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MB Sample ID: SA206131

Local Sample ID:QC_Negative_MS_Pool_Front3
Subject ID:SU002237
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606

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Subject:

Subject ID:SU002237
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
QC_Negative_MS_Pool_Front3SA206131FL025279QCTreatment

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
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