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BACKGROUND: Pregnancy is a natural physiological statementwith hormonal and metabolic changes that helps the growth andsurvival of the fetus. However, biochemical profiles derangementmay lead to pregnancy complications. Therefore, there is a needfor determining biochemical profiles among pregnant women.METHODS: A comparative cross-sectional study was conductedamong pregnant and non-pregnant women at the University ofGondar Hospital, from February to April, 2015. Fasting bloodsample was collected from 139 pregnant and 139 age matchednon-pregnant women using systematic random samplingtechnique. Interviewer-administered questionnaire was used tocollect socio-demographic and clinical data. Fasting blood glucoseand lipid profile were measured by A25 Biosytemchemistryanalyzer using enzymatic calorimetric methods. Data analysis wasdone using SPSS version 20. Level of significance between groupswas analyzed using independent student t-test and Mann-WhitneyU test. A p-value of <0.05 was considered as statisticallysignificant.RESULT: Pregnant women as compared to non-pregnant hadsignificantly increased glucose (96.35+14.45 and 81.12+9.86mg/dl), total cholesterol (211.9+40.88 and 172.40+29.64 mg/dl)[ p<0.05 ] , respectively . It had also significantly high triglycerides(190.81+81.04 and 107.43+45.80 mg/dl) and low-densitylipoprotein 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 was significantlylower in pregnant women (59.58+14.26) than control (63.63+11.4,P <0.05).CONCLUSION: There were statistically significant increment inglucose, total cholesterol, triglycerides,low-density lipoproteincholesterol and decrement in high-density lipoprote in cholesterollevels among pregnant women compared with non-pregnant women.Therefore, pregnant women have to be monitored closely for theirbiochemical profiles to avoid adverse pregnancy outcomes.