Generic placeholder image

Current Pharmaceutical Analysis


ISSN (Print): 1573-4129
ISSN (Online): 1875-676X

Research Article

A Novel UPLC-MS/MS Method for the Determination of Salvianolic Acid A in Rat Urine, Feces, and Bile and its Application to Excretion Study

Author(s): Jialin Sun, Wen Xu, Junke Song, Xiao Li, Qie Guo, Wei Sun, Bing Han, Guanhua Du* and Fanbo Jing*

Volume 15, Issue 3, 2019

Page: [266 - 272] Pages: 7

DOI: 10.2174/1573412914666171211160018


Background: Salvianolic acid A (SAA) is a polyphenolic acid extracted from Salvia miltiorrhiza Bunge. It showed protective effect against diabetic complications after oral administration with a low bioavailability of 1.42%. Attempts have been made to develop it into a new medication. Intracorporal process of SAA is indistinct and no report regarding the excretion is available. Our preliminary experiment revealed that previous reported methods were unsuitable for the excretion study due to the serious matrix effect.

Methods: To better clarify its pharmacokinetics and avoid the interference of complex endogenous substances, a sensitive UPLC-MS/MS method with a better resolution was developed for the excretion study of SAA for the first time. The analytes were separated by reversed-phase chromatography with acetonitrile-water (containing 0.1% formic acid) gradient elution. The mass spectrometer was operated in the negative ESI mode and multiple reaction monitoring mode.

Results: This method was linear over the concentration range of 2.5-100, 5-100 and 5-100 ng/mL in urine, feces and bile, respectively. The accuracy, precision, stability, recovery and matrix effect were satisfactory in all matrices examined. The validated method was successfully applied to an excretion study in rats. After oral administration of 20 mg/kg, the average accumulated excretion amount of SAA in urine, feces and bile were 99.80, 32046.30 and 161.03 ng, respectively.

Conclusion: A quick but low elimination was observed. The date is useful for the clinical trial design of SAA.

Keywords: Salvianolic acid A, UPLC-MS/MS, rat, excretion, sensitivity, resolution.

