The use of both Presep®-C RPP (Short) and Presep®-CM for pretreatment can improve the cleanup effect.



Column :Presep®-C RPP(Short)
Conditioning:① CH3OH 10 mL
       ② H2O 10 mL
       ③ Saturated EDTA・2Na, 5 mL
         Sample *1
          ↓
         Cleaning:H2O 10 mL
          ↓
         Elution *2:CH3OH 5 mL
          ↓
Column :Presep®CM
Conditioning:① CH3OH 10 mL
         ② 0.1mol/L HCl 10 mL
         ③ H2O 20 mL
         ④ CH3OH 5 mL
          ↓
    Cleaning:CH3OH 5 mL
          ↓
    Elution:CH3CN/0.1mol/L HCl=1/1(v/v) 5 mL
          ↓
    Condensation to 2 mL or less under flow of N2 gas
          ↓
    Diluted to 2 mL with H2O
          ↓
    Pouring of 30 μL of HPLC




Column :Wakopak®Navi C18-5
      (4.6 mm×150 mm)
Eluent :CH3OH/Imidazole
     Buffer=25/75(v/v)
Flow Rate:1.0 mL/min. at 40℃
Detection:FLD ex. 380 nm, em. 520 nm
Sample :1.Oxytetracycline(OTC)
      2.Tetracycline(TC)
      3.Cholortetracycline(CTC)


■Standard solution:Presep®-C RPP(Short)
  OTC TC CTC
1 99.6 97.2 96.2
2 102.1 100.8 98.5
3 105.2 102.3 100.0
4 100.8 99.0 97.5
101.9 99.8 98.0
CV(%) 2.4 2.2 1.6
*Standard solution sample: 1 μg of each reference standard is added to 50 mL of citric acid buffer containing EDTA.

■Standard solution:Presep®CM
  OTC TC CTC
1 96.2 89.3 91.2
2 95.1 91.3 94.0
3 97.9 89.9 93.3
4 97.8 91.5 94.5
96.7 90.5 93.2
CV(%) 1.4 1.2 1.5
*Standard solution sample: 1 μg of each reference standard is added to 5 mL of CH3OH.

■Shrimp extract
  OTC TC CTC
1 96.9 86.8 86.7
2 97.1 86.9 85.7
3 94.8 89.8 88.3
4 95.7 89.9 90.7
96.1 88.3 87.9
CV(%) 1.2 2.0 2.5
*1 μg of each reference standard is added to the shrimp extract sample.

(1) 15 μL of standard drugs added to shrimp extract are treated with Precep
®-C RPP (Short).
(2) 30 μL of standard drugs added to shrimp extract are treated with Precep®-C RPP (Short) and Precep® CM.


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