Determination of furosemide and its glucuronide metabolite in plasma, plasma ultrafiltrate and urine by HPLC-MS/MS with application to secretion and metabolite formation clearances in non-pregnant and pregnant women

Author(s)

Benzi, Jhohann Richard de Lima ; Rocha, Adriana ; Colombari, Julia Cristina ; Pego, Alef Machado Gomes ; Melli, Patricia Pereira dos Santos ; Duarte, Geraldo ; Lanchote, Vera Lucia

• Document type: Journal article
• Source: Journal of Pharmaceutical and Biomedical Analysis; v. 235, p. 9-pg., 2023-10-25.

Abstract

Furosemide (FUR) has been used in probe drugs cocktails for in vivo evaluation of the renal transporters OAT1 and OAT3 activities in studies of drug-drug interactions (generally using probenecid as an inhibitor) and drug-disease interactions. The objective of this study was to develop and validate methods for FUR and its glucuronide metabolite (FUR-GLU) analysis in plasma, plasma ultrafiltrate and urine for application in pharmacokinetics studies: a pilot drug-drug interaction study in pregnant women (n = 2), who received a single oral dose of FUR (40 mg) and in another occasion a single oral dose of probenecid (750 mg) before a single oral dose of FUR (40 mg), and in non-pregnant women participants (n = 12), who only received a single oral dose of FUR (40 mg). The samples preparation for FUR in 50 mu L of plasma and plasma lysate were carried by acidified liquid-liquid extraction, while 50 mu L of urine and 200 mu L of plasma ultrafiltrate were simply diluted with the mobile phase. The methods presented linearities in the range of 0.50 - 2500 ng/mL of plasma and plasma lysate, 0.125 - 250 ng/mL of plasma ultrafiltrate, and 50 - 20,000 ng/mL of urine. FUR-GLU methods presented linearities in the range of 0.125 - 250 ng/mL of plasma ultrafiltrate and 50 - 20,000 ng/mL of urine. Precision and accuracy evaluations showed coefficients of variation and relative errors < 15%. In the pregnant women participants, the mean values of FUR CLrenal, CLsecretion, CLformation. FUR-GLU and CLnon- renal were all reduced when probenecid was administered with FUR (8.24 vs 2.89 L/h, 8.15 vs 2.80 L/h, 3.86 vs 1.75 L/h, 48.26 vs 22.10 L/h, respectively). Non-pregnant women presented similar values of FUR CL(rena)l, CLsecretion, CLformation. FUR-GLU to the pregnant women who received FUR only. Finally, FUR fraction unbound (fu) resulted in values of approximately 1% in pregnant women and to 0.22% in non-pregnant women. These developed and validated methods for FUR and FUR-GLU quantification in multiple matrices can allow the further investigation of UGT1A9/1A1 and the fu when FUR is administered as an OAT 1 and 3 in vivo probe. (AU)

• FAPESP's process: 21/10292-5 - Quantitative in vitro to in vivo prediction of the effects of pro-inflammatory cytokines and pregnancy on transporter-mediated drug disposition via PBPK modeling and simulation
• Grantee: Jhohann Richard de Lima Benzi
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• FAPESP's process: 18/05616-3 - Clinical pharmacokinetics in infectious diseases
• Grantee: Vera Lúcia Lanchote
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 21/00351-4 - Impact of pyelonephritis on the activity of organic anion transporters (OAT) 1 and 3 in pregnant women: a quantitative approach to systems pharmacology
• Grantee: Julia Cristina Colombari
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• FAPESP's process: 19/03429-4 - Pyelonephritis impacts on Organic Anion Transporters (OAT) 1 and 3 activity in pregnant women: a quantitative system pharmacology approach
• Grantee: Jhohann Richard de Lima Benzi
• Support Opportunities: Scholarships in Brazil - Doctorate