A sensitive LC-MS/MS method for the quantitative determination of biflorin in rat plasma and its application to pharmacokinetic studies.
A rapid, sensitive and selective liquid chromatography-tandem mass spectrometric method (LC-MS/MS) was developed for the quantification of biflorin in rat plasma. Using naringin as an internal standard, plasma samples were subjected to a direct protein precipitation process using methanol. Chromatographic separation was achieved on a Gemini C18 column with an isocratic mobile phase consisting of 0.1% formic acid and methanol (50:50, v/v) at a flow rate of 0.5mL/min. Biflorin was analyzed in the multiple reaction monitoring mode with negative electrospray ionization. The precursor/product ion pairs were m/z 353.0/205.0 and m/z 579.0/271.0 for biflorin and the IS, respectively. The calibration curve was linear over the concentration range of 5-2000ng/mL. The intra- and inter-day precision was less than 7.3% and the accuracy ranged from 96.5 to 103.3%. No significant variation was observed in the stability tests. This method was successfully applied to a pharmacokinetic study of biflorin after the intravenous and oral administration of biflorin to rats. The half-life and oral bioavailability of biflorin were determined as 3.4h and 43%, respectively. This is the first report on the quantitative determination of biflorin in rat plasma as well as the pharmacokinetic characterization of biflorin, which should provide a meaningful foundation for further preclinical and clinical applications of biflorin.
BK21 Plus Project Team, College of Pharmacy, Dongguk University-Seoul, Goyang, Republic of Korea.
Seung Jun
SJ
Yang
College of Pharmacy, Kyung Hee University, Seoul, Republic of Korea.
Jong Hoon
JH
Ryu
Department of Life and Nanopharmaceutical Science, Kyung Hee University, Seoul, Republic of Korea.
Dae Sik
DS
Jang
BK21 Plus Project Team, College of Pharmacy, Dongguk University-Seoul, Goyang, Republic of Korea.
Liang
L
Yang
BK21 Plus Project Team, College of Pharmacy, Dongguk University-Seoul, Goyang, Republic of Korea. Electronic address: hkhan@dongguk.edu.
Hyo-Kyung
HK
Han
D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D007275:Injections, Intravenous; D057230:Limit of Detection; D008297:Male; D009285:Naphthoquinones / Q000008:administration & dosage / Q000097:blood* / Q000302:isolation & purification; D010936:Plant Extracts / Q000008:administration & dosage / Q000302:isolation & purification / Q000493:pharmacokinetics*; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
D009285:Naphthoquinones; D010936:Plant Extracts; C522781:biflorin
Biflorin; LC–MS/MS; Pharmacokinetics; Plasma; Rat