Review Team (February 2016).pdf

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Methods
(Applicable for determination of folic acid and 5Ͳmethyltetrahydrofolate in reconstituted
adult/infant nutritional formula)
A. Principle
Folate extraction involved treatment in a boiling water bath, solid phase extraction and
ultraͲfiltration. Quantitation of folic acid and 5Ͳmethyltetrahysdrofolate is done by ultraͲ
performance liquid chromatographyͲtandem mass spectrometry (UPLCͲMS/MS).
B. Apparatus
(a) Analytical balance
ƒ XS 105 Mettler Toledo, Port Melbourne, Australia
ƒ Kern AEJ 220Ͳ4M, Balingen, Germany
(b) Water bath Julabo TW 20, Seelbach, Germany
(c) pH meter, TPS digital, Brisbane, Australia
(d) Vortex mixer, Ratex, Boronia, Australia
(e) Beakers (200 mL)
(f) Plastic spoons
(g) Magnetic stirrer
(h) Electric water kettle, Sunbeam, Sydney, Australia
(i) Speed vacuum, Savant Thermo Scientific, Sydney, Australia
(j) Centrifuge/falcon tubes (50 mL)
(k) Measuring cylinders (100 and 250 mL)
(l) Volumetric flasks (5, 10, 50 mL)
(m)Eppendorf tubes (1.5 mL)
(n) Automatic pipettes, Gilson, Sydney, Australia
(o) eVol® XR handͲheld automated analytical syringe, SGE Analytical Science, Melrose
Park, Australia
(p) Centrifuge, Heraeus Multifuge X3R, Thermo Fisher Scientific, Sydney, Australia
(q) Centrifuge, Heraeus Biofuge, Germany
(r) Amicon UltraͲ2, UltracelͲ3 Membrane, 3 kDa cutͲoff, Millipore, Bedford, MA
(s) Filter paper Whatman 41 ashͲless
Fol-20 (February 2016)
FOR ERP USE ONLY
DO NOT DISTRIBUTE