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Frontiers of Chemical Science and Engineering

ISSN 2095-0179

ISSN 2095-0187(Online)

CN 11-5981/TQ

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Front Chem Sci Eng    2012, Vol. 6 Issue (4) : 436-442    https://doi.org/10.1007/s11705-012-1222-4
RESEARCH ARTICLE
LC/MS/MS and radioisotope method combined for recognizing the affinity between catalpol and OCT2 transporter
Zhiyu ZHANG1,2, Changxiao LIU1,2, Duanyun SI2, Rong LU2, Xiulin YI2()
1. School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China; 2. State Key Laboratory of Drug Delivery Technology and Pharmacokinetics, Tianjin Institute of Pharmaceutical Research, Tianjin 300193, China
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Abstract

More and more herbal medicines are found to be the substrates of drug transporters. In this paper, chromatography/tandem mass spectrometry (LC/MS/MS) combined with radioisotope method was used for the quantification of catalpol, a traditional Chinese medicine, to study the affinity relationship between herbal medicines and transporters. Catalpol uptake experiment was carried out by using several transporters (OAT1, OCT2, OAT3, OATP1B1 and OATP2B1). And samples were precipitated with methanol and quantified with LC/MS/MS. The results show that catalpol has a good affinity with OCT2-transfected S2 cells. After studying drug-drug interaction between catalpol and 14C-tetraethylammonium (TEA), we found that catalpol is able to facilitate TEA transport mediated by OCT2, suggesting that catalpol could probably be a new promoter of OCT2.
Keywords catalpol      LC/MS/MS      OCT2     
Corresponding Author(s): YI Xiulin,Email:yixl@tjipr.com   
Issue Date: 05 December 2012
 Cite this article:   
Zhiyu ZHANG,Changxiao LIU,Duanyun SI, et al. LC/MS/MS and radioisotope method combined for recognizing the affinity between catalpol and OCT2 transporter[J]. Front Chem Sci Eng, 2012, 6(4): 436-442.
 URL:  
https://academic.hep.com.cn/fcse/EN/10.1007/s11705-012-1222-4
https://academic.hep.com.cn/fcse/EN/Y2012/V6/I4/436
Fig.1  Representative chromatogram of IS and catalpol
Fig.1  Representative chromatogram of IS and catalpol
Fig.1  Representative chromatogram of IS and catalpol
Fig.1  Representative chromatogram of IS and catalpol
Fig.1  Representative chromatogram of IS and catalpol
Fig.1  Representative chromatogram of IS and catalpol
Fig.2  (a) 5 μmol/L C-PAH, OAT1, uptake time: 2 min, inhibitor: 1 mmol/L non-radiolabeled PAH; (b) 60 μmol/L C-TEA, OCT2, uptake time: 2 min, inhibitor: 1 mmol/L non-radiolabeled TEA; (c) 50 nmol/L 3H-estrone sulfate, OAT3, uptake time: 2 min, inhibitor: 300 μmol/L non-radiolabeled estrone sulfate; (d) 50 nmol/L 3H-estrone sulfate, OATP1B1, uptake time: 1 min, inhibitor: 30 μmol/L non-radiolabeled estrone sulfate; (e) 50 nmol/L 3H-estrone sulfate, OATP2B1, uptake time: 1 min, inhibitor: 300 μmol/L non-radiolabeled estrone sulfate
Cell function confirmation
Fig.2  (a) 5 μmol/L C-PAH, OAT1, uptake time: 2 min, inhibitor: 1 mmol/L non-radiolabeled PAH; (b) 60 μmol/L C-TEA, OCT2, uptake time: 2 min, inhibitor: 1 mmol/L non-radiolabeled TEA; (c) 50 nmol/L 3H-estrone sulfate, OAT3, uptake time: 2 min, inhibitor: 300 μmol/L non-radiolabeled estrone sulfate; (d) 50 nmol/L 3H-estrone sulfate, OATP1B1, uptake time: 1 min, inhibitor: 30 μmol/L non-radiolabeled estrone sulfate; (e) 50 nmol/L 3H-estrone sulfate, OATP2B1, uptake time: 1 min, inhibitor: 300 μmol/L non-radiolabeled estrone sulfate
Cell function confirmation
Fig.2  (a) 5 μmol/L C-PAH, OAT1, uptake time: 2 min, inhibitor: 1 mmol/L non-radiolabeled PAH; (b) 60 μmol/L C-TEA, OCT2, uptake time: 2 min, inhibitor: 1 mmol/L non-radiolabeled TEA; (c) 50 nmol/L 3H-estrone sulfate, OAT3, uptake time: 2 min, inhibitor: 300 μmol/L non-radiolabeled estrone sulfate; (d) 50 nmol/L 3H-estrone sulfate, OATP1B1, uptake time: 1 min, inhibitor: 30 μmol/L non-radiolabeled estrone sulfate; (e) 50 nmol/L 3H-estrone sulfate, OATP2B1, uptake time: 1 min, inhibitor: 300 μmol/L non-radiolabeled estrone sulfate
Cell function confirmation
