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Twenty-four-hour urine specimens from 21 juvenile insulin-dependent diabetics and 10 healthy controls were compared with respect to biotinidase activity and alanine content. Urinary biotinidase activity was analysed by a newly developed high-performance liquid chromatography (HPLC) method. It was found that the excretion of biotinidase in urine was elevated in diabetics (7.02 mU/d; p < 0.005) as compared with controls (not detectable). Alanine excretion was also found to increase (p < 0.01) in diabetics. Biotinidase excretion in diabetics was correlated with alanine excretion (rS = 0.667; p < 0.01), but not with protein, albumin or N-acetyl-glucosaminidase excretion. The simultaneous elevation of urinary biotinidase and alanine excretion in juvenile diabetics suggests that changes in kidney metabolism arise in the early stages of diabetes.

Journal of Liquid ChromatographyPublication details, including instructions for authors and subscription information:ABSTRACT Sensitive high-performance gel-permeation chromatographic protein assay method was developed by using non-ionic detergent of Brij-58 which was UV transparent as compared to Nonidet P -4 0 . A column (70 x 8.0 mm I.D.) packed with Develosil 100 Diol-5 ( p o r e size 10 nm) was used for the protein determination. A commercially available column (300 x 8.0 mm I.D.) packed with Develosil 300 Diol-5 (pore size 30 nm) was used for the urinary albumin determination.Eluent used was a 0.1 M sodium phosphate buffer (pH 5 . 6 ) containing 0.3 M sodium chloride, 1% (v/v) Brij-#A part of this vork was presented in the 20th annual meeting of the international society for pediatric and adolescent diabetes, Atami-city, Shizuoka, Japan, November 1994. 3955Downloaded by [McGill University Library] at 09:18 07 February 2015Flow-rates for protein assay and albumin assay were 1 . 0 and 0.5 ml/min, repectively.Proteins were eluted at the position of the exclusion limit in the case of protein assay ( 1 0 0 Diol-5).Albumin was eluted from 300 Diol-5 column as a symmetric peak.Bovine serum albumin (BSA) was used as an external standard f o r both protein and albumin assays.Analysis times were within f o u r min for protein assay, and 32 min for albumin assay, respectively. Improved sensitive measurement of BSA at 5-20 ng level was achieved as compared to Nonidet P-40 by use of UV 210 nm.This method was successfully applied f o r various urine samples, such as healthy random urine of before and after sports, and the 24-h urines from insulindependent diabetes mellitus (IDDM) patients.Comparisons with clinical urinary protein and albumin tests were performed using IDDM urine.T h r e e cases out of four tests for sports showed increased protein and albumin content after physical exercise. Thus, this HPLC method was proven to be applicable to the protein and albumin measurements in human urine.