Present research was carried out to evaluate the efficacy of psyllium husk as a therapeutic agent in normal and hypercholesterolemic subjects. For the purpose, dietetic cookies were prepared by replacing straight grade flour with psyllium husk in different combinations @ 5, 10, 15, 20 and 25%. Psyllium husk was analysed for chemical composition, dietary fiber through enzymatic method and functional component arabinoxylan (AX) using GC-MS. Characterization results indicated psyllium husk as a rich source of dietary fiber and arabinoxylan. The treatments of prepared cookies were analysed for physical, chemical and sensory attributes with special reference to dietary fiber, arabinoxylan, gross energy, hardness and color tonality. Physical characteristics showed decline in diameter and spread factor whilst, increase in thickness of treated cookies. Chemical assay revealed higher protein in control cookies whereas moisture, ash, crude fiber & NFE along with dietary fiber and arabinoxylan contents were higher in psyllium based cookies. Softer cookies with low gross energy and better color tone were formed using psyllium husk. Sensory panel explicated cookies quite acceptable however decrease in sensory score was recorded during storage. Considering the results of characterization, dietary fiber and sensory response, one best treatment namely T4 (20% replacement of psyllium husk) was selected along with control for two months efficacy study in human subjects during two consecutive years i.e. 2009-10 (Trial-I) & 2010-11 (Trial-II) for validity of the results. Bio-evaluation trial was carried out in normal subjects following same module in hypercholesterolemics by providing five cookies twice a day to each subject. Results indicated non-significant differences in serum lipid profile and glycemic indices (glucose and HbA1c) of normal subjects after consuming dietetic cookies (T4). Nevertheless, psyllium husk based cookies had positive impact to attenuate the lipid profile of hypercholesterolemic subjects. In this context, elevated base line values of total cholesterol 231.50±6.46, LDL 158.78±8.91, VLDL 35.92±1.36, LDL/HDL 4.99±0.25 and glucose 103.10±7.90mg/dL, were significantly reduced to 202.90±7.26, 134.60±8.13, 30.40±1.05, 4.09±0.17 and 92.07±4.11mg/dL, respectively in the Trial-I. Likewise, significant declining trend was observed in respective traits due to treatment T4 in Trial-II. The percent reduction in total cholesterol was 12.35 & 11.26%, LDL 15.22 & 14.10%, VLDL 13.48 & 12.08% and glucose 10.70 & 12.19%, respectively in Trial-I &II for hypercholestrolemic subjects due to dietetic cookies (T4). However, HDL concentration increased non-momentously during the study period. Likewise, triglycerides level was also affected non-significantly however, a slight decline was recorded. Safety evaluation of dietetic cookies was also carried out simultaneously in both categories of subjects and no sign of abnormality was observed in liver (ALT & AST) and kidney functions (urea & creatinine). It has been observed that high base line value for urea was non-significantly declined after administration of test diet in hypercholesterolemic subjects. Furthermore results for hematological tests depicted safe range in allied parameters i.e. platelets count, ESR, red blood cells and white blood cells indices. It is interesting to mention that the ESR of hypercholestrolemic subjects was momentously affected by using therapeutic diet (T4) as it was reduced to 5.61 and 6.98% in Trial-I & II, respectively. It has been observed that psyllium husk is friendly to the normal subjects nonetheless supportive for the hypercholesterolemic subjects. From the current explorations it is concluded that psyllium husk based cookies (supplemented with 20% psyllium husk) have potential to be used as a functional ingredient against the menace of hypercholestolemia and allied discrepancies in vulnerable segments of the population.