The quality evaluation system of a famous traditional Chinese medicine Huaganjian decoction was established.

Huaganjian decoction (HGJD) has been widely used clinically to treat liver injuries and gastritis. However, the quality evaluation system for HGJD is not perfect. In this study, paeoniflorin, hesperidin, geniposide, naringin, and quercetin were employed as quality markers. The quantitative analysis of these five components in HGJD was conducted using a high-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry method. This method underwent validation for linearity, precision, accuracy, repeatability, and recovery. In summary, a reliable quantitative method was successfully employed to establish a comprehensive quality evaluation of HGJD.

Analysis of phytochemical components of Tibetan medicine Pedicularis flava and Pedicularis muscicola by GC-MS and UHPLC-TOF-MS.

Traditional medicine, 'LuRu', is a commonly used Tibetan medicine for clearing away heat and detoxifying. Dried products of Pedicularis flava and Pedicularis muscicola are often used as 'LuRu' in the market. This study aims to compare the chemical constituents of P. flava and P. muscicola using GC-MS and UPLC-TOF-MS, and confirm which plant species is more suitable to be used as 'LuRu'. A total of 46 and 68 compounds were identified from the volatile and non-volatile components, respectively. Out of these, 17 and 37 volatile and non-volatile components, respectively, had pharmacological activities. P. flava showed a higher content of the same active components than P. muscicola. Good biological activities are only observed in the unique components in P. flava, and not in P. muscicola. The two herbs should not be mixed in clinical medication. Our study shows that P. flava is better suited as a high-quality herb for the Tibetan medicine, 'LuRu'.

Volatile components of Tibetan Pedicularis flava and Pedicularis muscicola were analysed for the first timewiooi.Non-volatile components of Tibetan Pedicularis flava and Pedicularis muscicola were analysed for the first time.Differences in chemical composition and content between Pedicularis flava and Pedicularis muscicola were studied.

Comprehensive chemical profiling of the flowers of Citrus aurantium L. var. amara Engl. and uncovering the active ingredients of lipid lowering.

The flowers of Citrus aurantium L. var. amara Engl. (FCAVA) is popularly consumed as an edible tea for anti-hyperlipidemia. But the active ingredients are not fully clear. In this study, ultra-high performance liquid chromatography coupled to quadrupole time of flight tandem mass spectrometry (UHPLC-QTOF-MS/MS) with diagnostic product ions and neutral loss filtering strategy were successfully used for comprehensive characterization of chemical components in FCAVA. A total of 228 constituents, including 46 organic acids, 12 coumarins and 170 flavonoids, were tentatively characterized (30 confirmed with reference standards). Among them, nineteen flavonoids in 70 batches of FCAVA from different geographical origins were quantified by UHPLC tandem triple quadrupole mass spectrometry (QQQ-MS), which displayed satisfactory linearity, sensitivity, precision, accuracy, and stability. According to analytical results, the distribution of nineteen flavonoids in different geographical origins of FCAVA was clarified. In addition, the effect on LDL uptake of twenty-five flavonoids was investigated in HepG2 cell. It was found that the acacetin, diosmetin and rutin dose-dependently enhanced LDL uptake in HepG2 cells comparing to control. Furthermore, in a hyperlipidemia C57BL/6J mice model, administration of acacetin, diosmetin and rutin (30 mg/kg/d, intragastric, for three weeks) significantly decreased the levels of total cholesterol (TC), triglyceride (TG) and low density lipoprotein cholesterol (LDL-C) in plasma, respectively. Overall, these findings indicated the potential of FCAVA in the development of functional food or medicine for the prevention and treatment of hyperlipidemia, which could be considered for the improvement of quality standardization of FCAVA.

