Simultaneous detection of four flavonoids and two alkaloids in rat plasma by LC-MS/MS and its application to a comparative study of the pharmacokinetics between Abri Herba and Abri mollis Herba extract after oral administration.

Abri Herba and Abri mollis Herba both were important members of the Leguminosae family in southwestern China. Abri mollis Herba was often used as Abri Herba due to their proximity, but there are few studies on pharmacokinetics to compare their main identical active compositions. A sensitive and selective high-performance liquid chromatography with tandem mass spectrometry method in the positive/negative electrospray ionization switching mode was developed and validated for the simultaneous analysis of four flavonoids and two alkaloids in rat plasma. The chromatographic separation was carried out on a C18 column with a gradient mobile phase consisting of methanol and 0.5% acetic acid. The detection of the target compounds was conducted in multiple-reaction monitoring mode with a hybrid triple quadrupole linear ion trap mass spectrometer equipped with positive/negative ion-switching electrospray ion source. The differences in pharmacokinetics were discovered, which indicated that the substitution between them is some degree of irrationality. The validated method was successfully applied to pharmacokinetic study of the six components in male rat plasma after oral administration of Abri Herba and Abri mollis Herba extract and the results in the study would provide a useful guide for the clinical application of Abri Herba with those in Abri mollis Herba.

Simultaneous Detection of Flavonoids, Phenolic Acids and Alkaloids in Abri Herba and Abri Mollis Herba using Liquid Chromatography Tandem Mass Spectrometry.

INTRODUCTION: Abri Herba has remarkable properties, such as cleanup heat detoxification, dampness and activating blood circulation to dissipate blood stasis; as a result, it has been applied to treat acute or chronic hepatitis and mastitis. Abri mollis Herba is often used as Abri Herba. Hierarchical cluster analysis (HCA) was applied to compare the similarities and differences of the chemical compositions in the two types of medicinal materials. OBJECTIVE: To establish a high-performance liquid chromatography and tandem mass spectrometry (HPLC-MS/MS) method for the simultaneous analysis of 15 flavonoids, two phenolic acids and three alkaloids in Abri Herba and Abri mollis Herba. METHODOLOGY: The chromatographic separation was performed on a C18 column with a mobile phase of methanol (A), acetonitrile (B) and 0.5‰ acetic acid in water (C) using gradient elution. The detection of the target compounds was performed in multiple-reaction monitoring (MRM) mode using a hybrid quadrupole linear ion trap mass spectrometer equipped with positive/negative ion-switching electrospray ionisation (ESI) source. RESULTS: The developed method is reliable, sensitive and specific. In addition, the method has been successfully applied to differentiate 15 batches of Abri Herba and 27 batches of Abri mollis Herba stems. Furthermore, a comparison of the contents among stems, roots and leaves from the same strain in seven batches of Abri mollis Herba and four batches of Abri Herba has also been performed. CONCLUSION: HPLC-MS/MS method is sensitive and selective and can be suitable for the reliable quality control of Abri mollis Herba and Abri Herba.

Simultaneous quantification of 25 active constituents in the total flavonoids extract from Herba Desmodii Styracifolii by high-performance liquid chromatography with electrospray ionization tandem mass spectrometry.

A sensitive and selective high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry method has been developed and validated for the simultaneous determination of 25 active constituents, including 21 flavonoids and four phenolic acids in the total flavonoids extract from Herba Desmodii Styracifolii for the first time. Among the 25 compounds, seven compounds including caffeic acid, acacetin, genistein, genistin, diosmetin, diosmin and hesperidin were identified and quantified for the first time in Herba Desmodii Styracifolii. Chromatographic separation was accomplished on a ZORBAX SB-C18 (250 mm×4.6 mm, 5.0 µm) column using gradient elution of methanol and 0.1‰ acetic acid v/v at a flow rate of 1.0 mL/min. The identification and quantification of the analytes were achieved using negative electrospray ionization mass spectrometry in multiple-reaction monitoring mode. The method was fully validated in terms of limits of detection and quantification, linearity, precision and accuracy. The results indicated that the developed method is simple, rapid, specific and reliable. Furthermore, the developed method was successfully applied to quantify the 25 active components in six batches of total flavonoids extract from Herba Desmodii Styracifolii.

Determination and Quantitative Comparison of Nucleosides in two Cordyceps by HPLC-ESI-MS-MS.

A hybrid quadruple linear ion trap liquid chromatography-tandem mass spectrometry (LC-MS-MS) analytical method has been developed for simultaneous determination of 13 nucleosides from Cordyceps cicadae and Cordyceps sinensis for assessing whether C. cicadae can become a substitute for C. sinensis, based on that C. cicadae has in common with C. sinensis for treating chronic diseases. Among the 13 compounds, three compounds including cytidine, hypoxanthine and 2'-deoxyguanosine were firstly identified and quantified in C. cicadae. Ideal separation was achieved in one single LC-MS-MS run of 12 min by optimized chromatographic conditions. The identification and quantification analysis of target compounds were performed in electrospray ionization tandem mass spectrometry. The limit of detection and quantity for 13 compounds were <42.0 and 84.2 ng/mL, respectively. The determination results of 12 batches of C. sinensis and 20 batches of C. cicadae were then analyzed and classified by multivariate statistical analysis. C. cicadae contained a relatively higher level of three nucleosides (cytidine, uracil and 2'-deoxyuridine) than those in C. sinensis. The results of this study provided the theoretical basis for the substitution of C. sinensis with C. cicadae. The optimized method could be used for the quality control and investigate bioactive compound variation of Cordyceps.