Refer to A-20, p.21, p.26, p.27
As the filler of these columns, a styrene-divinylbenzene-polymethacrylate resin optimized for pretreatment is used. This filler has the advantages that the retention of polar compounds is higher and the adsorption of basic compounds owing to interaction is lower compared to silica-based fillers. It can be applied in a wide range of fields, including analysis of chemical drugs in biological samples, such as serum, and environment samples.


Column:Presep®-C RPP(Short)
Column conditioning:① CH3CN 5 mL
              ② CH3OH 5 mL
              ③ H2O  10 mL
   ↓
* Water sample: 1000 mL
   ↓
Cleaning : 5% CH3OH 3 mL
   ↓
Drying : Suction of air for 5 min
   ↓
Elution : CH3CN 3 mL
* The water sample is prepared with 1 mol/L HCl
   to obtain pH 3.5.
   River water is filtered to remove turbidity.

Column :Wakopak® Navi C18-5 (4.6 mm × 150 mm)
Eluent :CH3CN/50 mM NaClO4 (pH 2.6← H3PO4)=34/66 (v/v)
Flow Rate :1.0 mL/min. at 35℃
Detection :ECD 900 mV, Range= 1, ATTEN= 5
Injection Volume:10 μL
Sample :1)Estriol (E3), 2)17β-estradiol (E2),
      3)Ethynylestradiol (EE2),4)Estron(E1)


Sample water 1000 mL of purified water 1000 mL of river water
Amount of added reference standard 1 μg 0.3 μg 0.1 μg 1 μg 0.3 μg 0.1 μg
Estriol (E3) 98.3 89.9 76.2 - * - * - *
17β-estradiol (E2) 96.6 97.5 90.6 97.9 94.5 91.2
Ethynylestradiol (EE2) 97.9 94.7 86.0 99.9 95.8 88.8
Estron(E1) 101.6 91.7 80.6 105.2 103.4 91.5
* The quantity of estriol could not be determined because it was contained in the river water.


Column:Presep® RPP (60 mg/3 mL)
Column conditioning:① CH3CN    3 mL
              ② Phosphate buffer 3 mL
   ↓
Serum sample: 1 mL of serum and 1 mL of phosphate buffer are mixed and fed into the column (5 μ g of drug reference standard is added).
   ↓
Cleaning : 1 mL of phosphate buffer
   ↓
Dehydration : Air or N2 gas for 5 min
   ↓
Elution : CH3OH 1 mL
   ↓
Injection of 10 μ L into HPLC

* Phosphate buffer: 20 mM phosphate buffer (pH 7)
* Serum: Fetal bovine serum

Column :Wakopak® Wakosil-Ⅱ 5C18 RS (4.6 mm × 150 mm)
Eluent :CH3CN/20 mM Na2HPO4, NaH2PO4 (pH 7.6)=13/87 (v/v)
Flow Rate :1.0 mL/min. at 40℃
Detection :UV 254 nm
Injection Volume:10 μL
Standard :5 μg/mL (CH3OH)
Sample :1)Procainamide HCI, 2)N-Acetyl Procainamide HCl

■ Recovery test
Recovery (%)
n=4
Procainamide N-Acetyl Procainamide
Average
recovery
(%)
CV(%) Average
recovery
(%)
CV(%)
1 Standard solution 101.6 2.3 97.6 2.9
2 Addition to bovine serum 96.1 4.3 95.0 2.1




← previous page ⇒ to Catalog Index
⇒ to Application Index by Sample
→ next page