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Updated: May 23, 2026

High Content Screening Analysis to Evaluate the Toxicological Effects of Harmful and Potentially Harmful Constituents (HPHC)
Published on: May 10, 2016
[Chemical-toxicological determination of 4-methoxyhydroxybenzene]
A P Chernova1, V K Shormanov2, M K Elizarova3
1National Research Tomsk Polytechnic University, Tomsk, Russia.
Objective:
To investigate the features of the chemical and toxicological determination of 4-methoxyhydroxybenzene (4-MOHOB).
Material And Methods:
The methods of the study were TLC, reversed-phase column chromatography of ordinary pressure, GC-MS and HPLC. 4-MOHOB was isolated from liver tissue and blood by infusion with a mixture of ethyl «acetate-acetone» (7:3).
Results:
The expediency of eluting the analyte from the Silasorb C18 column (150×10 mm) with the mobile phase of «acetonitrile-water» (4:6) was shown, which in combination with extraction formed the basis for the developed version of purification of 4-MOHOB extracted from biomatrices. The advantages of the «hexane-ether» (6:4) mobile phase in the identification of 4-MOHOB by TLC (Merck plates) were proven. One of the variants of 4-MOGOB identification is based on its extinction properties (UV range; medium - 95% ethanol). GC-MS (mobile phase - helium, stationary phase «5% phenyl-95% methylpolysiloxane» in a column measuring 25 m × 0.2 mm) was used as the main method of confirmatory identification of 4-MOGOB. The mass spectrum of this compound included signals of the following ions (m/z): 38, 53, 65, 81, 109, 124. A 250×4.6 mm Discovery C 18 column and an eluent of «acetonitrile-acid-water solution with a pH of 5.5» (50:50) were proposed for determining 4-MOGOB by HPLC.
Conclusion:
The compliance of the developed methods with the key validation parameters was confirmed. The detection limits of 4-MOHOB in 100 g of organ (liver) and blood are 0.08 and 0.06 mg, respectively, the limits of quantitative determination are 0.14 and 0.10 mg.
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