Abstract
Tongmai formula (TMF) is a herbal preparation composed of three traditional Chinese medicinal materials: Puerariae Lobatae Radix (Gegen), Salviae Miltiorrhizae Radix et Rhizoma (Danshen) and Chuanxiong Rhizoma (Chuanxiong). It has been used to treat cardiovascular diseases for decades. To develop a reliable and convenient analytical method for a comprehensive determination of polyphenols in TMF and the ascertainment of their chemical correlations with its herbal components, a method combining high-performance liquid chromatography with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) was developed and validated for rapid determination of 30 polyphenols in TMF and its three herbal components. The chromatographic separation was carried out on a Chromolith Fastgradient RP-18 endcapped 50-2 column using an optimized gradient elution. Statistical analysis of obtained data demonstrated that the method had a desirable linearity, precision, and accuracy, as well as excellent sensitivity. The obtained results indicated that, among the 30 polyphenols in TMF, 22 originated from Gegen, 6 originated from Danshen, and 2 originated from Chuanxiong. The major polyphenols in TMF have been identified as puerarin, mirificin, salvianolic acid B, salvianic acid A, 3’-hydroxypuerarin, 3’-methoxypuerarin, and salvianolic acid A, with a combined contribution of 19.2% of the preparation. The development and validation of this method will greatly facilitate future pharmacological studies of TMF and its herbal components, as well as polyphenols in cardiovascular therapies.
Highlights
Polyphenols are secondary metabolites widely existing in plants but cannot be synthesized by animals and human beings [1]
We report the development of a method combining high-performance liquid chromatography with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) to analyze the polyphenols in Tongmai formula (TMF) and its herbal components
Methanol (MeOH), acetonitrile (MeCN), and formic acid (FA) used in high-performance liquid chromatography (HPLC)-ESI-MS/MS analysis were of HPLC grade and were purchased from Fisher Scientific (Springfield, NJ, USA)
Summary
Polyphenols are secondary metabolites widely existing in plants but cannot be synthesized by animals and human beings [1]. The most abundant phenols in Chuanxiong, has been declared to be responsible for the efficacy of Chuanxiong on cardiovascular diseases It shows antioxidative activity in vitro, inhibits platelet aggregation, prevents thrombus formation [14], exerts anti-hypertension effects [15,16], and reduces both triglyceride and total cholesterol plasma levels in vivo [14,16]. High-performance liquid chromatography (HPLC) fingerprint spectra have shown that polyphenols are the main constituents in TMF [17] and over 20 kinds of isoflavones have been isolated and identified from TMF [18,19], their exact contents are not clear yet Due to their essential pharmacological activities, reliable and convenient analytical methods are highly desired for the studies of the polyphenols. Studies of TMF, its herbal components, and polyphenols in in cardiovascular cardiovascular therapies
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