Step 1
AccQFluor Reagent 2A(2A Solution)Preparation
Preheat the oven or the dry block heater to 55 ℃.
Open 2B reagent(high purity Acetonitrile with out waters)carefully.
Open 2A reagent and add 1 mL 2B, votex for at least 30 s, until 2A powder desolved completely.
Screw the cap of 2A bottle and incubate at 55 ℃ for 10 min.
Tips
The derivatization soluton (2A solution) should be kept water-free completely.
The preperaed 2A solution could be separated for the suitble volume into the completely sealed glass bottle, tighten the cap by sealing film, and stored into the dryer for standby.
Step 2
Sample Preparation
Plasma/Serum
10 μL plasma/serum +30 μL ACN in a 1.5 mL eppendorf tube.
Vortex for at least 10 s.
Centrifuge at 4 ℃, 12000 rpm, for 10 min.
Tanslate 10 μL supernatant carefully to the Sample Tubes (WAT007571,72pkg) for the next derivatization (Step 3)
Step 3
Derivatization
Add 70 μL AccQFluor Reagent 1 (in kit WAT052875) into Sample Tubes(WAT007571, 72pkg) which contains 10 μL supernatant (Step 2).
Vortex 10 s for completely mixing.
Continue vortex and entirely inject 20 μL 2A solution (preparaed in Step 1) into the vortexing Sample Tubes, as quickly as possible!
Keep on Votex for 10 s after adding 2A solution.
Translate all the mixture into a vial, and screw the cap.
Incubat the vial at 55 ℃ for 10 min.
Tips
Step 4
The derived Sample could keep stable in 4 weeks, sealed at 4 ℃.