[Simultaneous determination of baicalin and chlorogenic acid in human plasma by UPLC-MS/MS].
To develop and validate an ultra performance liquid chromatography tandem mass spectrometric (UPLC-MS/MS) method for the simultaneous quantification of baicalin and chlorogenic acid in human plasma after iv infusion of Yinhuang injection, the analytes were isolated from plasma by protein precipitation with methanol. Then they were chromatographied on an Acquity UPLC BEH C18 column (50 mm x 2.1 mm ID, 1.7 microm) at 40 degrees C. The mobile phase A consisted of water and 0.1% formic acid. The mobile phase B consisted of methanol and 0.1% formic acid. The analytes were eluted from the column with a linear gradient from 5% B to 80% B in 5 min, then hold for 0.5 min before returning to initial condition. The flow rate was 0.35 mL x min(-1). A tandem mass spectrometer equipped with electrospray ionization source was used as detector. Multiple reaction monitoring (MRM) using the precursor to product ion pairs of m/z 447-->271 (for baicalin), m/z 353-->191 (for chlorogenic acid) and m/z 287-->287 (for internal standard) were used to quantification. The linear concentration ranges of the calibration curves for baicalin and chlorogenic acid ranged from 9.6 to 1540 ng x mL(-1) and from 7.5 to 1200 ng x mL(-1), respectively. The intra- and inter-day relative standard deviation (RSD) across three validations run over the entire concentration range was less than 10.2% for both baicalin and chlorogenic acid. After iv infusion of Yinhuang injection to the volunteers, the concentration-time curves of baicalin and chlorogenic acid fitted the two-compartment and three-compartment model. T(1/2)alpha were (4.47 +/- 0.89) and (7.65 +/- 4.42) min, T(1/2)beta were (46.22 +/- 10.03) and (34.40 +/- 19.16) min, respectively. The method was proved to be highly sensitive, selective, and suitable for pharmacokinetic investigations of both baicalin and chlorogenic acid.
Ju
Wen-zheng
WZ
Department of Clinical Pharmacology, Jiangsu Provincial Hospital of Traditional Chinese Medicine, Nanjing University of Traditional Chinese Medicine, Nanjing 210029, China. njwz1008@jlonline.com
Liu
Fang
F
Wu
Ting
T
Xia
Xiao-yan
XY
Tan
Heng-shan
HS
Wang
Xing
X
Xiong
Ning-ning
NN
D019540:Area Under Curve; D002726:Chlorogenic Acid / Q000097:blood* / Q000493:pharmacokinetics; D002853:Chromatography, Liquid / Q000379:methods; D004338:Drug Combinations; D004365:Drugs, Chinese Herbal / Q000008:administration & dosage / Q000302:isolation & purification / Q000493:pharmacokinetics*; D005260:Female; D005419:Flavonoids / Q000097:blood* / Q000493:pharmacokinetics; D006801:Humans; D007275:Injections, Intravenous; D031239:Lonicera / Q000737:chemistry; D008297:Male; D010946:Plants, Medicinal / Q000737:chemistry; D031364:Scutellaria baicalensis / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods; D055815:Young Adult
D004338:Drug Combinations; D004365:Drugs, Chinese Herbal; D005419:Flavonoids; D002726:Chlorogenic Acid; C038044:baicalin