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MB Sample ID: SA323333
| Local Sample ID: | CSH_Blank_Neg_01 |
| Subject ID: | SU003090 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 61.1 +/- 15.2 |
| Gender: | Male and female |
| Human Lifestyle Factors: | Unsuccessful antibiotic treatment for recurrent C.difficile infection |
| Human Medications: | Vancomycin, Metronidazole, Fidaxomicin, probiotic |
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Sample Preparation:
| Sampleprep ID: | SP003096 |
| Sampleprep Summary: | Fecal samples were weighed into pre-tared 2 mL Precellys (Bertin Corp.) compatible vials, and one 2.8 mm stainless steel bead was added to aid homogenization. Fecal matter was homogenized using a Precellys Evolution by two 20 s cycles separated by a 30 s break. Extraction solvent was used at a ratio of 1 mL per 5 g feces and was comprised of 1:1:1 methanol:acetonitrile:acetone containing 10 uM of D3-creatine, D10-isoleucine, D2-biotin, D5-tryptophan, D3-caffeine, D3-octanoylcarnitine, D3-palmitoylcarnitine, D4-deoxycholic acid, D4-cholic acid, and D7-arginine as internal standards. Following extraction, samples were centrifuged for 10 min at 17,000 rpm. 100 uL aliquots of supernatant were transferred to Eppendorf vials, dried under a gentle stream of nitrogen, and stored at -80 ºC. On the day of analysis, the dried extracts were reconstituted in 85:15 acetonitrile:water for HILIC analysis or 9:1 water:methanol for RPLC analysis with volumes as described below. Pooled samples were prepared by combining equal volumes of reconstituted fecal matter extracts from all subjects. |
| Processing Storage Conditions: | On ice |
| Extraction Method: | 1 g feces / 5 mL of 1:1:1 methanol:acetonitirle:acetone |
| Extract Storage: | 4℃ |
| Sample Resuspension: | 9:1 Water:Methanol for Reversed phase - CSH; 85:15 acetonitrile:water for HILIC |
