Test

Method, analyser, reagents

Laboratory/reference

Neurotransmitters

Biogenic amines: dopamine, noradrenaline and adrenaline,

Spot-baseline (fasting) urinary neurotransmitter testing (second void morning), snap-frozen to minus 30 degrees and analysed by mass spectrometry, using nanomols per millimol of urinary creatinine as a standard.

SA Pathology, Adelaide, South Australia.

Whiting MJ. 2009. Simultaneous measurement of urine metanephrines and catecholamines by liquid chromatography with tandem mass spectrometric detection. Annals of Clinical Biochemistry, 46, 129-136.

Creatinine

Spot urine specimen from the same void as biogenic amines, expressed in millimols per Litre.

SA Pathology, Adelaide SA.

Oxidative stress: Urinary hydroxyhemopyrroline-2-one

Urinary hydroxyhemopyrroline-2-one (HPL)

Spot urine (second void morning) in ascorbic acid, snap frozen (-30C) and light-protected. Colorimetric method at 540nm, following solvent extraction and reaction with Erich’s reagent. micrograms per decilitre.

Applied Analytical Laboratories,

8/26 Nestor Drive, Meadowbrook, Queensland, Australia. +61 7 3133 1615.

Nutrition-Biochemistry

Vitamin D (25-OH)

Diasorin Liason assay kit. for use on the Liaison platform nmol/L.

Clinpath Laboratories, 19 Fullarton Rd, Kent Town. South Australia 5067+61 8 8366 2000.

Serum total Vitamin B12

Competitive Electrochemiluminescent Immunoassay. Roche Modular E 170.

Automated Immunoassay Analyser,

using Roche Vitamin B12 Reagent (pmol/L).

Clinpath Laboratories. As above.

Red Cell Folate

Competitive Electrochemiluminescent Protein Binding Assay, using Roche Modular E 170, using Roche Folate Red Blood Cell (RBC) Reagent and Roche Folate RBC Haemolysing Reagent. on Automated Immunoassay Analyser (nmol/L).

Clinpath Laboratories. As above.

Vitamin B6 (Pyridoxal-5’-phosphate coenzyme form)

Whole blood High Pressure Liquid Chromatography with fluorescent detection. Chromsystems Vitamin B6 in Whole Blood High pressure Liqid Chromatography Reagent Kit. Waters Alliance 2695 Separations Module. Waters 474 Fluorescence Detector (nmol/L).

Sullivan Nicolaides Pathology

143 Whitmore St, Taringa. Queensland 4068. Australia. +61 7 337 8666

Serum Copper

Flame Atomic Absorption Spectrophotometry.

Varian AA-240FS (umol/L).

Douglass Hanly Moir Pathology

14 Griffnock Avenue, Macquarie Park.

New South Wales 2113. +61 2 9855 5222.

Red Cell Zinc

Inductively coupled plasma mass spectroscopy (ICP-MS), using 6% n-Butanol reagent and Agileny ICP-MS 7500ce analyser (umol/L).

Sullivan Nicolaides Pathology.

As above.

Serum Ceruloplasmin

Immunoturbidimetric method, using 6K91-30

Multignet Caeruloplamin Kit and Abbott Architect ci16000 analyser (g/L).

Douglass Hanly Moir Pathology.

As above.

Percentage Free Copper/Red Cell Zinc

Percentage of free copper in the serum calculated by an equation based on the molecular and atomic weights of ceruloplasmin and copper (one ceruloplasmin molecule binds to six copper atoms). The ratio of the percentage free copper to red cell zinc was calculated as “percentage free copper”/“Red cell zinc umol/L”.

Calculated by authors

Intermediate substrates

and enzymes

MTHFR Ala222Val (C677T) methyl tetrahydrofolate reductase polymorphism

Real time PCR analysis

Roche Diagnostics Light-Cycler 480 kit. Using

TecnoBiol reagents, Sigma probes and primers on Roche LC480 analyser.

Douglass Hanly Moir Pathology.

As above.

Plasma homocysteine

Ice transported EDTA sample. Competitive Chemiluminescent Immunoassay, using Seimens Homocysteine reagent on Seimens Advia centaur Automated Immunoassay (umol/L).

SA Pathology.

+61 8 8222 3000

As above.

Serum histamine

Beckman Coulter Radio Immunoassay, using Beckman Coulter R.I.A. Kit on Perkin Elmer Wizard 1470 Automated Gamma Counter (umol/L).

Sullivan and Nicolaides.

As above.