Screening for 12 mixed illegally cough-relieving chemical composition in anti-cough and antiasthmatic traditional Chinese medicines by HPLC-DAD-MS/MS / 药学学报

This study was designed to establish a high performance liquid chromatography (diode array detector, DAD)-tandem mass chromatography (HPLC-DAD-MS/MS) method for the simultaneously screening 12 kinds of cough-relieving chemical drugs illegally added in anti-cough and antiasthmatic traditional Chinese medicines. This method involved liquid chromatography-tandem mass spectrometry. The separation was conducted by Kromasil100-5C18 (250 mm × 4.6 mm, 5 μm) and the mobile phase was consisted of 0.3% ammonium formate (pH 2.97) and methanol. After separated by HPLC, the suspected components were analyzed by MS/MS and DAD and ultra scan was used to identify these illegally added drugs. A fast and sensitive HPLC-DAD-MS/MS method for the simultaneously screening for illegal chemical compositions was established. The LOD of these substances were below 50 ng. The method was sufficiently selective and sensitive to detect illegal chemical compositions in anti-cough and antiasthmatic traditional Chinese medicines.

HPLC-DAD-MS/MS profiling of phenolics from Securigera securidaca flowers and its anti-hyperglycemic and anti-hyperlipidemic activities

Abstract Securigera securidaca (L.) Degen & Döefl., Fabaceae, has been widely used in the Iranian, Indian and Egyptian folk medicine as antidiabetic and anti-hyperlipidemic remedy. Phenolic profiling of the ethanolic extract (90%) of the flowers of S. securidaca was performed via HPLC-DAD-MS/MS analysis in the positive and negative ion modes. The total polyphenols and flavonoids in the flowers were determined colorimetrically, and the quantification of their components was carried out using HPLC-UV. Total phenolics and flavonoids estimated as gallic acid and rutin equivalents were 82.39 ± 2.79 mg/g and 48.82 ± 1.95 mg/g of the dried powdered flowers, respectively. HPLC-DAD-MS/MS analysis of the extract allowed the identification of 39 flavonoids and eight phenolic acids. Quantitative analysis of some flavonoids and phenolics (mg/100 g powdered flowers) revealed the presence of isoquercetrin (3340 ± 2.1), hesperidin (32.09 ± 2.28), naringin (197.3 ± 30.16), luteolin (10.247 ± 0.594), chlorogenic acid (84.22 ± 2.08), catechin (3.94 ± 0.57) and protocatechuic acid (34.4 ± 0.15), in the extract. Moreover, the acute toxicity, hypoglycemic and hypolipidemic effects of the extract were investigated using alloxan induced diabetes in rats in a dose of 100, 200, and 400 mg/kg bwt. The ethanolic extract was safe up to a dose of 2000 mg/kg. All tested doses of the flower extract showed marked decrease in blood glucose level by 31.78%, 66.41% and 63.8% at 100, 200 and 400 mg/kg bwt, respectively, at p < 0.05. Regarding the anti-hyperlipidemic effect, a dose of 400 mg/kg of the flower extract showed the highest reduction in serum triacylglycerides and total cholesterol levels (68.46% and 51.50%, respectively at p < 0.05). The current study proved the folk use of the flowers of S. securidaca as anti-diabetic and anti-hyperlipidemic agent which could be attributed to its high phenolic content.

Analysis on Chemical Constituents of Chinese Materia Medica Formulation Sini Decoction by HPLC-DAD-MS/MS / 中草药·英文版

Objective: To develop a high-performance liquid chromatography (HPLC) tandem mass spectrometry method for screening and identifying the multiple constituents in the complex Chinese materia medica (CMM) formulation Sini Decoction. Methods: The chemical constituents in Sini prescription as an entirety were isolated and analyzed by HPLC coupled with electrospray ionization ion-trap mass spectrometry (ESI-IT-MS) in order to clarify the chemical components of Sini Decoction and explore the chemical methods of studying CMM prescriptions. Results: Twenty-five compounds were isolated from the water extract of this decoction. Through chemical and spectral analyses, the structures of 20 compounds were elucidated, including 15 flavonoids, one triterpene, two coumarins, and two other types of compounds. By using HPLC-DAD-MSn technique, the MSn cleavage behaviors of the flavonoids have been summarized as well as the origin of the isolated compounds was assigned. Conclusions: The phytochemical compositon of Sini prescription is tentatively characterized by HPLC-DAD-ESI-IT-MSn. The results of the experiments demonstrate that the combination of HPLC and ESI-MSn is a sensitive and effective tool for rapidly investigating the complex CMM formulations.

