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OBJECTIVE: To determine β2-microglobulin levels in amniotic fluid during the course of second trimester. STUDY DESIGN: One hundred patient’s amniotic fluid β2-microglobulin levels had been evaluated retrospectively (March-October 2009). The most common amniocentesis indication was advanced maternal age (33.3%). Others were; high risk result for triple test (18.5%), high risk result for double test (6.48%), ventriculomegaly (5.55%), obstetric history for fetus with down syndrome (4.62%), the presence of soft markers on ultrasound (13.8%), others (17.8%). Patients average gravida was 2.66 (range: 1-6), parity was 0.75 (range: 0-3), abortion was 0.65 (range: 0-3). RESULTS: All patients were at second trimester and the average gestational week was 17.7 (range 15- 22). Patients were divided into four groups (15th, 16th, 17-18th and 19-20th gestational weeks). We have demonstrated that amniotic fluid β2-microglobulin levels are increased progressively throughout the second trimester. We have specified normal β2-microglobulin values of each gestational week/period in order to be used in clinical practice. CONCLUSION: We have demonstrated that amnionic fluid β2-microglobulin levels are increased progressively throughout the second trimester.