HyperSep Column Applications

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Doxepin in Rat Serum
Using 60mg 3mL HyperSep Retain PEP Extraction Column (Part Number: 60107-203)
Sample Preparation
Mix 10mL of doxepin aqueous solution (20mg/L) and
30mL of rat serum into a 100mL vessel
Dilute to 100mL with 0.5% ammonia solution to give a
2ppm solution
Condition Retain PEP Extraction Column
2mL of methanol
2mL of DI H2O
Apply Sample
Load 2mL samples
Wash Column
2mL of 0.5% ammonia solution containing 5% methanol
Elute Doxepin
2mL of 1% acetic acid in methanol
Dry Eluate and Reconstitute
Evaporate to full dryness at room temperature under a
stream of nitrogen
Reconstitute the sample to 1mL with
sodium acetate (pH 4) (40:60, v/v)
Analysis
Mobile phase: sodium acetate (pH 4)
(40:60, v/v)
Flow: 1.0mL/minute
Injection: 10µL
Temperature: 30°C
Detection: 290nm
Recommended HPLC Column Part Number
Hypersil GOLD 5µm, 250 x 4.6mm 25005-254630
Drug Analysis in Serum
Using 60mg 3mL HyperSep Retain PEP Extraction Column (Part Number: 60107-203)
Condition HyperSep Retain – PEP Column
1 x 3mL CH3OH
1 x 3mL H2 O
Apply Sample
Load spiked serum sample onto SPE cartridge
Wash Column
1 x 3mL H2 O
1 x 3mL 5% CH 3OH
Elution
1 x 3mL CH3OH
Results
Analytes Recovery (%)
Dexamethasone 97.9
Ethinylestradiol 96.3
Hydrocortisonµm 74.0
Triamcinolone 71.9
Levonorgestrel 93.9
Ganciclovir 54.1
Prednisone Acetate 98.6
Cefalexin 58.6
Cefradine 45.6
Pharmaceutical/Biotech
Applications