Background: Reference values for serum Mg in adults are well defined but in paediatric patients and more specifically in newborn and preterm infants (lesser than 37 weeks gestation) reference values are either not available or are very limited and still controversial; most laboratories do not provide specific reference intervals for this population. Most of studies are done in preterm newborns exposed to prenatal MgSO4 and few studies done in preterm newborns who are not exposed to prenatal MgSO4 However there is no conclusive study from developing countries which elicits if the same reference interval is equally effective in the Indian population. The Indian women differ from those in developed country in terms of constitution, body surface area etc. which may influence the pharmacodynamic action and its effects in neonates. Aim: To assess serum magnesium levels in preterm newborn who were not exposed to prenatal MgSO4. Objective: We have done study to find out the normal serum magnesium values in preterm neonates at birth and to establish the relationship, if any, of these biochemical values with birth weight, gestational age and maternal BMI. Methods: Present prospective observational study was conducted at Level III b NICU of the Department of Paediatrics and Neonatology, Deenanath Mangeshkar Hospital and Research Centre Pune between January 2022 to December 2022. One hundred sixteen preterm newborns (lesser than 37 weeks of gestational age) were studied to estimate the normative data of serum magnesium level in preterm neonates not exposed to prenatal MgSO4 and its variation with gestational age and weight and maternal BMI. Results were analysed and interpretated. Results: In the present study the mean values of serum magnesium irrespective of birth weight, gestational age and sex were observed as –Serum magnesium 1.97±0.38 mg/dl. Conclusion: In conclusion, our findings add to the evidence that the there is consistent variation of serum magnesium level of preterm neonates who were not exposed to antenatal MgSO4 with respect to birth weight and gestational age and showed inverse relation with both factors. There was no correlation found between gender of babies and maternal BMI.