Objective To assess ampicillin levels according to the duration of intrapartum antibiotic prophylaxis (IAP).

Design Prospective cohort single-centre study.

Setting Tertiary care centre (Modena, Italy).

Patients 120 neonates≥35 weeks’ gestation exposed to IAP.

Interventions Neonates were divided into four groups, according to the duration of IAP prior to delivery: group 1 (n=30; <1 hour), group 2 (n=30; ≥1 and <2 hours), group 3 (n=30; ≥2 and <4 hours) and group 4 (n=30; ≥2 doses, ≥4 hours).

Main outcome measures Blood samples were collected at delivery (from the umbilical cord) and at age 4 hours (from a peripheral vessel).

Results Median duration of IAP was 121 min (range 7–2045 min). Median ampicillin levels in umbilical cord blood were 10.4 µg/mL (IQR 6.4–14.9) and in peripheral blood were 4.7 µg/mL (IQR 2.8–6.4µg/mL). Umbilical cord blood levels reached a peak approximately 30 min after IAP and then declined significantly (p<0.001). Peripheral blood levels did not differ among study groups. Neonates exposed to a full loading dose (n=115) had peripheral blood levels 2.5–70 times higher than the minimal inhibitory concentration for group B streptococcus. There was no relationship between neonatal ampicillin concentrations and the duration of IAP prior to delivery (β=−0.0003, 95% CI −0.02 to 0.001, p=0.680).

Conclusions Ampicillin levels reach a peak in the umbilical cord blood within 30 min of intrapartum administration. After a full loading dose, bactericidal levels persist for at least 4 hours after birth and seem independent of the duration of IAP prior to delivery.