Objective Neonatal hypoglycemia is the most frequent metabolic disorder in newborns and may result in permanent brain damage. There are insufficient valid studies to develop an evidence based guideline.<cross-ref type="bib" refid="b1">1</cross-ref> This may result in a wide variation in clinical guidelines. We analyzed currently used guidelines for neonatal hypoglycemia in The Netherlands. Methods National survey on guidelines for neonatal hypoglycemia in Dutch university and teaching hospitals. Results Response rate was 6/8 university hospitals (75%) and 20/27 teaching hospitals (74%). The cut-off for hypoglycemia was a glucose concentration <2.6 mmol/l in 81% of the hospitals; <2.0–2.2 mmol/l in 15%; one hospital used two cut-off values in the first 24 hours. Glucose concentrations were checked routinely in ‘high-risk’ newborn infants: median number: 3 times (2x–8x) for a median duration of 12 hours (3–72 hrs). In addition, different definitions were used for ‘high-risk’ infants, especially for small- or large-for-gestational-age infants: SGA: P10 in 50%, P2.3 in 15%, other: in 35% of the hospitals. LGA: P90 in 38%, P97.7 in 31%, other: in 31% of the hospitals. Conclusions There is considerable variation between Dutch hospitals in indication, timing and duration of routine glucose measurements in newborns at risk for hypoglycemia, reflecting the lack of evidence for currently used clinical guidelines. A Dutch multi-center trial has started comparing an intensive versus an expectant treatment strategy for moderate neonatal hypoglycemia ( www.neonatologiestudies.nl/hypoexit ).