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Last updated date: March 30, 2015
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Reports

Advisory Committee on Health Effects of Endocrine Disruptors
The Supplement II to the Intermediary Report
1.2.3.2

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Part 2
Analysis of bisphenol A in biological samples

[Outline]
(i) LC/MS1: Serum or urine samples cleaned up with a multimode cartridge are analyzed by a high performance liquid chromatograph/mass spectrometer (LC/MS) qualitatively and quantitatively.
(ii) GC/MS2: Urine samples, cleaned up with a C18 cartridge in acid atmosphere, trimethylsilylated, and further cleaned up with a Florisil cartridge, are analyzed by a gas chromatograph/mass spectrometer (GC/MS) qualitatively and quantitatively.

[Reagents]
Confirm by chromatography that all the reagents present no problem in bisphenol A (BPA) analysis.
(i) Standard samples: Environmental analysis grade BPA and BPA-d16, and 13C-BPA supplied by Cambridge Isotope.
(ii) Standard BPA solution: Place 100 mg standard BPA in a 100-ml measuring flask, and dissolve by adding methanol to obtain a concentrated standard solution of 1000ƒĘg/ml. Prepare appropriate standard solutions by diluting the concentrated standard solution.
(iii) Internal standard solution: Place 10 mg BPA-d16 (for LC/MS) or 18C-BPA3 (for GC/MS) in a 50-ml measuring flask, and dissolve by adding methanol to obtain a concentrated standard solution of 2000 ƒĘg/ml. Add the internal standard solution to the standard and sample solutions to a temporary concentration and analyze by LC/MS or GC/MS.
(iv) ƒŔ-Glucuronidase: A solution of at least 100,000 units/ml.
(v) Cartridges for purification: A multimode cartridge (500 mg)4 and a C18 cartridge (500 mg)5. Wash the cartridge with 10 ml methanol and then 5 ml water before use. Wash the Florisil cartridge (500 mg)6 with 5 ml n-hexane before use.
(vi) BSTFA7: Environmental analysis grade.
(vii) Pure water: Milli-Q water hardly contaminated by BPA.
Other reagents should be of the special grade or HPLC grade8.

[Tools]
Test the tools for sampling and sample preparation8 beforehand for material or leaching, and choose ones with as little BPA release as possible. Heat glassware to 350Ž for at least two hours, let cool down in a place free from BPA contamination from the environment, and wash with acetone immediately before use.

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