Development of a solvent-free analytical method for paracetamol quantitative determination in Blood Brain Barrier in vitro model.
A Reversed Phase-High Performance Liquid Chromatography/Diode Array Detection method was developed and validated for paracetamol quantification in cell culture fluid from an in vitro Blood Brain Barrier model. The chromatographic method and sample preparation were developed using only aqueous solvents. The column was a XTerra RP18 150 × 4.6mm, 3.5 μm with a guard column XTerra RP18 20 × 4.6 mm, 3.5 μm at 35 °C and the mobile phase was composed by 100% formate buffer 20 mM at pH 4 and flow rate was set at 1 mL/min. The detection was at 242 nm. The sample was injected at 10 μL. Validation was performed using the accuracy profile approach. The analytical procedure was validated with the acceptance limits at ± 10% over a range of concentration from 1 to 58 mg L(-1). The procedure was then used in routine to determine paracetamol concentration in a brain blood barrier in vitro model. Application of the Unither paracetamol formulation in Blood Brain Barrier model allowed the determination and comparison of the transcellular passage of paracetamol at 37 °C and 4 °C, that excludes paracellular or non specific leakage.
EA 4575 Analytical and Pharmaceutical Developments Applied to Neglected Diseases and Counterfeits, Univ. Bordeaux, Bordeaux, France.
Marie-Hélène
MH
Langlois
INSERM U1049« Neuroinflammation, Imagerie et Thérapie de la Sclérose en plaques », Univ. Bordeaux, Bordeaux, France.
Antonios
A
Vekris
Unither développement Bordeaux, avenue Toussaint Catros, ZA Tech Espace, 33185 Haillan, France.
Christine
C
Bousses
INSERM U1049« Neuroinflammation, Imagerie et Thérapie de la Sclérose en plaques », Univ. Bordeaux, Bordeaux, France.
Elodie
E
Mordelet
EA 4575 Analytical and Pharmaceutical Developments Applied to Neglected Diseases and Counterfeits, Univ. Bordeaux, Bordeaux, France.
Nathalie
N
Buhannic
EA 4575 Analytical and Pharmaceutical Developments Applied to Neglected Diseases and Counterfeits, Univ. Bordeaux, Bordeaux, France.
Céline
C
Séguard
P-O Couraud, INSERM Centre de Recherche Institut Cochin 22 rue Méchain, 75014 Paris, France.
Pierre-Olivier
PO
Couraud
Babette B. Weksler, Weill Cornell Medical College, New York City, New York, USA.
Babette B
BB
Weksler
INSERM U1049« Neuroinflammation, Imagerie et Thérapie de la Sclérose en plaques », Univ. Bordeaux, Bordeaux, France.
Klaus G
KG
Petry
EA 4575 Analytical and Pharmaceutical Developments Applied to Neglected Diseases and Counterfeits, Univ. Bordeaux, Bordeaux, France. Electronic address: Karen.gaudin@u-bordeaux.fr.
Karen
K
Gaudin
D000082:Acetaminophen / Q000032:analysis* / Q000493:pharmacokinetics*; D001812:Blood-Brain Barrier / Q000166:cytology / Q000378:metabolism*; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D004355:Drug Stability; D006801:Humans; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000082:Acetaminophen
Accuracy profile; Acetaminophen; Blood Brain Barrier in vitro model; Paracetamol; RP-HPLC; Solvent free