Bioanalytical Method Validation of an RP-HPLC Method for Determination of Rifampicin in Liver Perfusion Studies

Background: The number of validated quantification methods for rifampicin, a prototypical Oatp inhibitor, in biological rat samples was limited. Objective: This study was conducted to validate a modified reversed-phase liquid chromatographic method for the determination of rifampicin in rat liver tissue according to the current ICH M10 Bioanalytical Method Validation Draft Guideline (2019) for application to samples of in situ rat liver perfusion studies. Method: Liver tissue samples were obtained from recirculatory in situ rat liver perfusion studies. The analysis was performed on a C18 column with a mobile phase composed of 0.05 M phosphate buffer (pH 4.58): acetonitrile (55:45, v/v). The assay was validated for selectivity, calibration curve and range, matrix effect, carry-over, accuracy and precision, reinjection reproducibility, and stability. Results: The method was considered selective and stable, without having carry-over and matrix effects. The calibration curve was linear (R2: 0.9983) within the calibration range (0.5-60 ppm). Accuracy and precision values fulfilled the required limits. Liver concentrations of rifampicin in liver tissue, obtained after 60 min perfusion with 10 µM and 50 µM of rifampicin were 45.1 ± 11.2 and 313.4 ± 84.4 µM, respectively. Conclusion: The bioanalytical method validation was completed and the method was successfully applied for the determination of rifampicin in rat liver tissue.

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Effects of Perfusate on Cation Transport by Slices from the Isolated Perfused Rat Liver

Canadian Journal of Physiology and Pharmacology ◽

10.1139/y72-130 ◽

1972 ◽

Vol 50 (9) ◽

pp. 916-919

Author(s):

A. C. Nestruck ◽

R. W. Furneaux

Keyword(s):

Red Blood Cells ◽

Rat Liver ◽

Blood Cells ◽

Liver Perfusion ◽

Isolated Perfused Rat Liver ◽

Buffer Solution ◽

Bicarbonate Buffer ◽

Rat Liver Perfusion ◽

Metabolic Substrates ◽

Perfusion Studies

Isolated livers from fed rats were perfused for 1 h with a basic Krebs–Ringer bicarbonate buffer solution containing albumin and glucose and added (1) α-ketoglutarate, (2) pyruvate, fumarate, and glutamate, or (3) washed red blood cells. Perfusate flow rate, [Formula: see text], [Formula: see text], and pH changes across the liver, and glucose efflux in the perfusate, were measured during perfusion. Rewarmed slices of liver taken at the beginning of perfusion were found to be able to reverse a cation shift imposed by cold incubation. Slices of liver taken after 1 h of perfusion were not able to effect this cation shift unless red blood cells were included in the perfusate. It is proposed that noncellular perfusates containing metabolic substrates are not ideal for isolated rat liver perfusion studies as evidenced in the altered membrane transport capacity of slices after perfusion.

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Bioanalytical method validation for determination of arsenic speciation in dog plasma using HPLC-ICP/MS

Analytical Science and Technology ◽

10.5806/ast.2016.29.5.234 ◽

2016 ◽

Vol 29 (5) ◽

pp. 234-241

Author(s):

Jong-Hwan Kim ◽

Young Sang Kwon ◽

Min-Chul Shin ◽

Su Jong Kim ◽

Jong-Su Seo

Keyword(s):

Method Validation ◽

Arsenic Speciation ◽

Bioanalytical Method Validation ◽

Bioanalytical Method ◽

Dog Plasma ◽

Icp Ms

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Bioanalytical Method Validation for the Determination of Warfarin in Spiked-Saliva Using Fluorometric HPLC for TDM Application

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.1162.173 ◽

2021 ◽

Vol 1162 ◽

pp. 173-179

Author(s):

Ari Wibowo ◽

Shabrina Nurbaiti ◽

Vitarani Dwi Ananda Ningrum

Keyword(s):

Method Validation ◽

Mobile Phase ◽

Geriatric Patients ◽

Therapeutic Drug ◽

Excitation Wavelength ◽

Bioanalytical Method Validation ◽

Bioanalytical Method ◽

Non Invasive ◽

Accuracy And Precision

Saliva becomes an alternative biological matrix for therapeutic drug monitoring (TDM) application since there is a strong correlation between warfarin plasma concentration and saliva; further, the sampling is non-invasive and more comply with pediatric and geriatric patients. This study aims to validate the parameters of the warfarin bioanalytical method in spiked-saliva according to the criteria from the Food and Drug Administration (FDA) in the Guidance for Industry Bioanalytical Method Validation. The method used is Fluorometric HPLC with an excitation wavelength of 310 nm and an emission wavelength of 390 nm. The mobile phase involved is phosphate buffer-methanol, and the stationary phase is C18. The LoD and LoQ obtained are 0.71 ng/mL and 2.16 ng/mL, respectively. The coefficient of variation and %diff in the selectivity, accuracy, and precision parameters have met the criteria of the bioanalytical method of less than 20%. Meanwhile, the average %recovery is 101.30%. To conclude, the developed warfarin bioanalytical method has fulfilled the established criteria. It can, therefore, be used to determine warfarin concentration in saliva as an alternative method for TDM services in the clinical domain.

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