Sterols from Margarine on ZB-5MS

Sample Preparation Details

Part No:8B-S001-JCH
Solid Phase Extraction (SPE) procedure:Note: The solvent volumes shown below are for a 1 g bed mass. The solvent volumes will need to be adjusted for a smaller or larger bed mass.
Load:0.72 g Margarine in in 20 ml 0.5 M KOH in ethanol. Shake and reflux at 78 °C for 20 min. Cool down then add 10m 0.22 mg/ml betulin (istd) and shake for 5 min. Let it still to settle then collect liquid phase into a 20 ml screw cap vial. Pipette and filter 3ml aliquot through
a BD 10 ml syringe screw to Phenex 30 mm 0.45 um Teflon filter and collect it in a 4 ml screw cap vial. Set up and condition Strata C 18-E SPE (55
um, 70 A) on a manifold to vacuum pump onto a 1/125 mm culture tube, with 5 ml methanol and 5 ml chloroform.
Replace collected conditioning material with a new tube and label accordingly. Pipette 1 ml sample filtrate onto chloroform-SPE sorbent miniscuos and elute it with 15 ml 5 % methanol in chloroform. Concentrate eluted sterols to 1 ml by drying it with nitrogen. Transfer 200 ul aliquot to 500 ul insert sample vial and dry it with nitrogen then reconstitute with 100 pyridine and derivatize with 100 ul BSTFA:TMCS; 99:1. Warm up at 70 °C for 20 min and inject.

and label.
Final Prep and Analysis: Initial Sample Unit: gram Inject: 0.5 µL on GC / Flame Ionization (FID) @ / (320 °C)


Name Spiked
Log P pKa % Rec %RSC (n=0)
1 Campesterol
2 Stigmasterol
3 b-Sitosterol
4 Betulin

Application Detail (App ID: 16512)

Part No:7HG-G010-11
Phase:5% Phenyl-Arylene 95% Dimethylpolysiloxane
Dimensions:30 meters x 0.25 mm x 0.25 µm
Oven Profile:225 °C to 330 °C @ 15 °C/min for 2 min.
Carrier Gas:Constant Flow Helium, 2.5 mL/min
Injection:Splitless :1 0.5 µL @ 320°C
Detection: Flame Ionization (FID) (320°C)
Note:Sample prep- Saponification, Solid Phase Extraction, and Derivatization.

Analyte Details

Analyte Detail not available