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Biogenic Amines by LCMS using Kinetex 1.7u C18 50x3.0mm

Sample Preparation Details

Part No:8B-S029-UBJ
Solid Phase Extraction (SPE) procedure:Note: The solvent volumes shown below are for a 60 mg bed mass. The solvent volumes will need to be adjusted for a smaller or larger bed mass.
Load:Salmon samples were pre-treated prior to loading onto the conditioned Strata-X-C sorbent:
1. weigh out 4.0 g of salmon into a 50 mL plastic centrifuge tube
2. pulverize meat using a spatula against the sides of the centrifuge tube
3. add 20 mL Acetonitrile and 12 mL of Hexane, vortex for 1 minute
4. centrifuge for 5 minutes at 14,500 rpm
5. transfer the bottom Acetonitrile layer to a new tube
6. repeat extraction with a new 20 mL aliquot of Acetonitrile, vortex for 1 minute then centrifuge
7. collect the lower layer and combine fractions

The combined fractions are now ready to loaded onto the conditioned Strata-X-C sorbent.
Final Prep and Analysis: Inject: 2 µL on HPLC / Tandem Mass Spec (MS-MS) @ (ambient)

Analytes:

Name Spiked
Conc.
Log P pKa % Rec %RSC (n=0)
1 IQx
2 8-MeIQx
3 IQ
4 7,8-DiMeIQx
5 4,8-DiMeI-Qx
6 MeIQ
7 Trp-P-2
8 Harman
9 PhIP
10 Norharman
11 AaC
12 MeAaC

Application Detail (App ID: 20685)

Part No:00B-4475-Y0
Dimensions:50 x 3 mm ID
Elution Type:Gradient
Elution A:30mM Ammonium formate
Elution B:Acetonitrile
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 0 80 20
2 3 40 60
3 3.1 0 100
4 4 0 100
5 4.1 80 20
6 5 80 20

Flow Rate:600 µL/min
Col. Temp: ambient
Detection: Tandem Mass Spec (MS-MS) @ (22°C)
Detector Info: AB Sciex API 4000™

Analyte Details

Analyte Detail not available

1
IQx
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2
8-MeIQx
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3
IQ
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4
7,8-DiMeIQx
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5
4,8-DiMeI-Qx
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6
MeIQ
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7
Trp-P-2
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8
Harman
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9
PhIP
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10
Norharman
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11
AaC
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12
MeAaC