Determination of berberine in human plasma by liquid chromatography–electrospray ionization–mass spectrometry

doi:10.1016/j.jpba.2007.03.022

Journal of Pharmaceutical and Biomedical Analysis

Volume 44, Issue 4, 15 August 2007, Pages 931-937

https://doi.org/10.1016/j.jpba.2007.03.022 Get rights and content

Abstract

A liquid chromatography–electrospray ionization–mass spectrometry (LC–ESI–MS) method for the determination of berberine in human plasma using chlorobenzylidine as the internal standard (IS) has been developed and validated. The plasma samples were prepared by LLE and the analytes were chromatographically separated on a Hanbon Lichrospher 5-C18 HPLC column under gradient elution with a mobile phase consisted of acetonitrile and 10 mm ammonium acetate buffer containing 0.1% formic acid. Berberine was determined with electrospray ionisation–mass spectrometry (ESI–MS). LC–ESI–MS was performed in the selected-ion monitoring (SIM) mode using target ions at M⁺ m/z 336.1 for berberine and M⁺ m/z 464.1 for the IS. Calibration curve was linear over the range of 0.020–3.0 ng/ml. The lower limit of quantification (LLOQ) was 0.020 ng/ml. The intra- and inter-run variability values were less than 6.7 and 7.7%, respectively. The method has been successfully applied to determine the plasma concentration of berberine in healthy Chinese volunteers.

Introduction

Berberine (Fig. 1A), a well-known alkaloid, was found in medicinal herbs such as Coptis chinensis and Hydrastis Canadensis. Currently, the predominant clinical uses of berberine preparations include the treatment of bacterial diarrhea, intestinal parasite infections and ocular trachoma infections [1]. Other pharmacological effects of berberine, such as antiarrhythmic [2], anti-inflammatory [3], anticancer [4], immunosuppressive [5], vasorelaxant and antiproliferative [6] have also been reported. Several methods have been reported for the qualitative and quantitative analysis of berberine in herbs and berberine preparations, which include thin-layer chromatography (TLC) [7], capillary electrophoresis coupled with laser-induced native fluorescence [8], capillary electrophoresis with electrospray mass spectrometry (CE-MS) [9] and ion-pair supercritical fluid chromatography (IP-SFC) on-line coupled with ion-pair supercritical fluid extraction (IP-SFE) [10]. The methods reported for the determination of berberine in animal biological specimen include TLC [11], HPLC [12], [13], [14] and LC–ESI–MS [15] methods, in which the most sensitive method is LC–ESI–MS with an LLOQ of 0.31 ng/ml. Recently, Zuo et al. [16] reported an more sensitive LC/MS–MS method for the determination of berberine in rat plasma with an LLOQ of 0.05 ng/ml. Though there are almost one hundred of papers published about the determination of berberine, very few methods are reported for the determination of berberine in human plasma. Miyazaki et al. [17] reported a GC–MS method with an LLOQ of 1 ng/ml for the determination of berberine in human urine. Yu et al. [18] developed an HPLC method for the determination of berberine in human urine and plasma, in which the limit of detection (LOD) was 0.1 ng/ml. In this paper, they reported a valuable data obtained in their pilot study that the steady-state concentration of berberine in human plasma was about 0.3 ng/ml after an oral multi-dose of 300 mg berberine per day for 7 days. The pilot pharmacokinetic study results in our laboratory showed that the C_max of berberine in human plasma was about 0.4 ng/ml and the concentration levels of berberine on the terminal elimination phase were below 0.1 ng/ml after the single oral dose of 400 mg berberine. So, to evaluate the pharmacokinetics of berberine in human, a more sensitive method for the determination of berberine in human plasma is required. The objective of this study is to develop and validate an LC–ESI–MS method for the measurement of berberine in human plasma in support of the clinical investigations. In this study, we established a more sensitive method with the linear range of 0.020–3.0 ng/ml. The method was fully validated for its accuracy, precision, recovery, stability studies and matrix effect, and successfully applied to study the pharmacokinetics of berberine in healthy volunteers.

Section snippets

Materials and reagents

Berberine hydrochloride was purchased from Chinese National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). Chlorobenzylidine hydrochloride was obtained from Medicinal Chemistry Research Center of China Pharmaceutical University. The test berberine hydrochloride tablet containing 0.1 g berberine per tablet was provided by Yunnan Gulin Pharmaceutical Co., Ltd. (Yunnan, China). And the reference berberine hydrochloride tablet containing 0.1 g berberine per

Method development and optimization

The goal of this study was to develop and validate a sensitive method for the extraction and quantification of berberine in human plasma, which is suitable for determination of pharmacokinetics of berberine in clinical study. To achieve the goal, during method development, different options were evaluated to optimize MS parameters, chromatography condition and sample extraction method.

Conclusions

In the study presented here, we report the development and validation of an LC–ESI–MS method for measurement of berberine in human plasma. Validation results demonstrated that this method is specific, reproducible and reliable. The basic underlying advantage of this method is that the LLOQ for berberine is as low as 0.020 ng/ml. No significant interferences and matrix effect caused by endogenous compounds were observed. The method is suitable for pharmacokinetic study of berberine in human

References (23)

N. Mitani et al.
Cancer Lett.
(2001)
E. Marinova et al.
Immunopharmacology
(2000)
W. Ko et al.
Eur. J. Pharmacol.
(2000)
Y. Chen et al.
J. Chromatogr. A
(2000)
C. Chen et al.
J. Chromatogr. B Biomed. Appl.
(1995)
P. Tsai et al.
J. Chromatogr. A
(2002)
T. Lu et al.
J. Pharm. Biomed. Anal.
(2006)
H. Miyazaki et al.
J. Chromatogr.
(1978)
L. Ding et al.
J. Pharm. Biomed. Anal.
(2006)
L. Ding et al.
J. Pharm. Biomed. Anal.
(2006)

L. Ding et al.

J. Pharm. Biomed. Anal.

(2007)

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Determination of berberine in human plasma by liquid chromatography–electrospray ionization–mass spectrometry

Abstract

Introduction

Section snippets

Materials and reagents

Method development and optimization

Conclusions

Cancer Lett.

Immunopharmacology

Eur. J. Pharmacol.

J. Chromatogr. A

J. Chromatogr. B Biomed. Appl.

J. Chromatogr. A

J. Pharm. Biomed. Anal.

J. Chromatogr.

J. Pharm. Biomed. Anal.

J. Pharm. Biomed. Anal.

J. Pharm. Biomed. Anal.