Urinary 3HIA and 3HIA-carnitine responses to leucine challenge (A,C) and in the preceding 24-h urine samples (B,D) increase with biotin depletion in 5 healthy adults (Study 1). The gray rectangle denotes the NR. Solid bars denote mean excretion at d 0 and 28. Asterisks indicate that the mean on that day differs from d 0: *P < 0.02, **P < 0.002, ***P < 0.0005. 3HIA, e-hydroxyisovaleric acid; 3HIA-carnitine, 3-hydroxyisovaleryl carnitine; NR, normal range.