Combur10test m stick

Urinalysis was performed for 5 rats in each group on week 13 of administration. Fresh urine (sampled within approximately 3 hr of urine excretion) was collected to determine pH, protein, glucose, ketone body, bilirubin, and occult blood using a urine analyzer (Miditron^® Junior II, Roche Diagnostics, Mannheim, Germany) or test kits (Combur 10 Test^®M stick; Roche Diagnostics). Urine was also visually observed for color and turbidity and the sediment was examined microscopically (Olympus BX50, Olympus Optical Co., Tokyo, Japan). In addition, 24-hr urine (after excretion) samples were examined for urine volume, specific gravity, sodium, and potassium using a measuring cylinder, a specific gravimeter (Vet360, Reichert, Depew, NY, USA), or an automatic analyzer (AVL9181, Roche Diagnostics, Mannheim, Germany).

On day 90, fresh urine was collected for 3 h from 5 rats of each sex per group in the fasting state, and a 21-h urine sample was subsequently collected while allowing access to food. Rats in metabolic cages (Biotoxtech, Cheongju, Republic of Korea) were allowed free access to water during both urine collection periods. The urine samples were stored at4°C until analysis was performed within 2 h of sample collection. The Combur¹⁰Test^®M stick (Roche Diagnostics, Mannheim, Germany) was used to measure pH, protein, glucose, and occult blood, while urine chemistry was done with a Cobas u 411 urine analyzer (F. Hoffmann-La Roche, Basel, Swizerland). Urine color and turbidity were checked by visual inspection. The 24-h urine was used for measurement of the urine volume and specific gravity. The specific gravity was measured with a Gravimeter (VET 360, Reichert Technologies, New York, USA).