EXtrelut® NT |
Application Example
EXtrelut® NT columns for liquid-liquid extraction have been used extensively for the sample preparation of urine, whole blood, plasma, serum, gastric juice, liquor, amniotic fluid, and faeces, as well as animal and plant tissues. Other suitable application fields are environmental and residue analyses, including the analysis of industrial, domestic and waste water. The fractionated elution of acidic and basic substances, such as drugs and their metabolites from body fluids, is also possible with EXtrelut® NT columns.
Determination of Antiepileptic Drugs (AEDs) in Serum
| 500 µL serum 500 µL phosphate buffer* |
|
| ↓ | Apply in sequence onto the column |
| EXtrelut® NT1 | |
| ↓ | Wait 8 minutes |
| 1 mL dichloromethane / 2-propanol (9+1) |
|
| ↓ | Wait 10 minutes then elute with |
| 6 mL dichloromethane / 2-propanol (9+1) |
|
| ↓ | Evaporate to dryness under nitrogen stream |
| Redissolve residue in 1 mL of methanol | |
| ↓ | |
| Inject 10 µL onto HPLC column |
* = 17.6 g NaH2PO4, 4.5 g Na2HPO4*2H2O, 1.5 g NaN3, dissolved in 1 L water (pH 6.0-6.1)
HPLC Separation of AEDs After Sample Preparation with EXtrelut® NT1
| HPLC System | LaChrom® System | ||||||||||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Column | LiChrospher® RP-select B (5µm) LiChroCART® 250-4 |
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| Mobile phase | A: Water LiChrosolv®, Acetonitrile LiChrosolv® / 1+1 (1.00030) B: Water LiChrosolv® |
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| Gradient |
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| Flow | 1 mL/min | ||||||||||||||||||||||||||
| Temperature | 30°C | ||||||||||||||||||||||||||
| Detection | UV 205 nm | ||||||||||||||||||||||||||
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Recoveries (mean values N=3)
| 1 Ethosuximide* | 14.1 min | 84 ± 7% | |
|---|---|---|---|
| 2 Primidone | 19.4 min | 100 ± 2% | |
| 3 a-Methyl-a-propylsuccinimide | 22.5 min | Internal standard | |
| 4 Phenobarbital | 26.9 min | 96 ± 2% | |
| 5 Hexobarbital | 34.2 min | 99 ± 2% | |
| 6 Carbamazepine | 36.4 min | 97 ± 1% | |
| 7 Phenytoin | 38.0 min | 100 ± 1% | |
*Ethosuximide is volatile on evaporation
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