Development and validation of TLC-densitometric method for determination of lipid A adjuvant as a bulk and in solid fat nanoemulsions

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 si...

Full description

Saved in:
Bibliographic Details
Published in:Biomedical chromatography 2015-10, Vol.29 (10), p.1473-1479
Main Authors: Minz, Sunita, Kaurav, Monika, Sahu, Kantrol Kumar, Mandal, Vivekananda, Pandey, Ravi Shankar
Format: Article
Language:English
Subjects:
Calibration
Chromatography, Thin Layer - instrumentation
Chromatography, Thin Layer - methods
Densitometry - methods
Emulsions - analysis
Emulsions - chemistry
high-performance thin-layer chromatography
Limit of Detection
Linear Models
Lipid A - analysis
monophosphoryl lipid A
Nanostructures - analysis
Nanostructures - chemistry
Sensitivity and Specificity
solid fat nanoemulsions
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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 Rf 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 r = 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.
ISSN:0269-3879
1099-0801
DOI:10.1002/bmc.3444