HPTLC-densitometry quantification of fructooligosaccharides from inulin hydrolysate

The objective of present research was to develop an easy, precise and accurate HPTLC densitometry method for quantification of fructooligosaccharides (FOSs) from inulin hydrolysate. The chromatographic separation of FOSs was performed on pre-coated silica gel (60, F₂₅₄) TLC plates using a mobile phase (butanol:ethanol:water, 60:24:16), and densitometry evaluation of FOSs was performed at A₅₀₀. Both kestose and nystose were successfully resolved with R_f value of 0.43 and 0.34, respectively. The accuracy, reliability and reproducibility of developed method was assessed by percent relative standard deviation of kestose and nystose for instrument precision (1.43% and 1.50%), repeatability (1.48% and 1.56%), intra-day precision (1.60% and 1.63%), inter-day precision (1.62% and 1.66%), limit of detection (4.58 ng/spot and 4.58 ng/spot), limit of quantification (13.87 ng/spot and 13.89 ng/spot) and recovery (98.81% and 98.69%). Moreover, overlapping spectra of test sample with standard confirms the specificity of developed method, which was validated as per ICH guidelines.