Creatinine recovery from bovine urine under the effect of different times and temperatures of storage

Abstract

Creatinine is a urinary marker used widely in ruminant’s experimental trials. However, despite its great importance no data were found in the literature about the best way to store bovine urine samples. In the sheep urine, was observed an increase in the urinary concentration of creatinine when it was stored acidified (pH 2.5 to 3.5) at a temperature of 28 to 39 °C for 150 days of storage. Nevertheless, urine should be stored acidified (pH below 3) to avoid purine derivative degradation, So, aimed to evaluate creatinine recovery in bovine urine as a function of storage time and temperature. A total of 25 animals’ urine (10 Nellore cattle and 15 Holstein cattle) were collected. The urine (40 mL) was diluted in 160 mL of distilled water and its pH was corrected to a value lower than 3 using sulfuric acid drops. A sample of the diluted urine was analyzed to obtain the creatinine concentration reference value on the collection day. The remaining urine was fractionated and preserved at room temperature, cooled (4 °C) or frozen (-20 °C and -40 °C). In the urine of five Holstein cattle was added creatine solutions (20, 40 and 60 mg/dL) to evaluate the creatine to creatinine conservation. These urine samples were analyzed on different days after collection (1, 3, 7, 15, 30 and 45 days). The urine without any added creatine was analyzed on Days 1, 3, 7, 10, 15, 30, 45, 60, 90, 120, and 150 of storage. The addition of creatine in the urine caused an increase in the creatinine concentration (P < 0.05) after 30 days of storage at room temperature and under refrigeration (4 °C). In frozen samples, there was no change in creatinine concentration (P > 0.05). However, creatinine recovery was constant (P > 0.05) until day 15 of storage, regardless of the temperature used, when creatine was not added. After 30 days of storage, an effect of time and/or temperature was observed on creatinine recovery (P < 0.05). Urine samples can be stored at any temperature for up to 15 days after collection to estimate the creatinine concentration. Samples that need storage times longer than 15 days should be frozen (at -20 °C and -40 °C) to avoid creatinine concentration variation.