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Quantitative determination of ginsenoside Rh2 in rat biosamples by liquid chromatography electrospray ionization mass spectrometry

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Abstract

Ginsenoside Rh2 is a “hot” natural compound with great potential as a new anti-cancer drug based on abundant pharmacological experiments. However, no systemic pharmacokinetic study of Rh2 was reported because current analysis methods could not fully meet the requirements. Thus, we developed a simple LC/MS method with highly improved sensitivities for the determination of Rh2 in rat plasma, bile, urine, feces and most tissues. The tissues and feces were firstly homogenized mechanically using buffer and methanol as the media, respectively. Plasma, bile, urine and tissue homogenates were extracted with diethyl ether for sample preparation. Feces homogenates were directly deproteinized with acetonitrile. The subsequent analysis procedures were performed on a Shimadzu LCMS2010A system (electrospray ionization single quadrupole mass analyzer), with an ODS column (150 mm × 2.0-mm i.d., 5 μm) plus a C18 guard column for separation and ammonium chloride (500 μmol) as mobile phase additive. The proportions of mobile phase were changed timely according to gradient programs. Chlorinated adducts of molecular ions [M + Cl]− of Rh2 at m/z 657.35 and internal standard digitoxin at m/z 799.55 were monitored in selective ion monitoring mode of negative ions. The method was validated to be accurate, precise and rugged with good linearity in all matrices, according to the FDA guidelines. The lower limits of quantitation in rat plasma, urine and feces were 0.2, 0.2 and 20 ng/mL respectively. Stability studies were also performed, indicating that there were no stability-related problems in the analytical procedure of Rh2. The proposed method was successfully applied to the preclinical pharmacokinetic research of Rh2 in rats, including plasma kinetics, tissue distribution and excretion studies.

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Abbreviations

LC/ESI/MS:

liquid chromatography electrospray ionization mass spectrometry

HPLC:

high-performance liquid chromatography

LLOQ:

lower limit of quantitation

LC/MS/MS:

liquid chromatography tandem mass spectrometry

LOD:

limit of detection

IS:

internal standard

QC:

quality control

CDL:

curved desolvation line

DC:

direct current

RF:

radiation frequency

SIM:

selective ion monitoring

FDA:

Food and Drug Administration

CV:

coefficient of variation

SPE:

solid-phase extraction

LLE:

liquid–liquid extraction

t 0 :

dead time

ME:

matrix effect

MRT:

mean residence time

T 1/2 :

elimination half life

AUC:

area under the concentration–time curve

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Acknowledgements

This research was supported by the National High Technology Foundation of China (“863” Projects, No.2003AA2Z347A and No.2005AA2Z3C70), the fund of Jiangsu Key Lab of Drug Metabolism and Pharmacokinetics (No.BM2001201), National Nature Science Fund (No. 30572228), and the Natural Science Foundation of Jiangsu Province (BK 2005098). The great assistance from the group of post-graduates in the Key Lab of Drug Metabolism & Pharmacokinetics, China Pharmaceutical University is highly appreciated.

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Authors and Affiliations

  1. Key Laboratory of Drug Metabolism and Pharmacokinetics, China Pharmaceutical University, 1 Shennong Road, Nanjing, 210038, China

    Yi Gu, Guang-Ji Wang, Jian-Guo Sun, Yuan-Wei Jia, Hai-Tang Xie & Wei Wang

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  1. Yi Gu
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  2. Guang-Ji Wang
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  3. Jian-Guo Sun
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  4. Yuan-Wei Jia
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  5. Hai-Tang Xie
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  6. Wei Wang
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Correspondence to Guang-Ji Wang.

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Gu, Y., Wang, GJ., Sun, JG. et al. Quantitative determination of ginsenoside Rh2 in rat biosamples by liquid chromatography electrospray ionization mass spectrometry. Anal Bioanal Chem 386, 2043–2053 (2006). https://doi.org/10.1007/s00216-006-0857-8

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  • DOI: https://doi.org/10.1007/s00216-006-0857-8

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