BS EN ISO 15302 pdf download

BS EN ISO 15302 pdf download Animal and vegetable fats and oils — Determination of benzo[a]pyrene content — Reverse-phase high-performance liquid chromatography method
1 Scope
This lnternational Standard specifies a method for the determination of benzolalpyrene in crude or refined edibleoils and fats by reverse-phase high-performance liquid chromatography (HPLC) using fluorimetric detection in therange from 0,1 ug/kg to 10 g/kg.
2 Normative reference
The following standard contains provisions, which through reference in this text,constitute provisions of thisIntermational Standard. At the time of publication the edition indicated was valid. All standards are subject to revision
and parties to agreements based on this lnternational Standard are encouraged to investigate the possibility ofapplying the most recent edition of the standard indicated below. Members of lEC and ISO maintain registers ofcurrently valid International Standards.
SO 661:1989, Animal and vegetable fats and oils — Preparation of test sample
3 Principle
Adsorption of a suitable amount of sample on an alumina column, followed by elution with light petroleum of anybenzolalpyrene present, and subsequent analysis of the eluate by HPLC using a fuorimetric detector.
4Reagents
All reagents shall be of recognized analytical grade. Where analytical grade solvents other than the recommendedones are used, a full blank analysis shall be carried out and the results of this blank analysis reported
4.1 Water, double distilled, filtered through a membrane filter of 0,45 pm pore size; deionized water obtained bypurifying demineralized water systems may also be used.
4.2 Light petroleum (boiling point range between 40 °C and 60 °C), or hexane, redistiled over potassiumhydroxide pellets (4 g/l).
4.3 Acetonitrile, suitable for HPLC
4.4Tetrahydrofuran, suitable for HPLC
4.5 Toluene, suitable for HPLC
4.6Sodium sulfate, granular, anhydrous
.7Alumina, activity grade 4, prepared from neutral aluminium oxide, activity grade super 1,deactivated by theaddition of 10 ml distilled water to 90 g of alumina.
CAUTION -THE REACTION IS EXOTHERMIC AND PRESSURE MAY BUILD UPShake the container for about 15 min and allow the contents to equilibriate for 24 h. Store the alumina in a closedvessel at ambient temperature.
4.8 Benzo[a]pyrene, 99,0 % purity.
CAUTION – BENZOJPYRENE IS A KNOWN CARCINOGEN. CARRY OUT ALL WORK WITH IT IN A FUMEHOOD,WEARING GLOVES TO MINIMIZE EXPOSURE
4.9Benzo[a]pyrene solutions1)
4.9.1 Stock solution
Weigh, to the nearest 0,1 mg, about 12,5 mg of benzolapyrene in a 25 ml graduated flask. Dissolve it in toluene(4.5) and fill to the mark.
This solution contains about 0,5 mg/ml benzolalpyrene and should be stored in the dark at 4 C when it is stable for6 months at least.
4.9.2 Standard solutions
Prepare two standard solutions containing approximately 0,2 pg/ml and 0,01 pg/ml of benzolalpyrene respectively
by diluting aliquots of the stock solution (4.9.1) with acetonitrile (4.3).
5Apparatus
Usual laboratory apparatus and, in particular, the following.
5.1 Glass column for chromatography, 300 mm long, 15 mm internal diameter, fitted with sintered glass discs,and polytetrafluoroethylene (PTFE) tap.
5.2 Water baths (two), maintained at 35 C 1C and 65 C 1C