Simultaneous determination and pharmacokinetic study of eight components in rat plasma by UHPLC-MS/MS after oral administration of Hypericum japonicum Thunb extract

doi:10.1016/j.jpba.2015.10.027

Journal of Pharmaceutical and Biomedical Analysis

Volume 118, 25 January 2016, Pages 228-234

https://doi.org/10.1016/j.jpba.2015.10.027 Get rights and content

Highlights

•
Simultaneous determination of eight active constituents in rat plasma.
•
Pharmacokinetic study of eight active constituents by UHPLC-MS/MS.
•
The first report on pharmacokinetic study of Hypericum japonicum Thunb by UHPLC-MS/MS.

Abstract

A rapid and sensitive assay based on ultra high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) was established and validated for the simultaneous determination of gallic acid, protocatechuic acid, vanillic acid, caffeic acid, epicatechin, isoquercitrin, vincetoxicoside B and quercetin in rat plasma using catechin and daidzein as the internal standards (IS). Plasma samples added internal standards were acidified with formic acid then pretreated by direct protein precipitation with acetonitrile. The separation of eight constituents was achieved on a C₁₈ column with gradient elution using methanol and 0.2% acetic acid aqueous solution as the mobile phase and detected by multiple reaction monitoring using electrospray ionization source in the positive–negative ionization mode. The method was validated for sufficient specificity, precision, accuracy, and sensitivity over the concentration range of 10–6000 ng mL⁻¹ for gallic acid, 1.5–3000 ng mL⁻¹ for protocatechuic acid, 10–15000 ng mL⁻¹ for vanillic acid, 2–3600 ng mL⁻¹ for caffeic acid, 1.5–3600 ng mL⁻¹ for epicatechin, 4–6000 ng mL⁻¹ for isoquercitrin, 2–9000 ng mL⁻¹ for vincetoxicoside B, and 20–18000 ng mL⁻¹ for quercetin. The overall intra‑run precision and the inter‑run precision were showed in the range of 1.0–14.2% and 2.8–12.9%, respectively, and the accuracy was no more than 12.8%. This analytical method was successfully applied to investigate the pharmacokinetics of eight ingredients in rats after oral administration of Hypericum japonicum Thunb extract.

Graphical abstract

Introduction

Hypericum japonicum Thunb, a member of genus Hypericum L. (Clusiaceae/Hypericaceae), is an annual or perennial herb widely distributed in Asia, North America and Oceania [1]. It has been widely used for the therapy of the infectious hepatitis, hemostasis, detumescence, dysentery and relieving internal heat or fever [1], [2]. Many pharmacological studies have demonstrated that its dried entire plant possesses antibacterial, antioxidant, antihypoxic and hepatoprotective activities [3], [4], [5]. Phytochemical investigations of H. japonicum Thunb have indicated many ingredients such as flavonoids, phenolic acids, xanthones, phloroglucinols and chromones [6]. Among them, phenolic acids and flavonoids are the main bioactive constituents which contribute to the pharmacological efficacy in H. japonicum Thunb. Based on a tremendous amount of previous pharmacological researches, phenolic acids (gallic acid, protocatechuic acid, vanillic acid and caffeic acid) and flavonoids (epicatechin, isoquercitrin, vincetoxicoside B and quercetin) strikingly manifest a variety of pharmacological properties, including antioxidant, antimicrobial, anti-inflammatory and antitumour functions [7], [8], [9], [10], [11]. In addition, it has displayed good effect in clinical application of hepatitis [12].

Most of previous studies have been concentrated on the phytochemical and other pharmacological investigation of H. japonicum Thunb. However, apart from quercitrin and isoquercitrin [13], no research reports on pharmacokinetics of other components of H. japonicum Thunb was available. It is well known that Traditional Chinese Medicine (TCM) has thousands of years of history in China that play a crucial role in the disease protection, control, and treatment. But there is a considerable lack in therapeutic mechanism understanding due to its manifold and complicated constituents. Pharmacokinetic data may help to illustrate and foresee efficacy and toxicity of the herb medicine and to optimize dose regiment and reduce adverse effects [14].

In this study, a rapid, sensitive and robust ultra high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) was established for the simultaneous determination of gallic acid, protocatechuic acid, vanillic acid, caffeic acid, epicatechin, isoquercitrin, vincetoxicoside B and quercetin of H. japonicum Thunb in rat plasma using catechin and daidzein as the internal standards. The method was fully validated and successfully applied to the pharmacokinetics after oral administration of H. japonicum Thunb extract to rats. The results may supply some valuable references for the apprehension of pharmacological action mechanism of H. japonicum Thunb.

