Optimization and validation of UV-VIS spectrophotometry methods on the standardization of galactomannan levels inguar gum (Cyamopsis tetragonoloba L.)

Abstract

Aim: Method validation is a step to ensure that the method used is in accordance with the desired goals. The Category
I validation method determines the content of the main ingredient in the drug substance. Galactomannan is a watersoluble
polysaccharide composed of D-galactose and D-mannose and is the main component in guar gum. Materials
and Methods: To determine galactomannan content, concentrated phenol-sulfuric acid was used using the UV-Vis
Spectrophotometry method. Hydrolysis of guar gum is required to react with phenolic reagents to form colored
compounds. Results and Discussion: Optimization of the formation of colored derivative compounds was carried
out by hydrolysis of guar gum with the addition of 1.0 mL of sulfuric acid, 30 min time, 100°C temperature, and color
stability at 180 min. Method validation was successfully carried out by testing several parameters, including selectivity
with the selected wavelength 492 nm. Linear regression equation y = 0.0255 × + 0.0420 (r: 0.9998) and Vxo = 0.76%.
The precision expressed in the Coefficient of Variation (C.V) is 1.06%. Accuracy was obtained 100.35 ± 0.56 (%; w/w).
Conclusion: In determining the galactomannan content in guar gum powder, it was 70.00 ± 0.24 (%; w/w).