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We undertook this study to propose a noninvasive method for measuring oxytocin (OT) level in pregnant women. Most of previous studies have focused on the measurement of OT levels in blood plasma through invasive method. Due to the important role of OT hormone level during pregnancy, in this study, the level of salivary OT during pregnancy was measured to investigate its applicability as an alternative to invasive conventional methods. In this case-control study, 126 individuals (63 pregnant women as case group and 63 non-pregnant women as control group) referred to in Fatemiyeh Hospital of Hamadan, Iran, were selected and evaluated. After obtaining written consent, saliva sampling was performed by stripping, and the samples were transferred to a laboratory at -4°C and then stored at -22°C. ELISA technique was applied for measuring the level of salivary OT. Data were analyzed using SPSS software. The results indicated that the mean level of salivary OT in the control group was 98.40±362.92 pg mL-1. The mean level of salivary OT in the case group was 1.016±403.75 pg mL-1 with the maximum and minimum concentrations of 628.60 and 169.60 pg mL-1, respectively. The results of this study implied that the mean level of salivary OT in pregnant women was significantly higher than that in non-pregnant women (P = 0.024). Also, OT levels were not significantly different in trimesters of pregnancy (P1 = 0.941, P2 = 0.844, P3 = 0.552). Our findings depicted that measuring salivary OT in pregnant women can be used as a noninvasive and accurate method instead of blood test.