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Multivitamin and mineral (MVM) supplements are frequently used amongst older populations to improve adequacy of micronutrients, including B-vitamins, but evidence for improved health outcomes are limited and deficiencies remain prevalent. Although this may indicate poor efficacy of supplements, this could also suggest the possibility for altered B-vitamin bioavailability and metabolism in older people. This open-label, single-arm acute parallel study, conducted at the Liggins Institute Clinical Research Unit in Auckland, compared circulatory and urinary B-vitamer responses to MVM supplementation in older (70.1 ± 2.7 y, <i>n</i> = 10 male, <i>n</i> = 10 female) compared to younger (24.2 ± 2.8 y, <i>n</i> = 10 male, <i>n</i> = 10 female) participants for 4 h after the ingestion of a single dose of a commercial MVM supplement and standardized breakfast. Older adults had a lower area under the curve (AUC) of postprandial plasma pyridoxine (<i>p</i> = 0.02) and pyridoxal-5′phosphate (<i>p</i> = 0.03) forms of vitamin B₆ but greater 4-pyridoxic acid AUC (<i>p</i> = 0.009). Urinary pyridoxine and pyridoxal excretion were higher in younger females than in older females (time × age × sex interaction, <i>p</i> < 0.05). Older adults had a greater AUC increase in plasma thiamine (<i>p</i> = 0.01), riboflavin (<i>p</i> = 0.009), and pantothenic acid (<i>p</i> = 0.027). In older adults, there was decreased plasma responsiveness of the ingested (pyridoxine) and active (pyridoxal-5′phosphate) forms of vitamin B₆, which indicated a previously undescribed alteration in either absorption or subsequent metabolic interconversion. While these findings cannot determine whether acute B₆ responsiveness is adequate, this difference may have potential implications for B₆ function in older adults. Although this may imply higher B vitamin substrate requirements for older people, further work is required to understand the implications of postprandial differences in availability.