The present study evaluated metabolic changes in lambs during three brief periods of ewe–lamb separation, namely 1 min (T1), 10 min (T10) and 60 min (T60). The lambs were divided by age into the following three groups: 15 days (G1 = 40 lambs), 30 days (G2 = 40 lambs) and 60 days (G3 = 40 lambs). The hypothesis to be tested was that breaking the ewe–lamb bond at different ages and for these brief time periods will cause significant metabolic changes in the lambs. To this end, blood samples were taken from the three test groups 24 h before separation to establish reference values (RVs), and again at the end of the three separation times. The following blood variables were analysed for all three study groups: glucose (mg/dL), lactate (mg/dL), calcium (Ca2+, mmol/L), sodium (Na+, mmol/L), potassium (K+, mmol/L), haematocrit (HTC, %), pCO2 (mmHg), pO2 (mmHg), HCO3– (mmol/L) and pH. Results showed that the G1 lambs suffered the greatest metabolic changes for the variables pO2, pCO2, glucose, lactate, pH, HCO3–, Ca2+, Na+, K+ and HCT. With respect to time, the first 10 min of separation from the mother induced marked metabolic changes in the 15-, 30- and 60-day-old lambs, but, at 60 min, the G2 and G3 lambs tended to recover their physiological RVs, while the G1 lambs did not. These findings showed that breaking the ewe–lamb bond immediately generated metabolic changes in the lambs even though they remained in a familiar, habitual environment. They also indicated that the age of the lambs at separation was a significant factor, as the older animals tended to re-establish their RVs after the metabolic changes, likely because their physiological compensating mechanisms are more efficient, or perhaps because the intensity of attachment to the mother decreases with greater age.