Present study involves determination of arsenic in drinking water samples from different tehsils of district Sheikhupura along with the analysis of hair and blood samples of females of 15-25 years of age. Biological samples of males of same age group were also analyzed as control. Blood hormonal level of LH and FSH was measured in both groups whereas estradiol and testosterone was detected in females and males respectively using ELISA technique. AAS method was employed to measure arsenic in water, hair and blood. Questionnaire was designed for assessment of socioeconomic and general health status of the study group. The data was statistically analyzed for comparison and correlation. Majority of respondents belonged to lower socioeconomic group and had different health problems. Among 500 drinking water samples only 8.6% were within the WHO limit and only 50.6 % of the total samples were within NDWQS limit. The drinking water of tehsils Sheikhupura and Sharaqpur had higher arsenic as compared to other tehsils (64.25±2.55 µg/L and 61.63 ± 2.73µg/L) respectively and was highest in all hand pump water (71.14± 2.6µg/L). Arsenic concentration in hair was highest in 23-25 years females (1.62±0.10µg/g) and among the residents of tehsil Sheikhupura (1.40±0.12 µg/g) and those using hand pump water (1.31±0.14 µg/g). Similarly the highest mean value of blood arsenic was observed in the age group of 23-25 years (3.2 ±0.23 µg/L) and being highest among respondents of tehsil Sheikhupura. The same trend was noted in male group. Positive correlation (r=1.0) was found between arsenic in drinking water and biological samples with respect to area, age and water sources. In general FSH level in females was within reference range whereas LH was found in lower concentration in some areas. Estradiol was lower in follicular and luteal phase of the respondents from Sharaqpur and Sheikhupura suggesting possible exposure of arsenic. LH and FSH in males were in normal range and testosterone showed variability with reference to age. The results of this study evidently suggest that the presence of arsenic in drinking water is likely to affect general metabolism and its accumulation in biological tissues. This appears to be linked with the exposure of variable magnitude and duration. The results of this study provide enough data for future studies regarding water contamination and health effects also warrant urgency for preventive measures to reduce possible effects.