Drug bioavailability evaluation
Solutions of seven drugs (Antipyrine, Propranolol hydrochloride,
Hydrochlorothiazide, (±)-Sulpiride, and Ranitidine hydrochloride) were
purchased from Shanghai Aladdin Biochemical Technology. Atenolol and
Furosemide were purchased from Dalian Meilune Biochemical Technology.
The drugs were prepared at a concentration of 2 μmol/ml in HBSS from
Meilunbio. Prior to drug absorption evaluation, cells were washed twice
with HBSS for 30 minutes. Then, 150 μL of the drug solution was added to
the middle well, while 600 μL of HBSS was added to the lower well. After
a 3-hour incubation, 500 μL of medium from the lower well was collected
and centrifuged at 8000 rpm for 10 minutes. The supernatant was
quantified by liquid chromatography (LC20A, Shimadzu, Japan) with a
chromatographic column size of 250 mm*4.6 mm, 5 μm, C18. The injection
volume was 10 μL. Antipyrine was detected at a wavelength of 254 nm,
Propranolol hydrochloride at 223 nm, Verapamil at 278 nm,
Hydrochlorothiazide at 270 nm, Ranitidine hydrochloride at 230 nm,
Atenolol at 240 nm, (±)-Sulpiride at 230 nm, and Furosemide at 279 nm.