Section snippets

Chemicals, reagents and animals

The standards of gallic acid, protocatechuic acid, caffeic acid and epicatechin were purchased from National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). Standards substances of isoquercitrin, vincetoxicoside B, quercetin and catechin (IS) were obtained from Chengdu Pufeide Biological Technology Co., Ltd. (Sichuan, China). The standards of vanillic acid and daidzein (IS) were purchased from Sichuan Weikeqi Biological Technology Co., Ltd. (Sichuan,

Selectivity and specificity

The selectivity and specificity were assessed using independent plasma samples derived from six different rats. The typical chromatograms for the drug‑free plasma, drug‑free with analytes at LLOQ and IS and an in vivo rat plasma sample after oral administration of H. japonicum Thunb are shown in Fig. 1. As shown in Fig. 1, all the peaks can be confirmed and there is no significant interference from the endogenous substances observed at the retention time of each component.

Linearity and sensitivity

As for plasma

Conclusions

In the present study, a sensitive and reliable UHPLC-MS/MS method was established for simultaneous determination of eight components following oral administration of H. japonicum Thunb extract in rat plasma. The analysis assay is simple and fast because of the straightforward sample pre‑treatment procedure and its relative short analysis running time. This is the first report of pharmacokinetic studies of gallic acid, protocatechuic acid, vanillic acid, caffeic acid, epicatechin, isoquercitrin,

Acknowledgements

This research was supported by the National Natural Science foundation of China (No. 20865001), the Guangxi Province Natural Science foundation (0832034).

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LC–MS/MS determination and pharmacokinetic study of five flavone components after solvent extraction/acid hydrolysis in rat plasma after oral administration of Verbena officinalis L. extract
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Simultaneous determination of catechin, epicatechin and epicatechin gallate in rat plasma by LC–ESI–MS/MS for pharmacokinetic studies after oral administration of Cynomotium songaricum extract
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Highlights

Abstract

Graphical abstract

Introduction

Section snippets

Chemicals, reagents and animals

Selectivity and specificity

Linearity and sensitivity

Conclusions

Acknowledgements

References (18)

Evaluation of pharmacological properties and phenolic profile of Hypericum japonicum Thunb from Western Ghats of India

J. Pharm. Res.

Hepatoprotective effect of Hypericum japonicum extract and its fractions

J. Ethnopharmacol.

Phytochemical analysis and antioxidant activities of Lantana camara and Lantana motevidensis extracts

Ind. Crops Prod.

Pharmacokinetic comparisons of different combinations of Shaoyao–Gancao–Decoction in rats: simultaneous determination of ten active constituents by HPLC-MS/MS

J. Chromatogr. B

LC–MS/MS determination and pharmacokinetic study of five flavone components after solvent extraction/acid hydrolysis in rat plasma after oral administration of Verbena officinalis L. extract

J. Ethnopharmacol.

Simultaneous determination of catechin, epicatechin and epicatechin gallate in rat plasma by LC–ESI–MS/MS for pharmacokinetic studies after oral administration of Cynomotium songaricum extract

J. Chromatogr. B

Dictionary of Chinese Traditional Medicine (Zhong Yao Da Ci Dian)

A New phloroglucinol diglycoside derivative from Hypericum japonicum Thunb

Molecules

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The influence of essential oils from Xiang-Fu-Si-Wu Decoction on its non-volatile components and its application for pharmacokinetics in normal rats

Quantitative determination of a synthetic amide derivative of gallic acid, SG-HQ2, using liquid chromatography tandem mass spectrometry, and its pharmacokinetics in rats

Simultaneous determination of ten Aconitum alkaloids in rat tissues by UHPLC–MS/MS and its application to a tissue distribution study on the compatibility of Heishunpian and Fritillariae thunbergii Bulbus

Comparative Computational Screening of Natural-based Partial Agonists for PPARγ Receptor

Short communicationSimultaneous determination and pharmacokinetic study of eight components in rat plasma by UHPLC-MS/MS after oral administration of Hypericum japonicum Thunb extract

Highlights

Abstract

Graphical abstract

Introduction

Section snippets

Chemicals, reagents and animals

Selectivity and specificity

Linearity and sensitivity

Conclusions

Acknowledgements

J. Pharm. Res.

J. Ethnopharmacol.

Ind. Crops Prod.

J. Chromatogr. B

J. Ethnopharmacol.

J. Chromatogr. B

Dictionary of Chinese Traditional Medicine (Zhong Yao Da Ci Dian)

A New phloroglucinol diglycoside derivative from Hypericum japonicum Thunb

Molecules

Short communication
Simultaneous determination and pharmacokinetic study of eight components in rat plasma by UHPLC-MS/MS after oral administration of Hypericum japonicum Thunb extract