Quantitative Determination of Four Azo Dyes in Rat Plasma with Solid-Phase Extraction and UFLC-MS-MS Analysis: Application to a Pharmacokinetic Study.
A rapid and sensitive ultra-fast liquid chromatography tandem mass spectrometry method, followed by simple protein precipitation and solid-phase extraction, has been developed and validated for the quantitative determination of four azo dyes (Para red, Solvent yellow 2, Solvent red 1 and Sudan red 7B) in rat plasma using D5-Sudan I as the internal standard. The optimal separation was accomplished on an Agilent Eclipse Plus C18 column (100 × 2.1 mm, 1.8 μm) with gradient elution using the mobile phase including acetonitrile and water (containing 0.1% formic acid). The flow rate was 0.45 mL/min. The detection was conducted by means of electrospray ionization mass spectrometry in positive ion mode with multiple reaction monitoring. The calibration curves showed good linearity, with correlation coefficients >0.998 for all of the analytes within the concentration range. The lower limits of quantification (LLOQs) of Para red, Solvent yellow 2, Solvent red 1 and Sudan red 7B in rat plasma were 1.0, 0.1, 0.1 and 0.1 μg/L, respectively. The intra- and interday relative standard deviations were ≤9.6 and ≤12.4%, respectively, and the accuracy was in the range of -5.8 to -9.5%. The average recoveries were between 81.49 and 118.65%, and the matrix effects were satisfactory in the biological matrices. The fully validated method has been successfully applied in measuring levels of the four azo dyes in rat plasma following oral administration of 20.0 mg/kg of analytes in rats, which was suitable for the pharmacokinetic studies of the azo dyes.
Department of Anesthesia, Ningbo First Hospital, Ningbo 315010, China.
Hao
H
Zhu
Department of Anesthesia, Ningbo First Hospital, Ningbo 315010, China.
Changshun
C
Huang
Department of Anesthesia, Ningbo First Hospital, Ningbo 315010, China.
Yijun
Y
Chen
Department of Anesthesia, Ningbo First Hospital, Ningbo 315010, China.
Zihui
Z
Lu
Department of Anesthesia, Ningbo First Hospital, Ningbo 315010, China.
Haidong
H
Zhou
Department of Anesthesia, Ningbo First Hospital, Ningbo 315010, China.
Chunru
C
Chen
Department of Anesthesia, Ningbo First Hospital, Ningbo 315010, China.
Jin
J
Wu
Zhejiang Provincial Key Laboratory of Health Risk Appraisal for Trace Toxic Chemicals, Ningbo Municipal Center for Disease Control and Prevention, Ningbo 315010, China.
Xiaohong
X
Chen
Zhejiang Provincial Key Laboratory of Health Risk Appraisal for Trace Toxic Chemicals, Ningbo Municipal Center for Disease Control and Prevention, Ningbo 315010, China.
Micong
M
Jin