Simultaneous determination of 1-chloro-2,4-dinitrobenzene, 2,4-dinitrophenyl- S-glutathione and its metabolites for human placental disposition studies by high-performance liquid chromatography

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 859; no. 1; pp. 94 - 102
• Main Authors: Vaidya, Soniya S., Gerk, Phillip M.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01.11.2007; Elsevier Science
• Subjects: 1-Chloro-2,4-dinitrobenzene (CDNB); 2,4-Dinitrophenyl- S-glutathione (DNP-SG); ABCC2; ABCG2; Analysis; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Buffers; Chromatography, High Pressure Liquid - methods; Dinitrochlorobenzene - analysis; Dinitrochlorobenzene - metabolism; Female; Fundamental and applied biological sciences. Psychology; General pharmacology; Glutathione - analogs & derivatives; Glutathione - analysis; GSTP1-1; Humans; Medical sciences; Pharmacology. Drug treatments; Placenta - metabolism; Placental metabolism and transport; Pregnancy; Reference Standards; Reproducibility of Results; ABC; ABCC1; ABCC2; DNP; DNP-Cys; CDNB; MRP; Placental metabolism and transport; GSTP1-1; PCA; 1-Chloro-2,4-dinitrobenzene (CDNB); DNP-CG; FDNB; DNP-NAc; DPBS; ABCG2; 2,4-Dinitrophenyl- S-glutathione (DNP-SG); Human; Chemical analysis; l-Chloro-2,4-dinitrobenzene (CDNB); Fetal membrane; Enzyme; Isozyme; Metabolite; Transferases; Biological transport; Tripeptide; HPLC chromatography; Metabolism; 2,4-Dinitrophenyl-S-glutathione (DNP-SG); Placenta; Glutathione transferase; Sulfur peptide; Simultaneous measurement; Pharmacokinetics; Glutathione
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2007.09.006

We developed and validated an HPLC method for determination of 1-chloro-2,4-dinitrobenzene (CDNB) and its glutathione conjugate 2,4-dinitrophenyl- S-glutathione (DNP-SG) to study the kinetics and mechanisms involved in DNP-SG formation and efflux, as a probe for human placental metabolism and transport. This method combines use of 3 μm solid phase, rapid mobile phase gradient with dual wavelength ultraviolet detection to permit determination of a lipophilic parent compound and its hydrophilic metabolites in a single short run. The selectivity, linearity, accuracy, precision, relative recovery and stability of the assay are sufficient for determining CDNB, DNP-SG and its metabolites from buffer and tissue samples to support placental drug metabolism and transport studies.