Fig.2  (a) 5 μmol/L C-PAH, OAT1, uptake time: 2 min, inhibitor: 1 mmol/L non-radiolabeled PAH; (b) 60 μmol/L C-TEA, OCT2, uptake time: 2 min, inhibitor: 1 mmol/L non-radiolabeled TEA; (c) 50 nmol/L 3H-estrone sulfate, OAT3, uptake time: 2 min, inhibitor: 300 μmol/L non-radiolabeled estrone sulfate; (d) 50 nmol/L 3H-estrone sulfate, OATP1B1, uptake time: 1 min, inhibitor: 30 μmol/L non-radiolabeled estrone sulfate; (e) 50 nmol/L 3H-estrone sulfate, OATP2B1, uptake time: 1 min, inhibitor: 300 μmol/L non-radiolabeled estrone sulfate
Cell function confirmation
Fig.2  (a) 5 μmol/L C-PAH, OAT1, uptake time: 2 min, inhibitor: 1 mmol/L non-radiolabeled PAH; (b) 60 μmol/L C-TEA, OCT2, uptake time: 2 min, inhibitor: 1 mmol/L non-radiolabeled TEA; (c) 50 nmol/L 3H-estrone sulfate, OAT3, uptake time: 2 min, inhibitor: 300 μmol/L non-radiolabeled estrone sulfate; (d) 50 nmol/L 3H-estrone sulfate, OATP1B1, uptake time: 1 min, inhibitor: 30 μmol/L non-radiolabeled estrone sulfate; (e) 50 nmol/L 3H-estrone sulfate, OATP2B1, uptake time: 1 min, inhibitor: 300 μmol/L non-radiolabeled estrone sulfate
Cell function confirmation
Fig.2  (a) 5 μmol/L C-PAH, OAT1, uptake time: 2 min, inhibitor: 1 mmol/L non-radiolabeled PAH; (b) 60 μmol/L C-TEA, OCT2, uptake time: 2 min, inhibitor: 1 mmol/L non-radiolabeled TEA; (c) 50 nmol/L 3H-estrone sulfate, OAT3, uptake time: 2 min, inhibitor: 300 μmol/L non-radiolabeled estrone sulfate; (d) 50 nmol/L 3H-estrone sulfate, OATP1B1, uptake time: 1 min, inhibitor: 30 μmol/L non-radiolabeled estrone sulfate; (e) 50 nmol/L 3H-estrone sulfate, OATP2B1, uptake time: 1 min, inhibitor: 300 μmol/L non-radiolabeled estrone sulfate
Cell function confirmation
Fig.3  (a) OAT1; (b) OCT2; (c) OAT3; (d) OATP1B1; (e) OATP2B1
Catalpol uptake investigation
Fig.3  (a) OAT1; (b) OCT2; (c) OAT3; (d) OATP1B1; (e) OATP2B1
Catalpol uptake investigation
Fig.3  (a) OAT1; (b) OCT2; (c) OAT3; (d) OATP1B1; (e) OATP2B1
Catalpol uptake investigation
Fig.3  (a) OAT1; (b) OCT2; (c) OAT3; (d) OATP1B1; (e) OATP2B1
Catalpol uptake investigation
Fig.3  (a) OAT1; (b) OCT2; (c) OAT3; (d) OATP1B1; (e) OATP2B1
Catalpol uptake investigation
Fig.3  (a) OAT1; (b) OCT2; (c) OAT3; (d) OATP1B1; (e) OATP2B1
Catalpol uptake investigation
Fig.4  Drug-drug interaction Sixty μmol/L C-TEA mediated by OCT2 was used as positive control (middle); 30 μmol/L TEA, 100 μmol/L TEA, 300 μmol/L TEA, 30 μmol/L catalpol, 100 μmol/L catalpol and 300 μmol/L catalpol were added additionally for studying the variation of C-TEA uptake. Unpaired -test was done with GraphPad Prism 3.0 software. μM=μmol?L, * <0.05; ** <0.01; *** <0.001
Fig.4  Drug-drug interaction Sixty μmol/L C-TEA mediated by OCT2 was used as positive control (middle); 30 μmol/L TEA, 100 μmol/L TEA, 300 μmol/L TEA, 30 μmol/L catalpol, 100 μmol/L catalpol and 300 μmol/L catalpol were added additionally for studying the variation of C-TEA uptake. Unpaired -test was done with GraphPad Prism 3.0 software. μM=μmol?L, * <0.05; ** <0.01; *** <0.001
Fig.4  Drug-drug interaction Sixty μmol/L C-TEA mediated by OCT2 was used as positive control (middle); 30 μmol/L TEA, 100 μmol/L TEA, 300 μmol/L TEA, 30 μmol/L catalpol, 100 μmol/L catalpol and 300 μmol/L catalpol were added additionally for studying the variation of C-TEA uptake. Unpaired -test was done with GraphPad Prism 3.0 software. μM=μmol?L, * <0.05; ** <0.01; *** <0.001
Fig.4  Drug-drug interaction Sixty μmol/L C-TEA mediated by OCT2 was used as positive control (middle); 30 μmol/L TEA, 100 μmol/L TEA, 300 μmol/L TEA, 30 μmol/L catalpol, 100 μmol/L catalpol and 300 μmol/L catalpol were added additionally for studying the variation of C-TEA uptake. Unpaired -test was done with GraphPad Prism 3.0 software. μM=μmol?L, * <0.05; ** <0.01; *** <0.001
Fig.4  Drug-drug interaction Sixty μmol/L C-TEA mediated by OCT2 was used as positive control (middle); 30 μmol/L TEA, 100 μmol/L TEA, 300 μmol/L TEA, 30 μmol/L catalpol, 100 μmol/L catalpol and 300 μmol/L catalpol were added additionally for studying the variation of C-TEA uptake. Unpaired -test was done with GraphPad Prism 3.0 software. μM=μmol?L, * <0.05; ** <0.01; *** <0.001
Fig.4  Drug-drug interaction Sixty μmol/L C-TEA mediated by OCT2 was used as positive control (middle); 30 μmol/L TEA, 100 μmol/L TEA, 300 μmol/L TEA, 30 μmol/L catalpol, 100 μmol/L catalpol and 300 μmol/L catalpol were added additionally for studying the variation of C-TEA uptake. Unpaired -test was done with GraphPad Prism 3.0 software. μM=μmol?L, * <0.05; ** <0.01; *** <0.001
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