AimTo investigate the clinical characteristics and course of parenteral nutrition‐associated cholestasis (PNAC) in very low birth weight (VLBW) infants.MethodsThe charts of VLBW infants were retrospectively reviewed. The clinical characteristics of infants with and without PNAC were compared, trends in liver enzymes were investigated, and the characteristics of infants with PNAC were analysed based on age of onset.ResultsPNAC was observed in 53 (13.2%) of 403 infants who survived and completed follow‐up and was associated with significantly lower gestational age, birth weight, and adverse neonatal outcomes. PNAC started at a median 32 (interquartile range 23–47) days, PN was applied for 53 (34.5–64.5) days, the maximum direct bilirubin (DB) was observed at 63 (50–76) postnatal days, and PNAC resolved at 94 (79–122) postnatal days postnatal age. PNAC lasted 61 (38–89.5) days. AST and ALT normalised at 111 (100.3–142.0) and 109.5 (97–161.3) postnatal days. Infants with early‐onset PNAC had significantly longer PN duration, higher maximum DB, and higher maximum AST than those with late‐onset PNAC.ConclusionElevated DB, AST, and ALT persist for a long period after discontinuing PN. We suggest a cautious approach that involves waiting and reducing the frequency of additional repetitive examinations.