Wide Range of Biotin (Vitamin H) Content in Foodstuffs and Powdered Milks as Assessed by High-performance Affinity Chromatography

Abstract

The biotin (vitamin H) contents of various foodstuffs were determined by using a newly developed high-performance affinity chromatography with a trypsin-treated avidin-bound column. Biotin was derivatized with 9-anthryldiazomethane (ADAM) to fluorescent biotin-ADAM ester. A wide range of biotin contents were found in various foodstuffs depending upon the species (strain), season, organ (of plants and animals), geography, freshness, preparation method and storage method. Among the foodstuffs and fermented foods tested, it was found that wide distributions of biotin content were observed in powdered milk, natto, sake (rice wine), beer, edible oil and sea weed. Since powdered milk is important for child health and development, 14 kinds of powdered and special milks for use in children’s diseases were intensively measured. We found that several special milk powders for children with allergies contained low levels of free biotin. Use of these powdered milks caused skin diseases and alopecia in some patients possessing thermolabile serum biotinidase, and administration of free biotin improved their symptoms dramatically. Therefore, it is essential to estimate the total and free biotin contents on each foodstuff in order to improve effective biotin intake and support better health and quality of life for people.

Changes in the Hydrophobic Proteins in Response to Biotin Administration in Serum of Infant Patients with Alopecia as Assessed by the Protein Micro-Sequencing Method

“…With respect to the inflammation, the affinity to the substrate (biotinyl-6-aminoquinoline) of biotinidase (Amo) in inflammatory disease is also found to be significantly increased; i.e., median Amo in inflammatory diseases (n=10) is 23.9 s -1 x M -1 and that in healthy control (n=8) is 19.4 s -1 x M -1 , respectively (p<0.002; Mann-Whitney's U test, two-tailed test). This finding is in line with the previous finding that specific activity (V) of biotinidase is instantaneously elevated in cerebrospinal fluid of a Staphylococcus aureus meningitis patient [18]. Surprisingly, protein amount of biotinidase in the serum is not changed by the oral biotin-administration at all; i.e., Child patient 1 from 0.19 to 0.19 mg/mL, Child patient 2 from 0.20 to 0.22 mg/mL, and Child patient 3 from 0.25 to 0.21 mg/mL before and after the biotin treatment, respectively (Table 1 and Supplementary data 3).…”

“…Free biotin in the serum is increased for 7.3-fold during 1.5 y of this therapy. The symptoms of this patient seem to be slightly improved by this new therapy [18].…”

“…Serum total-and free-biotin were determined by the affinity-HPLC method as described previously [17,18].…”

“…Thermal instability test was also performed by incubating the diluted serum by the 0.1 M sodium phosphate buffer (1 mM EDTA, 10% (v/v) glycerol, pH 6.8 [13]) for 4 h at 37°C [18]. Effect of fucoidan and heparin were tested at 0.20 mg/mL in the reaction mixture.…”

“…Since the numbers of human sera were insufficient for estimating the distribution pattern and the biochemical phenomena possibly might have no parametric-distribution patterns, non-parametric analysis was performed as described previously [18]. Then, the data values were indicated as median and range.…”