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MB Sample ID: SA212444

Local Sample ID:37899_Sk
Subject ID:SU002315
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Genotype Strain:C57BL6 and FVB mixed strain
Age Or Age Range:54-59 weeks old
Gender:Male and female
Animal Feed:Specialty Feeds Irradiated Rat and Mouse Standard Chow Diet

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Combined analysis:

Analysis ID AN003638
Analysis type MS
Chromatography type Reversed phase
Chromatography system Agilent 1290 Infinity II
Column Agilent Zorbax Eclipse Plus C18 (100 x 2.1mm, 1.8 um)
MS Type ESI
MS instrument type Triple quadrupole
MS instrument name Agilent 6490 QQQ
Ion Mode POSITIVE
Units pmol per mg (tissues) pmol per ml (plasma)

Chromatography:

Chromatography ID:CH002693
Chromatography Summary:The running solvent consisted of solvent A: 50% H2O / 30% acetonitrile / 20% isopropanol (v/v/v) containing 10mM ammonium formate and 5uM medronic acid, and solvent B: 1% H2O / 9% acetonitrile / 90% isopropanol (v/v/v) containing 10mM ammonium formate. We utilized a stepped linear gradient with a 16-minute cycle time per sample and a 1µL sample injection. To increase throughput, we used a dual column set up to equilibrate the second column while the first is running a sample. The sample analytical gradient was as follows: starting with a flow rate of 0.4mL/minute at 15% B and increasing to 50% B over 2.5 minutes, then to 57% over 0.1 minutes, to 70% over 6.4 minutes, to 93% over 0.1 minute, to 96% over 1.9 minutes and finally to 100% over 0.1 minute. The solvent was then held at 100% B for 0.9 minutes (total 12.0 minutes). Equilibration was started as follows: solvent was decreased from 100% B to 15% B over 0.2 minutes and held until a total of 16 minutes. The next sample is injected and the columns are switched.
Instrument Name:Agilent 1290 Infinity II
Column Name:Agilent Zorbax Eclipse Plus C18 (100 x 2.1mm, 1.8 um)
Flow Gradient:starting with a flow rate of 0.4mL/minute at 15% B and increasing to 50% B over 2.5 minutes, then to 57% over 0.1 minutes, to 70% over 6.4 minutes, to 93% over 0.1 minute, to 96% over 1.9 minutes and finally to 100% over 0.1 minute. The solvent was then held at 100% B for 0.9 minutes (total 12.0 minutes). Equilibration was started as follows: solvent was decreased from 100% B to 15% B over 0.2 minutes and held until a total of 16 minutes.
Flow Rate:0.4mL/min
Solvent A:50% water/30% acetonitrile/20% isopropanol; 10mM ammonium formate; 5uM medronic acid
Solvent B:1% water/9% acetonitrile/90% isopropanol; 10mM ammonium formate
Chromatography Type:Reversed phase
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