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MB Sample ID: SA255662

Local Sample ID:Toxoplasma infected-6
Subject ID:SU002646
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090

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Subject:

Subject ID:SU002646
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
Toxoplasma infected-6SA255662FL032735InfectedFactor

Collection:

Collection ID:CO002639
Collection Summary:Serum collected from T. gondii-infected mice at 9 wk post-infection. Serum samples from uninfected mice were included as controls.
Sample Type:Blood (serum)

Treatment:

Treatment ID:TR002658
Treatment Summary:10–14-week-old male C57BL/6 mice were infected with 10 Me49 bradyzoite cysts by intraperitoneal infection resuspended in 200 Μl PBS per mouse using a 5G 5/8” tuberculin syringe.

Sample Preparation:

Sampleprep ID:SP002652
Sampleprep Summary:Serum samples were stored at -80C before the mass spectrometry assay.

Combined analysis:

Analysis ID AN004193
Analysis type MS
Chromatography type Reversed phase
Chromatography system Waters Acquity
Column Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um)
MS Type ESI
MS instrument type QTOF
MS instrument name Agilent 6530 QTOF
Ion Mode NEGATIVE
Units peak heights

Chromatography:

Chromatography ID:CH003107
Instrument Name:Waters Acquity
Column Name:Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um)
Column Temperature:65
Flow Gradient:15%-99%
Flow Rate:0.6 ml/min
Solvent A:60% water/40% acetonitrile; 5 mM ammonium formate; 0.1% formic acid
Solvent B:90% isopropanol/10% methanol; 5 mM ammonium formate; 0.1% formic acid
Chromatography Type:Reversed phase

MS:

MS ID:MS003940
Analysis ID:AN004193
Instrument Name:Agilent 6530 QTOF
Instrument Type:QTOF
MS Type:ESI
MS Comments:Samples extracted using Matyash extraction procedure which includes MTBE, MeOH, and H2O. The organic (upper) phase was dried down and submitted for resuspension and injection onto the LC while the aqueous (bottom) phase was dried down and submitted to derivatization for GC. They are resuspended with 110 uL of a solution of 9:1 methanol: toluene and 50 ng/mL CUDA. This is then shaken for 20 seconds, sonicated for 5 minutes at room temperature, and then centrifuged for 2 minutes at 16100 rcf. The samples are then aliquoted into three parts. 33 uL are aliquoted into a vial with a 50 uL glass insert for positive and negative mode lipidomics. The last part is aliquoted into an eppendorf tube to be used as a pool. The samples are then loaded up on an Agilent 1290 Infinity LC stack. The positive and negative mode were run on an Agilent 6530 with a scan range of m/z 120-1200 Da with an acquisition speed of 2 spectra/s. Between 0.5 and 2 uL were injected onto a Waters ACQUITY UPLC CSH C18 1.7um 2.1x100mm Column with VanGuard PreColumn 2.1x5 mm . The gradient used is 0 min 15% (B), 0–2 min 30% (B), 2–2.5 min 48% (B), 2.5–11 min 82% (B), 11–11.5 min 99% (B), 11.5–12 min 99% (B), 12–12.1 min 15% (B), 12.1–15 min 15% (B) with a flow rate of 0.6 mL/min. The mass resolution for the Agilent 6530 is 10,000 for ESI (+).
Ion Mode:NEGATIVE
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