Establishment of UPLC-PDA Fingerprint of Baoyuantang Substance Benchmark and Identification of Its Chemical Constituents / 中国实验方剂学杂志

Objective:To establish the fingerprint of Baoyuantang substance benchmark， and to analyze and identify the common peaks. Method:A total of 15 batches of Baoyuantang substance benchmark were prepared， ultra performance liquid chromatography-diode array detector method （UPLC-PDA） was used to establish the fingerprint of the substance benchmark， and the methodology was developed. The chromatographic conditions were as follows：ACQUITY UPLC BEH Shield C₁₈ column （2.1 mm×100 mm， 1.7 μm）， mobile phase of 0.05% formic acid solution （A） and 0.05% formic acid acetonitrile solution （ B） for gradient elution （0-0.5 min， 5%-19%B； 0.5-6 min， 19%B； 6-10 min， 19%-27%B； 10-20 min， 27%-45%B； 20-20.1 min， 45%-95%B； 20.1-23 min， 95%B）， the flow rate of 0.4 mL·min^-1， the column temperature of 30 ℃， the detection wavelength at 203 nm and 260 nm， and the injection volume of 2 μL. Similarity evaluation system of traditional Chinese medicine fingerprint （2012 edition） was used to establish the fingerprint and generate the control fingerprint. The chemical constituents of Baoyuantang substance benchmark were identified by comparison of standard substances and UPLC-electrospray ionization tandem mass spectrometry （UPLC-ESI-MS/MS） with full information tandem mass spectrometry （MS^E） and scanning range of <italic>m</italic>/<italic>z</italic> 50-1 200. Result:The similarities of 15 batches of Baoyuantang substance benchmark were above 0.90 by comparing with the control fingerprint. There were 37 common peaks， 22 of which were identified through UPLC-ESI-MS/MS， including liquiritin， violanthin， ginsenoside Rg₁， ginsenoside Rb₁， ginsenoside Re and so on. These components were all from Astragali Radix， Ginseng Radix et Rhizoma， Zingiberis Rhizoma Recens and Glycyrrhizae Radix et Rhizoma. Conclusion:This method is accurate， stable and reliable， which will basically reflect the overall chemical composition characteristics of Baoyuantang， and it provides experimental basis for development of the granules of this famous classical formulas.

Historical Evolution and Modern Research on Shaoyao Gancaotang / 中国实验方剂学杂志

Shaoyao Gancaotang, first seen in Treatise on Febrile and Miscellaneous Diseases, is composed of Paeoniae Radix Alba and Glycyrrhizae Radix et Rhizoma Praeparata cum Melle in equal proportion.It has the functions of preserving Yin to nourish blood, harmonizing liver and spleen, relieving spasm and pain.This formula is applied in leg and foot spasm and abdominal pain caused by blood deficiency, body fluid consuming and the unmoistened muscles and veins.It has been highly praised and used by medical experts throughout the ages and has extended its application scope.Modern pharmacological studies have shown that Shaoyao Gancaotang has significant effects in antispasmodic, analgesic, antitussive and other areas, and is used to treat spastic diseases, painful diseases, inflammatory diseases and so on.This paper will systematically elaborate the historical evolution, compatibility analysis, pharmacological and pharmacodynamic studies, modern clinical application of Shaoyao Gancaotang, in order to provide theoretical basis and reference for the development of this famous classical formula.

Application of DESI-MSI in Quality Control of Banxia Xiexintang / 中国实验方剂学杂志

Objective::To prepare 15 batches of Banxia Xiexintang substance benchmark and lyophilized powder from different places, and the lyophilized powder was analyzed by ultra-high performance liquid chromatography with diode array detection (UHPLC-DAD) and desorption electrospray ionization-mass spectrometry imaging (DESI-MSI) in order to investigate the advantages of DESI-MSI in quality control of famous classical formulas. Method::Taking Banxia Xiexintang as the research model, fingerprints of the substance benchmark and lyophilized powder were established by UHPLC-DAD, and the content of index components and the yield of dry extract were also investigated. Meanwhile, as the research carrier, the lyophilized powder corresponding to Banxia Xiexintang was dissolved in methanol and dotted on qualitative filter paper with 5 μL quantitative capillary, and fixed it on the slide to make samples. The samples were analyzed on a DESI-MSI system in positive and negative ion mode with methanol-formic acid (1 000∶1, flow rate of 3 μL·min-1) as spray solvent, N2 as spray gas (pressure of 0.5 MPa). The scanning range was 100-1 200 Da, the spatial resolution was 300 μm, the spray voltage was 3 kV, the sampling cone voltage was ±40 V, incidence angle of sprayer was 60 degree, its collection angle was 10 degree, the ion source temperature was 120 ℃. Result::DESI-MSI could not only detect the index components of liquiritin, baicalin and wogonoside, as well as the common peaks of liquiritin apioside, berberine and glycyrrhizic acid, but also analyzed them semi-quantitatively, the analysis results were basically consistent with UHPLC-DAD. At the same time, DESI-MSI could detect 16 other components from Glycyrrhizae Radix et Rhizoma, Coptidis Rhizoma, Scutellariae Radix, Jujubae Fructus and Ginseng Radix et Rhizoma, such as licoricesaponin G2, palmatine, coptisine, rutin and ginsenoside Rg1, and present their relative content visually. The qualitative analysis ability of DESI-MSI was much better than UHPLC-DAD. Conclusion::DESI-MSI can be used as the quality control method for substance benchmark and lyophilized powder and dispensing granules of famous classical formulas with advantages of high sensitivity, strong analytical ability, no complex sample processing, qualitative and relative content analysis of complex samples without reference substance.