Measurement of 3-Hydroxyisovaleric Acid in Urine of Biotin-Deficient Infants and Mice by HPLC1

• Published in: The Journal of nutrition Vol. 135; no. 3; pp. 615 - 618
• Main Authors: Watanabe, Toshiaki, Oguchi, Ken-Ichi, Ebara, Shuhei, Fukui, Toru
• Format: Journal Article
• Language: English
• Published: Bethesda Elsevier Inc 01.03.2005; American Institute of Nutrition
• Subjects: 3-hydroxyisovaleric acid; Acids; biotin deficiency; HPLC; methylcrotonyl CoA carboxylase; Urinalysis; Urine; Vitamins; 3-HIA; RT; methylcrotonyl CoA carboxylase; urine; 3-hydroxyisovaleric acid; biotin deficiency; MCC; 2-NPH; HPLC
• ISSN: 0022-3166; 1541-6100
• DOI: 10.1093/jn/135.3.615

We developed an assay for measuring urinary 3-hydroxyisovaleric acid (3-HIA) using HPLC after derivatization with 2-nitrophenylhydrazine hydrochloride (2-NPH · HCl). The derivatized 3-HIA was extracted into n-hexane and separated isocratically on a C8 reversed-phase column for fatty acids (YMC-Pack FA). We used this method to measure 3-HIA in urine extracts from mice fed a biotin-deficient diet for >4 wk and in an infant who was fed a special Japanese formula and was suspected of being biotin deficient. Urinary 3-HIA could be assayed within the range of 0.42–8.5 mmol/L with high accuracy by this method, as an indicator of biotin deficiency. Therefore, the HPLC method for 3-HIA described here may be a useful tool clinically as well as in the research laboratory.