Graphical Abstract
Li, Y.; Shi, S.; Gao, J.; Han, S.; Wu, X.; Jia, Y.; Su, L.; Shi, J.; Hu, D. Cryptotanshinone downregulates the profibrotic activities of hypertrophic scar fibroblasts and accelerates wound healing: A potential therapy for the reduction of skin scarring. Biomed. Pharmacother., 2016, 80, 80-86.
Shang, Q.; Xu, H.; Huang, L. Tanshinone IIA: A promising natural cardioprotective agent. Evid. Based Complement. Alternat. Med., 2012, 2012, 716459.
Zhou, L.; Zuo, Z.; Chow, M.S. Danshen: an overview of its chemistry, pharmacology, pharmacokinetics, and clinical use. J. Clin. Pharmacol., 2005, 45(12), 1345-1359.
Xie, Z.; Loi Truong, T.; Zhang, P.; Xu, F.; Xu, X.; Li, P. Dan-Qi prescription ameliorates insulin resistance through overall corrective regulation of glucose and fat metabolism. J. Ethnopharmacol., 2015, 172, 70-79.
Gao, L.N.; Yan, K.; Cui, Y.L.; Fan, G.W.; Wang, Y.F. Protective effect of Salvia miltiorrhiza and Carthamus tinctorius extract against lipopolysaccharide-induced liver injury. World J. Gastroenterol., 2015, 21(30), 9079-9092.
Jian, W.; Yu, S.; Tang, M.; Duan, H.; Huang, J. A combination of the main constituents of Fufang Xueshuantong Capsules shows protective effects against streptozotocin-induced retinal lesions in rats. J. Ethnopharmacol., 2016, 182, 50-56.
Guo, X.J.; Jiang, Z.; Zhao, P.F.; Fang, L.L.; Hu, X.Y.; Wang, P.; Guo, X. Pharmacokinetic study of four components in rat plasma after oral administration of Guanmaitong Granule by UPLC-MS/MS. Curr. Pharm. Anal., 2018, 14(3), 223-232.
Si, Y.; Li, N.; Tong, L.; Lin, B.; Wang, W.; Xing, Y.; Liu, X.; Yue, H.; Xu, J.; Ju, A. Bioactive minor components of the total salvianolic acids injection prepared from Salvia miltiorrhiza Bge. Bioorg. Med. Chem. Lett., 2016, 26(1), 82-86.
Ren-an, Q.; Juan, L.; Chuyuan, L.; Wenjuan, F.; Chunyan, H.; Xuemei, Y.; Lin, H.; Hong, N. Study of the protective mechanisms of Compound Danshen Tablet (Fufang Danshen Pian) against myocardial ischemia/reperfusion injury via the Akt-eNOS signaling pathway in rats. J. Ethnopharmacol., 2014, 156, 190-198.
Hu, P.; Liang, Q.L.; Luo, G.A.; Zhao, Z.Z.; Jiang, Z.H. Multi-component HPLC fingerprinting of Radix Salviae Miltiorrhizae and its LC-MS-MS identification. Chem. Pharm. Bull. (Tokyo), 2005, 53(6), 677-683.
Liu, M.; Li, Y.G.; Zhang, F.; Yang, L.; Chou, G.X.; Wang, Z.T.; Hu, Z.B. Chromatographic fingerprinting analysis of Danshen root (Salvia miltiorrhiza Radix et Rhizoma) and its preparations using high performance liquid chromatography with diode array detection and electrospray mass spectrometry (HPLC-DAD-ESI/MS). J. Sep. Sci., 2007, 30(14), 2256-2267.
Huang, J.; Qin, Y.; Liu, B.; Li, G.Y.; Ouyang, L.; Wang, J.H. In silico analysis and experimental validation of molecular mechanisms of salvianolic acid A-inhibited LPS-stimulated inflammation, in RAW264.7 macrophages. Cell Prolif., 2013, 46(5), 595-605.
Fan, H.Y.; Fu, F.H.; Yang, M.Y.; Xu, H.; Zhang, A.H.; Liu, K. Antiplatelet and antithrombotic activities of salvianolic acid A. Thromb. Res., 2010, 126(1), e17-e22.
Xu, X.; Hu, Y.; Zhai, X.; Lin, M.; Chen, Z.; Tian, X.; Zhang, F.; Gao, D.; Ma, X.; Lv, L.; Yao, J. Salvianolic acid A preconditioning confers protection against concanavalin A-induced liver injury through SIRT1-mediated repression of p66shc in mice. Toxicol. Appl. Pharmacol., 2013, 273(1), 68-76.
Wu, Z.M.; Wen, T.; Tan, Y.F.; Liu, Y.; Ren, F.; Wu, H. Effects of salvianolic acid a on oxidative stress and liver injury induced by carbon tetrachloride in rats. Basic Clin. Pharmacol. Toxicol., 2007, 100(2), 115-120.
Wang, S.B.; Tian, S.; Yang, F.; Yang, H.G.; Yang, X.Y.; Du, G.H. Cardioprotective effect of salvianolic acid A on isoproterenol-induced myocardial infarction in rats. Eur. J. Pharmacol., 2009, 615(1-3), 125-132.
Xu, T.; Wu, X.; Chen, Q.; Zhu, S.; Liu, Y.; Pan, D.; Chen, X.; Li, D. The anti-apoptotic and cardioprotective effects of salvianolic acid a on rat cardiomyocytes following ischemia/reperfusion by DUSP-mediated regulation of the ERK1/2/JNK pathway. PLoS One, 2014, 9(7), e102292.
