d-Lactate has been proposed as biomarker for subacute ruminal acidosis (SARA). This study aimed to evaluate d-lactate absorption and metabolism in dairy cows under high or low risk for SARA. Six lactating Holstein dairy cows fitted with ruminal cannula and indwelling catheters in major splanchnic blood vessels were used. Animals were randomly assigned to either a SARA diet (80:20 concentrate to forage; 370 g starch/kg dry matter) or a control diet (50:50 concentrate to forage; 225 g starch/kg dry matter) in a cross over design with 28-d periods. On d 20, 8 hourly sample sets of fluid from the medial and ventral rumen, and blood from an arterial, portal and hepatic vein were collected, starting 30 min after the morning feeding. After 4 h relative to feeding, 1.75 kg of ground barley was administrated via the ruminal cannula in all cows. Net D-Lactate releases from the portal drained viscera (PDV) and the total splanchnic tissues (TSP) were calculated. The MIXED procedure of SAS was used considering sampling time as repeated measure. No differences were detected between groups for DMI (20.3 ± 0.8 kg/d; P > 0.05) and milk yield (30.5 ± 1.2 kg/d; P > 0.05). In the first 4 h after feeding, ventral rumen pH did not differ between treatments (P > 0.05). Medial rumen pH and d-lactate concentration in ventral and medial rumen constantly decreased in both groups after morning feeding (P < 0.05), but did not differ between treatments (P > 0.05). Arterial d-lactate as well as the net PDV release of d-lactate decreased after feeding (P < 0.05), but did not differ between treatments (P > 0.05). The d-lactate release from TSP decreased in both groups after feeding (P < 0.05), but did not differ between treatments (P > 0.05). The direct rumen administration of ground barley at 4 h after feeding increased d-lactate release from TSP in both groups (P < 0.05), but the increase did not differ between treatments (P > 0.05). The other variables were not affected by either time after administration or diet (P > 0.05). In conclusion, ruminal d-lactate absorption and metabolism did not differ in cows fed the SARA diet compared with the control. The lack of differences between experimental groups may indicate adaptation of the microflora to high starch diets.