPLC-fluorescence method for the quantification of IR780-oleyl dye in lipid nanoparticles

Varache, Mathieu ORCID: https://orcid.org/0000-0001-7166-2253, Escudé, Marie, Laffont, Corentin, Rustique, Emilie and Couffin, Anne-Claude 2017. Development and validation of an HPLC-fluorescence method for the quantification of IR780-oleyl dye in lipid nanoparticles. International Journal of Pharmaceutics 532 (2) , pp. 779-789. 10.1016/j.ijpharm.2017.06.019

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Abstract

A reversed-phase (RP) high-performance liquid chromatography (HPLC) method for the content determination of IR780-oleyl (IRO) dye in lipid nanoparticles was developed and validated. Chromatographic separation was performed on a RP C18 column with a gradient program of water and acetonitrile both with 0.1% (v/v) TFA, at a flow rate of 1.0 mL/min and a total run of 21 min. IRO dye detection was made by fluorescence at emission wavelength of 773 nm (excitation wavelength: 744 nm). According to ICH guidelines, the developed method was shown to be specific, linear in the range 3–8 μg/mL (R2 = 0.9998), precise at the intra-day and inter-day levels as reflected by the coefficient of variation (CV ≤ 1.98%) at three different concentrations (4, 6 and 8 μg/mL) and accurate, with recovery rates between 98.2–101.6% and 99.2–100.5%. The detection and quantitation limits were 0.41 and 1.24 μg/mL, respectively. Stability studies of sample processing showed that IRO dye was stable after 24 h in the autosampler or after three freeze/thaw cycles. Combined with fluorescence measurements, the developed method was successfully applied to optimize the loading capacity of IRO dye in the core of lipid nanoparticles.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Dentistry
Subjects:	R Medicine > RK Dentistry
Uncontrolled Keywords:	Analytical method; Fluorescence detection; Lipid nanoparticles; High performance liquid chromatography; Near infrared dye
Publisher:	Elsevier
ISSN:	0378-5173
Date of First Compliant Deposit:	4 July 2017
Date of Acceptance:	7 June 2017
Last Modified:	19 Jan 2025 15:15
URI:	https://orca.cardiff.ac.uk/id/eprint/102018

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