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MB Sample ID: SA002203
| Local Sample ID: | UPALL0001 |
| Subject ID: | SU000068 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | 24 female; 16 male |
| Species Group: | Human |
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Sample Preparation:
| Sampleprep ID: | SP000064 |
| Sampleprep Summary: | Frozen urine study samples were thawed on ice and vortexed for 30 seconds. Each of the 40 urine samples (200 µL) were prepared individually by addition of 25 µL D20 and 25 µL Chenomx Internal Standard (Chenomx ISTD, Edmonton, Alberta, Canada). Phenotypic pools for AKI and no AKI were generated with 60 µL of each urine sample pooled, mixed, divided into 3 aliquots. Additionally, 400 µL from both phenotypic pools were mixed to form a total pooled sample and divided into 3 aliquots. The pooled samples were prepared identically to the individual study samples. 1H NMR spectra of urine samples were acquired on a Bruker Avance 950 MHz NMR spectrometer (located at the David H. Murdock Research Institute at Kannapolis, NC, USA) using a 3 mm cryogenically cooled ATMA inverse probe and ambient temperature of 25℃. A 1D NOESY presaturation pulse sequence (noesypr1d, [recycle delay (RD)-90°-t1-90°-tm-90°-acquire free induction decay (FID) was used for data acquisition. For each sample 64 transients were collected into 65k data points using a spectral width of 14.01 kHz (20.14 ppm), 2 s relaxation delay, 100 ms mixing time, and an acquisition time of 2.324 s per FID. The water resonance was suppressed using resonance irradiation during the relaxation delay and mixing time. NMR spectra were processed using Chenomx NMR Suite 7.51 Professional (Chenomx, Edmonton, Alberta, Canada) software. Spectra were zero filled, and Fourier transformed after exponential multiplication with line broadening factor of 0.5. Phase and baseline of the spectra were manually corrected for each spectrum. Spectra were referenced internally to the DSS signal. The quality of each NMR spectrum was assessed for the level of noise and alignment of identified markers. Spectra were assessed for missing data and underwent quality checks. NMR bins (0.50-9.0 ppm) were made after excluding DSS, water (4.68-4.80 ppm), and Imidazole (7.20-7.28 ppm) using bucket Integration with a 0.04 ppm bucket width. Integrals of each of the bins were normalized to total integral of each of the spectrum. |
| Sampleprep Protocol Filename: | RTI_NeonatalAKI_Metabolomics_Procedure.docx |
