Alveolar surfactant is well known for its ability to reduce minimal surface tension at the alveolar air-liquid interface to values below 5 mN m-1. In addition, it has been suggested that surfactant is also present in the airways, particularly in the perinatal period. We isolated surfactant from pharyngeal aspirates obtained from 33 neonates immediately after delivery and analysed it for both phospholipid (PL) composition and surface tension function. PL classes and phosphatidylcholine (PC) molecular species were determined by normal and reversed-phase high-performance liquid chromatography (HPLC), respectively. Static and dynamic surface properties of the surfactant were studied in a pulsating bubble surfactometer. Sample volume was 1.3 +/- 0.5 mL (mean +/- SD) with a total amount of 2.5 +/- 1.3 mumol of PL and a concentration of 2.1 +/- 1.0 mumol mL-1 PL. HPLC analyses of PL classes revealed a composition identical with surfactant prepared from alveolar washes, i.e. PC 83.6 +/- 2.1%, sphingomyelin 1.4 +/- 0.5%, phosphatidylglycerol 8.1 +/- 1.6%, phosphatidylethanolamine 2.1 +/- 0.5% and phosphatidylinositol 2.6 +/- 1.1%. Thin-layer chromatography showed almost identical results but was more time-consuming and needed more material for analysis. Analysis of PC molecular species revealed a composition typical of human alveolar surfactant with 54.7 +/- 3.9% dipalmitoyl PC, 10.3 +/- 1.9% palmitoyloleoyl PC and 9.1 +/- 1.5% palmitoylmyristoyl PC. Minimal surface tension fell to values below 5 mNm-1 within 5 min of cycling in all subjects. The methods used in this study allowed for complete PL and surface tension analyses of surfactant obtained during routine pharyngeal suctioning after delivery at term. Whether they are also applicable to preterm neonates with respiratory distress remains to be determined.