Determination of residual amounts of beta-adrenostimulants in muscle tissue and liver by dispersion purification chromatography-mass spectrometry

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Abstract

A selective technique for the determination of 20 beta-adrenostimulants in muscle tissue and liver by chromatography-mass spectrometry has been developed. The limit of quantitative determination of the technique is 0.1-0.25 (0.5) µg/kg (depending on the compound to be determined). The technique is based on hydrolysis of research objects followed by extraction of the determined compounds with acetic acid acetonitrile, obtaining the organic fraction by salting, its purification on aluminum oxide layer and dispersive purification with C18, further purification by liquid-liquid extraction with hexane. Data on promising ion-transitions of some determined compounds are given; detection parameters of bambuterol, formoterol, phenylethanolamine A are established. Data on optimization of hydrolysis, extraction and purification of extracts are given. Information on stage-by-stage losses of the determined compounds is given; data on the analysis of reference material are obtained. The validation of the developed method shows that the relative expanded uncertainty ranges from 7 to 24 %.

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About the authors

A. V. Sorokin

All-Russian State Center for Quality and Standardization of Drugs for Animals and Feeds (FGBU VGNKI)

Author for correspondence.
Email: alex_sorokin@list.ru
Russian Federation, Moscow

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Supplementary files

Supplementary Files
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2. Fig. 1. Examples of mass chromatograms of formoterol, phenylethanolamine A, and bambuterol in a liver sample at the limit of quantification.

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3. Fig. 2. Mass chromatograms of beta-agonists in a reference liver sample (a) without hydrolysis, (b) with hydrolysis using 30 µL of enzyme.

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4. Fig. 3. Comparison of peak areas of target compounds using different extractants. X-1 – 1% acetic acid in acetonitrile, X-3 – 1% ammonia solution in acetonitrile, X-4 – 0.1% acetic acid in acetonitrile.

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5. Fig. 4. Comparison of peak areas of target compounds during phase separation by salting out with different salts. S-2 – sodium sulfate and sodium chloride, S-3 – ammonium sulfate, S-4 – ammonium sulfate and sodium chloride.

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6. Fig. 5. Comparison of peak areas of target compounds using different purification methods. C-1 – 2 g of aluminum oxide and 1 g of C18, C-2 – 1 g of C18, C-3 – 2 g of aluminum oxide and 0.5 g of C18.

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7. Fig. 6. Comparison of average intensities of daughter ions of beta-agonists. CM-1 – multifunctional filter, CM-2 – separate extract purification.

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8. Fig. 7. Mass chromatograms of GM-clenbuterol and fenoterol at different spiking levels using multifunctional filter (CM-1) and separate extract purification (CM-2).

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9. Scheme 1. Structural formulas of formoterol (a), phenylethanolamine A (b), bambuterol (c).

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