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MB Sample ID: SA206226
Local Sample ID: | Q5_Positive_2 |
Subject ID: | SU002237 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU002237 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
Q5_Positive_2 | SA206226 | FL025281 | preeclampsia | Treatment |
Collection:
Collection ID: | CO002230 |
Collection Summary: | Placentas were first processed by collecting samples ~4-6 inches in length and ½ inch in diameter via cross-section punch biopsy, with blinded identifiers to ensure unbiased collection. These samples were stored at -80°C, until further processing on dry ice into individual samples for chemical analyses. Here, sterile scalpels were used to cut samples in half and a ¼ inch slice was removed from the middle and weighed, yielding isolated samples ranging in weight between 0.4 and 0.6 g. |
Sample Type: | Placenta |
Treatment:
Treatment ID: | TR002249 |
Treatment Summary: | Placentas were first processed by collecting samples ~4-6 inches in length and ½ inch in diameter via cross-section punch biopsy, with blinded identifiers to ensure unbiased collection. These samples were stored at -80°C, until further processing on dry ice into individual samples for chemical analyses. Here, sterile scalpels were used to cut samples in half and a ¼ inch slice was removed from the middle and weighed, yielding isolated samples ranging in weight between 0.4 and 0.6 g. |
Sample Preparation:
Sampleprep ID: | SP002243 |
Sampleprep Summary: | One sample per tissue was used in chemical analyses. Chemicals were separated from tissue via solid-liquid extraction by adding 2 mL of room temperature water:acetonitrile (1:1) per sample. Placenta samples were disrupted and homogenized using a TissueRuptor (Qiagen), vortexed for 5 min, and placed in a centrifuge at 4000 rpm for 5 min. The resulting supernatant was collected, and a second round of solid-liquid extraction was performed using 8 mL acetonitrile with 2% formic acid. Samples were vortexed and centrifuged again, and both supernatants combined yielding a final total volume of 10 mL. Of the 10 mL combined supernatant sample, 3 mL were then run through solid filtration with a Captiva EMR-lipid column (Agilent Technologies). Resulting eluates were placed into liquid chromatography mass spectrometry (LCMS) vials in 1.0 mL aliquots. Remaining supernatant volumes were banked at -80°C. Method blanks were collected in parallel using the same steps without adding tissues. For non-targeted chemical analysis, 20 µL of 1.25 µg/mL tracer solution was added to each 1.0 mL aliquot of post-filtration eluate (for study samples, blanks, and controls) yielding a final concentration of 25 ng/mL per sample. In addition, tracers were added to a single sample in duplicate prior to extraction to evaluate extraction recovery and reproducibility. |
Combined analysis:
Analysis ID | AN003522 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Agilent 1290 Infinity II |
Column | Agilent Zorbax Eclipse Plus C8 (150 x 2.1mm,1.8 um) |
MS Type | ESI |
MS instrument type | QTOF |
MS instrument name | Agilent 6545 QTOF |
Ion Mode | UNSPECIFIED |
Units | amu |
Chromatography:
Chromatography ID: | CH002601 |
Instrument Name: | Agilent 1290 Infinity II |
Column Name: | Agilent Zorbax Eclipse Plus C8 (150 x 2.1mm,1.8 um) |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS003280 |
Analysis ID: | AN003522 |
Instrument Name: | Agilent 6545 QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | See Acquisition_Methods.docx for more comprehensive acquisition parameters |
Ion Mode: | UNSPECIFIED |