A rapid and sensitive UFLC-MS/MS method for the simultaneous determination of gentiopicroside and swertiamarin in rat plasma and its application in pharmacokinetics.
OBJECTIVE
Radix Gentianae is a traditional Chinese medicine derived from medicinal plants of the family Gentianaceae. Its pharmacological effects have been primarily attributed to the presence of a number of secoiridoid glycosides, in particular gentiopicroside and swertiamarin. In this study, a rapid and sensitive method based on ultrafast liquid chromatography-tandem mass spectrometry has been developed for the simultaneous determination of gentiopicroside and swertiamarin in rat plasma using paeoniflorin as internal standard (IS).


METHODS
After liquid-liquid extraction with ethyl acetate-isopropanol (95 : 5, v/v), separation was achieved on a Shim-pack XR-ODS C18 column (75 mm × 3.0 mm, 2.2 μm) with a mobile phase consisting of methanol : 0.1% formic acid (30 : 70, v/v) at a flow rate of 0.4 ml/min. Detection on an API 3200 QTRAP mass spectrometer equipped with an electrospray ionization source operated in the negative ionization mode was performed by multiple reaction monitoring of the precursor-to-product ion transitions of gentiopicroside, swertiamarin and IS at m/z 401.0 → 179.0, 419.0 → 179.1 and 525.1 → 121.0 respectively. The calibration curves were linear over the concentration range of 20-10 000 and 2-1000 ng/ml for gentiopicroside and swertiamarin with corresponding lower limits of quantification of 20 and 2 ng/ml. The limits of detection were 4 and 0.5 ng/ml for gentiopicroside and swertiamarin, respectively. The intraday and interday precisions were below 11.9% for gentiopicroside and below 9.5% for swertiamarin in terms of relative standard deviation, and the accuracy was within ±8.3% for gentiopicroside and within ±10.2% for swertiamarin in terms of relative error. Extraction recovery, matrix effect and stability were satisfactory in rat plasma. The method was fully validated and applied to a pharmacokinetic study involving oral administration of a Radix Gentianae extract to groups of male and female rats.


RESULTS
Results showed that in female rats, both compounds were absorbed to a greater extent and eliminated more slowly than in male rats, although the rate of absorption was similar in the two groups.


CONCLUSIONS
There were remarkable differences in pharmacokinetic properties of gentiopicroside and swertiamarin between male and female rats. The results will provide helpful information for the development of suitable dosage forms and clinical references on rational administration.
Research Center for Drug Metabolism, College of Life Science, Jilin University, China; School of Pharmacy, Jilin Medical College, China.
Bo
B
Feng
Heyun
H
Zhu
Jiao
J
Guan
Longshan
L
Zhao
Jingkai
J
Gu
Lei
L
Yin
J Paul
JP
Fawcett
Wenshu
W
Liu
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004365:Drugs, Chinese Herbal / Q000737:chemistry / Q000493:pharmacokinetics*; D005260:Female; D030012:Gentiana / Q000737:chemistry*; D007408:Intestinal Absorption; D057888:Iridoid Glucosides / Q000097:blood / Q000493:pharmacokinetics*; D008297:Male; D018517:Plant Roots; D011753:Pyrones / Q000097:blood / Q000493:pharmacokinetics*; D017207:Rats, Sprague-Dawley; D012737:Sex Factors; D053719:Tandem Mass Spectrometry / Q000379:methods
D004365:Drugs, Chinese Herbal; D057888:Iridoid Glucosides; D011753:Pyrones; C012997:gentiopicroside; C013270:swertiamarin
Radix Gentianae; UFLC-MS/MS; gentiopicroside; pharmacokinetics; swertiamarin