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DEVELOPMENT AND VALIDATION OF HPLC-MS/MS METHODS FOR THE DETERMINATION OF THYMODEPRESSIN IN THE SERUM OF BLOOD RABBITS

Authors: Stepanova E.S.; Makarenkova L.M.; Abramovich R.A.; Chistyakov V.V.;

DEVELOPMENT AND VALIDATION OF HPLC-MS/MS METHODS FOR THE DETERMINATION OF THYMODEPRESSIN IN THE SERUM OF BLOOD RABBITS

Abstract

The HPLC-MS/MS method for the determination of gamma-D-glutamyl-D-tryptophan (γ-HT) in serum was developed and validated. Target transition in MS/MS mode for γ-HT was m/z 334.0→188.1. The extraction of γ-HT from the serum was carried out by acetonitrile precipitation of the proteins with further extraction using methylene chloride. The calibration curve in the concentration range 10-5000 ng/ml was linear, the correlation coefficient was >0.999. The limit of detection was 5.0 ng/ml, the limit of quantification was 10 ng/ml. The developed method was used for quantitative determination of γ-HT in the serum of blood rabbits which received a new dosage form of Thymodepressin - a powder for inhalation.

Разработана и валидирована методика ВЭЖХ-МС/МС определения гамма-О-глутамил-D-триптофана (γ-ГТ) в сыворотке крови. Целевой переход в МС/МС режиме m/z 334,0 → 188,1. Извлечение γ-ГТ из сыворотки проводили осаждением белков ацетонитрилом с экстракцией хлористым метиленом. Калибровочная кривая в диапазоне концентраций 10-5000 нг/ мл линейна, коэффициент корреляции >0,999. Предел обнаружения - 5,0 нг/мл, предел количественного определения - 10 нг/мл. Разработанную методику использовали для количественного определения γ-ГТ в сыворотке крови кроликов, получавших новую лекарственную форму препарата «Тимодепрессин» - порошка для ингаляции.

Keywords

validation, пептидный иммунодепрессант, HPLC-MS/MS, гамма-D-глутамил-D-триптофан, "Тимодепрессин", ВЭЖХ-МС/МС, валидация, peptide immunosuppressant, gamma-D-glutamyl-D-tryptophan, thymodepressin

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
0
Average
Average
Average
Green