HyperSep Column Applications

Benzodiazepines in Whole Blood for GC or GC/MS Confirmations

Using 100mg 1mL HyperSep Diol Extraction Column

(Part Number: 60108-572)

Prepare Sample

To 1mL of pH 6 buffer add internal standards*, add 2mL

of whole blood and mix/vortex

Add 5mL of pH 6 buffer

Sonicate with a probe sonifier for ~10 seconds and

centrifuge at ~2700rpm for 15 minutes

Condition HyperSep Diol Extraction Column

1 x 3mL ethyl acetate

1 x 3mL MeOH

1 x 3mL DI H

1 x 3mL 0.1M phosphate buffer (pH 6.0)

Apply Sample

Load sample by gravity

Wash Sample

1 x 3mL DI H

1 x 3mL 5% acetonitrile in 0.1M phosphate

buffer (pH 6.0)

Dry columns 5 minutes at full vacuum or >10"Hg

1 x 3mL hexane

Elute Benzodiazepines

2 x 3mL ethyl acetate

Dry Eluate

Evaporate to dryness under nitrogen at ~55°C

Add external standards*

Derivatize

Add 100µL acetonitrile and 100µL MTBSTFA

w/1% t-BDMCS

Heat for 30 minutes at 70°C

Remove from heat source to cool

Inject 1µL into GC/MS-NCI

NOTE:

Do not evaporate MTBSTFA solution

* Suggested internal standards: Diazepam-D5 and Lorazepam-D4.

Recommended GC Column

Part Number

TraceGOLD TG-5MS 30m x 0.25m x 0.25µm

26098-1420

Pharmaceutical/Biotech

Applications