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Introduction Maternal nutritional status is considered by many Authors to be closely related to the reproductive success. The human reproductive process, from a probabilistic point of view, is ineffective and, in spite of the scientific research progressively adding elements to the understanding of reproductive failure, the mechanisms underlying the failure to conceive, the embryonic arrest and the occurrence of neonatal malformations remain largely unknown. In this area, nutrition and micronutrient status represent an important research target for two main aspects: 1) nutrition as a cause of reproductive failures 2) nutritional status as a key variable influencing the chance of success in assisted reproduction procedures. Aim This PhD project has been focused on the study of specific analytes in infertile couples, with particular reference to the probability of success in terms of pregnancy given particular conditions of deficiency / insufficiency. Methods and results The project was conducted at the ?Infertility Unit? of the Fondazione IRCCS Ca 'Granda Ospedale Maggiore Policlinico of Milan in collaboration with the Department of Clinical Sciences and Community at the University of Milan, in order to assess the baseline status of specific micronutrients and place it in relation to the chances of getting pregnant using assisted reproductive technologies. Evaluated analytes (vitamin D, folate, homocysteine, vitamin B12, vitamin E, vitamin E, iron and ferritin) were selected based on their involvement in the reproductive process. Subjects were recruited from couples treated for infertility; in the study of serum levels in infertile compared to subfertile women, recruitment was performed among pregnant women during the first trimester prenatal screening (U.O. Obstetrics and Gynecology). Several outcomes were evaluated using case series disjointed or partially overlapping. In particular, the PhD work was divided into 5 main studies, the characteristics of which are summarized in the following scheme: Study: 1 Analyte: Vitamin D Main Outcome: Pregnancy Rate in in vitro fertilization cycles Design: Prospective cross-sectional Subjects: 480 women in infertile couples Main results: Women with 25 (OH) D ? 20 ng / ml are more likely to have a pregnancy compared to those with 25 (OH) D insufficiency. Adjusted OR = 2.15 (95% CI: 1.23-3.77) (p = 0 .007). Study: 2 Analyte: Vitamin D Main Outcome: Concordance of serum levels between partners Design: Prospective cross-sectional Subjects: 103 infertile couples Main results: 71% of infertile couples showed concordance between a condition of sufficiency or insufficiency between partners. This frequency is higher than expected in case of random assortment between partners(p = 0.007). Study: 3 Analyte: Vitamin D Main outcome: Serum levels in fertile versus subfertile women Design: Prospective case-control Subjects: 73 cases and 73 controls Main results: Vitamin D status does not affect the natural fertility. OR for subfertility (time to pregnancy> 12 months) in women with low vitamin D = 0.85 (95% CI: 0.44-1.62) Study: 4 Analytes: Folate, Homocystein, Vitamin B12, Vitamin A, Vitamin E, Iron, Ferritin Main outcome: Rate of insufficiency Design: Prospective cross-sectional Subjects: 269 women in infertile couples Main results: Only a minority of women attending an Infertility Unit show pre-gestational values of ??intraerythrocytic folate (12%) and vitamin B12 (44%) Study: 5 Analytes: Folate, Homocystein, Vitamin B12 Main outcome: Pregnancy Rate in in-vitro fertilization cycles Design: Prospective cross-sectional Subjects: 209 women in infertile couples Main results: Women with serum and intraerythrocytic folate in the third tertile of concentration are more likely to obtain a clinical pregnancy in in-vitro fertilization cycles. OR = 2.8 (1.5-5.3) and 2.1 (1.1-4.0), respectively. Conclusion Infertile couples, and women in particular, have a high incidence of vitamin D, folate and vitamin B12 insufficiency. This condition, with specific regards to vitamin D and folate, while represents a risk factor for complications of a possible pregnancy, is strongly associated with a lower success rate of in-vitro fertilization. Further studies are needed to identify possible causal relationships between insufficiency of specific analytes and reproductive failure. The demonstration of a benefit in terms of pregnancies per in-vitro fertilization cycle through the use of supplementation of micronutrients would be a result of huge interest because it would represent the rationale for an effective, economic and easy intervention, to be implemented in an area where progress is slow and mostly dependent on expensive technologies.