Pregnancy though a normal physiological process exposes the person to oxidative stress. Glutathione is a master antioxidant; it protects both mother and foetus from the damage of oxidative stress and free radicals. In the beginning of pregnancy, Glutathione protects essential functions of the growing and developing embryo by controlling cell differentiation, cell death and other essential functions in the developing embryo. Hence this study proposed the level of glutathione in the pregnancy. Aim: To study the glutathione level in pregnant women (2ndTRIMESTER) and to compare with the non-pregnant woman. Materials & Methods: 50 pregnant women (2nd TRIMESTER) and 50 non-pregnant women attending OBG department in government medical college, chennai Specimen: whole blood with EDTA. Method: Glutathione is determined by dithiobis nitro benzoicacid Glutathione levels were measured using the dithiobisnitrobenzoic acid (DTNB). Results: It was found that the glutathione levels in second trimester of pregnancy were found to be less than non-pregnant women. Conclusion: The result shows that oxidative stress during pregnancy is responsible for the reduction in glutathione as there is demand for the antioxidant glutathione. This study can be used to reinforce the necessity for the supplementation with dietary antioxidants like glutathione, vitamin C, vitamin E etc. Keywords: Pregnancy, Glutathione, Dithobisnitrobenzoic acid (DTNB).