Development and validation of TLC‐densitometric method for determination of lipid A adjuvant as a bulk and in solid fat nanoemulsions. (24th February 2015)

• Record Type: Journal Article
• Authors: Minz, Sunita; Kaurav, Monika; Sahu, Kantrol Kumar; Mandal, Vivekananda; Pandey, Ravi Shankar
• Is Part Of: Biomedical chromatography. Volume 29:Number 10(2015:Oct.)
• Page Start: 1473
• Page End: 1479
• Publication Date: 2015-02-24
• Subjects: Chromatographic analysis -- Periodicals; Biology -- Periodicals; Medicine -- Periodicals; Biology -- Periodicals; Chromatography -- methods -- Periodicals; Medicine -- Periodicals; 543.089
• DOI: 10.1002/bmc.3444
• Languages: English
• ISSNs: 0269-3879

Abstract

<abstract abstract-type="main"> <title>Abstract</title> <p>A simple, sensitive, selective and precise high‐performance thin‐layer chromatographic method was developed for determination of lipid A (MPLA) adjuvant as a bulk and in solid fat nanoemulsions. Chromatographic separations were performed on thin‐layer chromatography aluminum plates precoated with silica gel 60 F‐254 as stationary phase and chloroform–methanol–ethyl acetate solution (10:2:4, v/v/v) as mobile phase. With this solvent system, compact spots for MPLA at <italic>R</italic><sub>f</sub> value 0.80 ± 0.02 were obtained. Densitometric analysis of MPLA was carried out in absorbance mode at 357 nm. Linear regression analysis for the calibration plots showed good linear relationship with <italic>r</italic> = 0.9996 in the concentration range of 20–100 ng/spot. The mean values (±SD) of slope and intercept were found to be 7.355 ± 0.006 and 109.52 ± 0.170, respectively. Limits of detection (LOD) and quantitation (LOQ) were observed at 3.096 and 9.382 ng/spot, respectively.The method was validated for precision, accuracy, robustness and recovery as per the International Conference on Harmonization guidelines. Statistical analysis proved that the developed method for quantification of MPLA as a bulk and in solid fat nanoemulsions is reproducible, selective and economical. This method could be applied for quantitative assay of MPLA in lipid‐based vaccine formulations. Copyright © 2015 John Wiley & Sons, Ltd.</p> </abstract>