Qiang, G.; Yang, X.; Shi, L.; Zhang, H.; Chen, B.; Zhao, Y.; Zu, M.; Zhou, D.; Guo, J.; Yang, H.; Zhang, L.; Du, G. Antidiabetic effect of salvianolic acid a on diabetic animal models via AMPK activation and mitochondrial regulation. Cell. Physiol. Biochem., 2015, 36(1), 395-408.
Wu, P.; Yan, Y.; Ma, L.L.; Hou, B.Y.; He, Y.Y.; Zhang, L.; Niu, Z.R.; Song, J.K.; Pang, X.C.; Yang, X.Y.; Du, G.H. Effects of the nrf2 protein modulator salvianolic acid A alone or combined with metformin on diabetes-associated macrovascular and renal injury. J. Biol. Chem., 2016, 291(42), 22288-22301.
Yu, X.; Zhang, L.; Yang, X.; Huang, H.; Huang, Z.; Shi, L.; Zhang, H.; Du, G. Salvianolic acid A protects the peripheral nerve function in diabetic rats through regulation of the AMPK-PGC1alpha-Sirt3 axis. Molecules, 2012, 17(9), 11216-11228.
Yang, X.Y.; Qiang, G.F.; Zhang, L.; Zhu, X.M.; Wang, S.B.; Sun, L.; Yang, H.G.; Du, G.H. Salvianolic acid A protects against vascular endothelial dysfunction in high-fat diet fed and streptozotocin-induced diabetic rats. J. Asian Nat. Prod. Res., 2011, 13(10), 884-894.
Yang, X.Y.; Sun, L.; Xu, P.; Gong, L.L.; Qiang, G.F.; Zhang, L.; Du, G.H. Effects of salvianolic scid A on plantar microcirculation and peripheral nerve function in diabetic rats. Eur. J. Pharmacol., 2011, 665(1-3), 40-46.
Sun, J.; Zhang, L.; Song, J.; Tian, S.; Huang, C.; Feng, Z.; Lv, Y.; Du, G. Pharmacokinetic study of salvianolic acid A in beagle dog after oral administration by a liquid chromatography-mass spectrometry method: a study on bioavailability and dose proportionality. J. Ethnopharmacol., 2013, 148(2), 617-623.
Hou, Y.Y.; Peng, J.M.; Chao, R.B. Pharmacokinetic study of salvianolic acid A in rat after intravenous administration of Danshen injection. Biomed. Chromatogr., 2007, 21(6), 598-601.
Zhang, X.R.; Pan, W.S. Quantitative determination of salvianolic acid A in rat plasma by HPLC. Chinese Traditional and Herbal Drugs, 2002, 33(5), 410-412.
Zhao, D.; Han, D.E.; Li, N.; Lu, Y.; Li, T.T.; Yang, S.Y.; He, J.K.; Chen, X.J. Simultaneous determination of six phenolic constituents of Danshen injection in rat plasma by LC-ESI-MS and its application to a pharmacokinetic study. Eur. J. Mass Spectrom. (Chichester), 2011, 17(4), 395-403.
Chang, B.B.; Zhang, L.; Cao, W.W.; Cao, Y.; Yang, W.L.; Wang, Y.; Chen, Y.C.; Liu, X.Q. Pharmacokinetic interactions induced by content variation of major water-soluble components of Danshen preparation in rats. Acta Pharmacol. Sin., 2010, 31(5), 638-646.
Li, X.; Du, F.; Jia, W.; Olaleye, O.E.; Xu, F.; Wang, F.; Li, L. Simultaneous determination of eight Danshen polyphenols in rat plasma and its application to a comparative pharmacokinetic study of DanHong injection and Danshen injection. J. Sep. Sci., 2017, 40(7), 1470-1481.
Pei, L.; Bao, Y.; Wang, H.; Yang, F.; Xu, B.; Wang, S.; Yang, X.; Du, G. A sensitive method for determination of salvianolic acid A in rat plasma using liquid chromatography/tandem mass spectrometry. Biomed. Chromatogr., 2008, 22(7), 786-794.
Wang, H.; Li, X.; Zhang, W.; Liu, Y.; Wang, S.; Liu, X.; He, H. Mechanism-based pharmacokinetic-pharmacodynamic modeling of salvianolic acid A effects on plasma xanthine oxidase activity and uric acid levels in acute myocardial infarction rats. Xenobiotica, 2017, 47(3), 208-216.
Wang, S.P.; Liu, L.; Wang, L.L.; Jiang, P.; Xiang, L.; Zhang, W.D.; Liu, R.H. Simultaneous determination of six hydrophilic components in rat plasma after oral administration of Jitai tablet by liquid chromatography-electrospray ionization-tandem mass spectrometry: application to a pharmacokinetic study. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci., 2013, 912, 75-84.
Du, , G.H.; Zhang, W.K; Zhang, L.. ; Zhao, Y.; Wang, S.B.; Yang, H.G.; Tian, S A method of preparation of salvianolic acid A CN102731308A (in Chinese)., 2012.
Han, D.E.; Zheng, Y.; Chen, X.; He, J.; Zhao, D.; Yang, S.; Zhang, C.; Yang, Z. Identification and characterization of human UDP-glucuronosyltransferases responsible for the in vitro glucuronidation of salvianolic acid A. Drug Metab. Pharmacokinet., 2012, 27(6), 579-585.
Xu, J.; Zeng, S.; Chen, X.; Qu, H. Isolation and identification of degradation products of salvianolic acid A by NMR and LC-MS. Fitoterapia, 2011, 82(2), 260-266.

© 2024 Bentham Science Publishers | Privacy Policy