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MB Sample ID: SA005253
Local Sample ID: | ConstantVolume_WT_2_2 |
Subject ID: | SU000111 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Male and Female |
Human Race: | Hispanic, Aisan, Caucasian |
Species Group: | Human |
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Treatment:
Treatment ID: | TR000112 |
Treatment Summary: | Varying Volume with Urease / Male vs. Female with Urease / Constant Volume with Urease / Constant Volume with 100 µL Water |
Treatment Protocol Comments: | To evaluate whether the effects of urease pre-treatment of urine varied with the volume of urine prepared, we compared the urine metabolite profiles from pooled urine after pre-treatment with urease (UT) and after no treatment (NT) using various volumes of urine. For this, several volumes (5, 10, 25, 50, and 100 µL) of the pooled urine sample were incubated (n = 3, each) with 100 µL of a 1 mg/mL solution of urease or were not subjected to any treatment. / Finally, to evaluate whether any artifacts introduced by urease pre-treatment on the urinary metabolome interfered with the ability to distinguish between comparative samples, we compared the metabolite profiles from individual male and female urine samples after pre-treatment with urease or after no treatment (previous metabolomics studies of male and female urines (Pasikanti et al., 2008; Slupsky et al., 2007; Saude et al., 2007; Psihogios et al., 2008) have reported differences in metabolite levels). For this, 50 µL aliquots of individual male and female urine samples (n = 20, each) were blocked, randomized, and then incubated with 50 µL of a 1 mg/mL solution of urease (UT) or were not subjected to any treatment (NT), each as described above. / To initially evaluate the effects of urease pre-treatment on the urinary metabolome, we compared the urine metabolite profiles from pooled urine after pre-treatment with urease, water, or no treatment at all. For this, 100 uL aliquots of the pooled urine sample were incubated (n = 5, each) with 100 uL of a 1 mg/mL solution of urease (Sigma-Aldrich catalog number U4002) prepared in water (urease-treated; UT) or an equal volume of water alone (water-treated; WT) for 30 min at 37 C with mild shaking (500 rpm). Identical aliquots (n = 5) were not subjected to any treatment (no treatment; NT) and allowed to sit at room temperature for 30 min. / To initially evaluate the effects of urease pre-treatment on the urinary metabolome, we compared the urine metabolite profiles from pooled urine after pre-treatment with urease, water, or no treatment at all. For this, 100 uL aliquots of the pooled urine sample were incubated (n = 5, each) with 100 uL of a 1 mg/mL solution of urease (Sigma-Aldrich catalog number U4002) prepared in water (urease-treated; UT) or an equal volume of water alone (water-treated; WT) for 30 min at 37 C with mild shaking (500 rpm). Identical aliquots (n = 5) were not subjected to any treatment (no treatment; NT) and allowed to sit at room temperature for 30 min. |
Treatment: | Abiotic |
Treatment Compound: | Urease / Urease / Urease / Water |
Treatment Dose: | 1 mg/ml / 1 mg/ml / 1 mg/ml / -- |
Treatment Dosevolume: | 100 µL / 50 µL / 100 µL / 100 µL |
Human Fasting: | Overnight Fast |