Spectrum subtraction as a complementary method for six resolution techniques resolving overlapping spectra;...

Area of Science:

• Analytical Chemistry
• Spectrophotometry
• Green Analytical Chemistry

Background:

• Overlapping spectra in binary mixtures pose analytical challenges.
• Spectrophotometric methods offer economic and precise analysis of pharmaceutical compounds.

Purpose of the Study:

• To compare six economic, accurate, and precise spectrophotometric methods for Triclabendazole (TCB) and Levamisole HCl (LVM) determination.
• To evaluate mathematical filtration techniques in zero and first-order spectra for resolving binary mixtures.
• To assess the greenness of the developed analytical methods.

Main Methods:

• Comparative analysis of six spectrophotometric methods: ratio subtraction, constant multiplication, factorized response, and spectrum subtraction in zero and first derivative orders.
• Determination of linearity range, Limit of Detection (LOD) for TCB and LVM.
• Validation according to VICH guidelines.
• Green assessment using NEMI, AES, GAPI, and AGREE tools, plus RGB model white assessment.

Main Results:

• All proposed methods demonstrated high accuracy and precision, comparable to reported techniques.
• Linearity ranges were established for TCB (1.0-10.0 µg/mL or 2.0-20.0 µg/mL) and LVM (2.0-14.0 µg/mL).
• LOD values were 0.08 µg/mL for TCB and 0.19 µg/mL for LVM.
• The developed methods align with green analytical chemistry principles.

Conclusions:

• The study successfully resolved TCB and LVM in binary mixtures using various mathematical filtration techniques.
• The proposed spectrophotometric methods are economic, accurate, and precise alternatives for routine analysis of bulk materials and veterinary formulations.
• The green assessment confirms the environmental viability of the developed analytical procedures.