Currently, Whatman 903 (W-903) filter paper is the only filter paper that has been used for collection of blood samples in thyroid hormones. This commercially available filter paper has so expensive which is not suitable for epidemiological study conducted in developing countries. We therefore introduced a new matrix for collection of blood samples and also compared with commercial available matrix, Whatman filter paper (W-903). We used two matrices initially for standardisation purpose viz., the conventional Whatman dbs cards (#903 10531018) & pre coated TLC silica gel matrix (60F254, Merck 20x20 Cm), the latter with cost-effectiveness in view. Both were subjected to validation for optimum working conditions. The assay characteristics for Whatmann DBS card and TLC matrix were found to be as follows: correlation coefficient 0.982 & 0.976, slope 0.070 & 0.068, coefficient of variation 0.32-0.77% & 0.40-0.85%, sensitivity 0.0287 & 0.0286 mIU/L and interassay variation 0.40% & 0.50 respectively. The assay characteristics of the calibration curve for FT4: correlation coefficient 0.926 , slope 0.094, coefficient of variation 0.84-4.12% and inter assay variation 3.03% for TLC silica matrix. Drying time (0.5, 1.5, 2.5, 3.5 , 4.5 ,5.5 h), elution time (1.0, 3.0, 5.0-16 h), percent recovery (83-98%) and linearity at varying dilutions were determined. Stability studies for TSH samples were conducted at variable temperatures and half life found to be temperature dependent viz.,24.54 days, RT; 180.45, 4 0 C; 206.25 days,-20 0 C. Owing to its cost-effectiveness and comparable results, TLC silica matrix was preferred for subsequent studies under optimized conditions.