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Derivatization of Amino Acid by Waters AccQ-Tag Kit
Author: Li Quan, updated date: , view: 31, Q&A: 0

Materials and Reagents

  1. Amino Acid Analysis (AAA) is required for many applications. 
    Refill Package. The refill Kit is intended to recharge the AccQTag Ultra chemistries that are a part of the Application Add on kit. This kit should not be purchased as part of an initial system. 
    Includes AccQTag Ultra Deriviativation Kit 250 analyses 186003836, 
    AccQTag Ultra Column 2.1 × 100mm 186003837, 
    AccQTag Eluent A Concentrate (950mL) 186003838, 
    AccQTag Ultra Eluent B (950mL) 186003839, 
    Sample Tubes, 4 × 72pkg WAT007571, 
    Amino Acid Standard, Hydrolyste, 10 × 1mL ampules WAT088122, 
    Tota Recovery Vials, 3 × 100 vial/pkg 186000384C

    产品编号: 176001235

    AccQ-Tag Chemistry Kit (250 Analyses) [WAT052875]

    Amino Acid Analysis (AAA) is required for many applications.

    ACCQ-Tag Chemistry Kit for up to 250 Analyses includes: 
    AccQFluor Reagent 1 5 × 6mL vials, 
    AccQFluor Reagent 2A 5 × 3mg vials, 
    AccQFluor Reagent 2B 5 × 3 mL vials, 
    AccQTag Column 3.9 × 150mm, 
    AccQTag Eluent A Concentrate 2 × 1L, 
    Sample tubes 4 × 72/pkg, AminoAcid Standard, 
    Hydrolysate 10 × 1mL Ampules WAT088122, 
    AccQTag User Guide WAT052874
     

  2. Reagent

    Methanol

    Acetonitrile

  3. ##### Standard

    Amino Acid


Equipment

  1. Vortex

  2. Oven or dry block heater


Procedure

Step 1
AccQFluor Reagent 2A(2A Solution)Preparation 

  1. Preheat the oven or the dry block heater to 55 .

  2. Open 2B reagent(high purity Acetonitrile with out waters)carefully.

  3. Open 2A reagent and add 1 mL 2B, votex for at least 30 s, until 2A powder desolved completely.

  4. 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

  1. 10 μL plasma/serum +30 μL ACN in a 1.5 mL eppendorf tube.

  2. Vortex for at least 10 s.

  3. Centrifuge at 4 , 12000 rpm, for 10 min.

  4. Tanslate 10 μL supernatant carefully to the Sample Tubes (WAT007571,72pkg) for the next derivatization (Step 3)

Step 3
Derivatization

  1. Add 70 μL AccQFluor Reagent 1 (in kit WAT052875) into Sample Tubes(WAT007571, 72pkg) which contains 10 μL supernatant (Step 2).

  2. Vortex 10 s for completely mixing.

  3. Continue vortex and entirely inject 20 μL 2A solution (preparaed in Step 1) into the vortexing Sample Tubes, as quickly as possible!

  4. Keep on Votex for 10 s after adding 2A solution.

  5. Translate all the mixture into a vial, and screw the cap.

  6. Incubat the vial at 55 for 10 min.

Tips

  • The 2A solution could react with waters and become invalid in seconds. So the 3 substeps must be operate as carefully, precisely and quickly as you can.

Step 4

The derived Sample could keep stable in 4 weeks, sealed at 4 .


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