Quantitative and qualitative analysis of common peaks in chemical fingerprint of Yuanhu Zhitong tablet by HPLC-DAD-MS/MS / 药物分析学报

A quality control (QC) strategy for quantitative and qualitative analysis of “common peaks” in chemical fingerprint was proposed to analyze Yuanhu Zhitong tablet (YZT), using high performance liquid chromatography with diode array detector and tandem mass spectrometry (HPLC-DAD-MS/MS). The chromatographic separation was achieved on an Agilent Eclipse plus C18 column with a gradient elution using a mixture of 0.4‰ ammonium acetate aqueous (pH 6.0 adjusted with glacial acetic acid) and acetonitrile. In chemical fingerprint, 40 peaks were assigned as the “common peaks”. For quantification of “common peaks”, the detection wavelength was set at 254 nm, 270 nm, 280 nm and 345 nm, respectively. The method was validated and good results were obtained to simultaneously determine 10 analytes (protopine, jatrorrhizine, coptisine, palmatine, berberine, xanthotoxin, bergapten, tetrahydropalmatine, imperatorin and isoimperatorin). For qualification of “common peaks”, 33 compounds including 10 quantitative analytes were identified or tentatively characterized using LC-MS/MS. These results demonstrated that the present approach may be a powerful and useful tool to tackle the complex quality issue of YZT.

Bitter acids from hydroethanolic extracts of Humulus lupulus L., Cannabaceae, used as anxiolytic

Humulus lupulus L., Cannabaceae, é usada como sedativo e ansiolítico na medicina popular. O método de HPLC-DAD-ESI-MSn representa uma ferramenta poderosa para a análise de produtos naturais, desde que ela fornece o espectro de UV e informações estruturais sobre os constituintes da mistura. O objetivo deste trabalho foi o de caracterizar os constituintes encontrados no extrato hidroalcoólico. Os constituintes 1-9 foram tentativamente caracterizados através do UV/DAD e ionização por electrospray (MS/MS) depois da separação usando fase reversa, tempo de retenção e dados da literatura. Os principais compostos fenólicos (baseados na área dos picos) foram caracterizados como ácido hulupínico (9), coulupona (8), dois alfa-ácidos amargos oxidados (principais constituintes), um deles sendo um derivado da coumulinona oxidada (5) e o outro um derivado da humulinona oxidada (7), junto com uma procianidina B (3) e os flavonoides rutina (4) e o canferol-7-O-rutinosídeo (6). Esta planta conhecida devido às suas propriedades ansiolíticas e por ser um componente da cerveja, mostrou derivados oxidados de alfa-ácidos, como principais constituintes do extrato hidroalcoólico.

Humulus lupulus L., Cannabaceae, is commonly used as light sedative and anxiolytics in folk medicine. HPLC-DAD-ESI-MSn represents a powerful tool for the analysis of natural products, since it can simultaneously provide a UV chromatogram and significant structural information about compounds in complex mixture. The aim of this work was characterize the constituents present in hydroethanolic extract. Compounds 1-9 were tentatively characterized on the basis of UV, MS/MS, after reversed phase separation, retention time and literature data. The main phenolic compounds (based on peak area) were characterized as hulupinic acid (9), cohulupone (8), two oxidized hop alfa-bitter acids (principal constituents), one being a oxidized cohumulinone (5) and the other an oxidized humulinone (7) derivatives, together with a procyanidin dimer B (3), flavonoids rutin (4) and kaempferol-7-O-rutinoside (6). This plant known, due to anxiolytic property and beer flavoring, showed oxidized hop bitter acids, as principal constituents, in its hydroethanolic extract.

Quality Evaluation and Regional Analysis of Psoraleae Fructus by HPLC-DAD-MS/MS plus Chemometrics / 中草药·英文版

Objective To evaluate the quality of Psoralea corylifolia collected from 12 provinces of China.Methods An HPLCDAD-MS/MS method was used to identify,determine,and estimate 14 representative bioactive compounds in P.corylifolia.Then on the basis of the content data,the chemometrics method was used to differentiate 20 samples from different regions.Results The quality of P.corylifolia from 12 different provinces of China was evaluated by this method.Though the samples showed similar profiles,content of the detected markers varied significantly in different regions and batches.According to the results of the hierarchical cluster analysis and principle component analysis,it can be concluded that the samples from different origins could be clustered reasonably into two groups,as well as successfully distinguished.Conclusion A simple and reliable new method which used HPLC-DAD-MS/MS and chemometrics has been developed to characterize,classify,and control the quality of P.corylifolia.