BACKGROUND:Pregnancy is a natural physiological statement with hormonal and metabolic changes that helps the growth and survival of the fetus. However, biochemical profiles derangement may lead to pregnancy complications. Therefore, there is a need for  determining biochemical profiles amongpregnant women. METHODS: A comparativecross-sectional study was conducted among pregnant and non-pregnant women at the University of Gondar Hospital,  from February to April, 2015. Fasting blood sample was collected from 139 pregnant and 139 age matched non-pregnant women using systematic random sampling technique. Interviewer-administered questionnaire was used to collect socio-demographic and clinical data. Fasting blood glucose and lipid profile were measured by A25 Biosytemchemistry analyzer using enzymatic calorimetric methods. Data analysis was done using SPSS version 20. Level of significance between groups was analyzed using independent student t-test and Mann-Whitney U test. A p-value of <0.05 was considered as statistically significant.RESULT: Pregnant women as compared to non-pregnant had significantly increased glucose (96.35+14.45 and 81.12+9.86 mg/dl), total cholesterol (211.9+40.88 and 172.40+29.64 mg/dl)[p<0.05], respectively. It had also significantlyhigh triglycerides (190.81+81.04 and 107.43+45.80 mg/dl) and low-density lipoprotein cholesterol (116.03+37.26 and 86.12+27.29mg/dl)[p<05] in pregnant as compared to non-pregnant women. The level of high-density lipoprotein cholesterol wassignificantly lower in pregnant women (59.58+14.26)than control (63.63+11.4, P <0.05).CONCLUSION: There were statistically significant increment in glucose, total cholesterol, triglycerides,  low-density lipoprotein cholesterol and decrement in high-density lipoprote in cholesterol levels among pregnant women compared with non-pregnant women. Therefore, pregnant women have to be monitored closely for their biochemical profiles to avoid adverse pregnancy outcomes. KEYWORDS: Pregnancy, biochemical profiles, Gondar, Ethiopia