Esculetin as bioactive marker: towards a rational scientific approach for the treatment of hyperuricemia using Traditional Chinese Medicine

Traditional Chinese Herbal Medicine has been used to prevent and cure disease in China for thousands of years and has gained global interest in recent decades. The Erding formula is a Chinese Pharmacopeia (ChP)-listed herbal preparation used for treating sore throat, carbuncles and boils. Esculetin is a ChP quality control (QC) marker for these indications. A previous study found that a new indication, hyperuricemia, can be added to the Erding formula. Therefore, this study aimed to evaluate whether the traditionally used marker, esculetin, still has bioactivity for hyperuricemia, which is substantially different from the original indications. The study analyzed the quantity of esculetin by high-performance liquid chromatography, assessed the therapeutic effect of esculetin using animal model, and then characterized esculetin and its metabolites in serum via ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry. The results showed that the esculetin content in the aqueous Erding extract was 0.26±0.05% (w/w). Both the Erding extract and esculetin significantly reduced uric acid levels. Six metabolites of esculetin were identified in mice serum. This study revealed a rational scientific approach to prove esculetin is a reliable bioactive and QC marker for Erding formula in hyperuricemia treatment which contributed to ensure product quality and therapeutic efficacy.

Constituents and Anti-Hyperuricemia Mechanism of Traditional Chinese Herbal Formulae Erding Granule.

Erding granule (EDG) is a traditional Chinese medicine that has recently been identified as having anti-hypouricemic effects. However, the active components and underlying mechanism for this new indication have not been elucidated. Therefore, we compared the effects of different EDG extracts (water, 50% ethanol and 95% ethanol) on serum uric acid concentrations in the hyperuricemia model mouse. We also analyzed the constituents of different extracts by ultra-high performance liquid chromatography combined with electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS/MS) to observe the variation between the active and inactive products. Extract activity and target site were evaluated by assessing uric acid- and inflammation-suppressing effects along with evaluating ability to regulate the uric acid transporter. The results showed that the 50% ethanol extract (EDG-50) had an obvious serum uric acid concentration lowering effect compared with water (EDG-S) and the 95% ethanol extract (EDG-95). UHPLC-Q-TOF-MS/MS analysis showed that EDG-50 was compositionally different to EDG-S and EDG-95. EDG-50 showed dose-dependent effects on reducing uric acid, suppressing inflammation and regulating uric acid transporters. Moreover, western blot analysis showed that EDG-50 down-regulated GLUT9 and URAT1 expression, and up-regulated OAT1 expression. Therefore, our findings enable the preliminarily conclusion that EDG-50 lowers serum uric acid concentrations, mainly by down-regulating the expression of GLUT9 and URAT1 proteins and up-regulating the expression of OAT1 proteins. This provides a research basis for clinical use of EDG as an anti-hyperuricemic agent.

Tissue Distribution and Anti-Lung Cancer Effect of 10-Hydroxycamptothecin Combined with Platycodonis Radix and Glycyrrhizae Radix ET Rhizoma.

10-Hydroxycamptothecin (HCPT) is a broad-spectrum chemotherapeutic drug, although its side effects and multidrug resistance (MDR) limit its clinical application. A range of drug delivery systems have been utilized to overcome its shortcomings and maintain its therapeutic efficacy, however the use of the transport effect of traditional Chinese medicines (TCMs) to improve the distribution of chemotherapeutic drugs has not been widely reported. Platycodonis Radix (JG) and Glycyrrhizae Radix ET Rhizoma (GC) are common TCMs in clinics and are often combined as drug pairs to act as "transport agents". In the present study, the effect of JG and GC (JGGC) on the distribution of HCPT in tissues and its antitumor efficacy after being combined as a therapy were investigated, for which ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was used. Furthermore, the effect on the protein expression of multidrug resistance proteins (P-gp and LRP), and the immunomodulatory and synergistic antiapoptotic effect on Lewis lung cancer-bearing C57BL/6J mice were also evaluated. The results demonstrate that JGGC significantly increased the area under the concentration time curve (AUC) and mean residence time (MRT) and reduced the clearance rate (CL) of HCPT. In addition, the combined use of JGGC decreased the levels of LRP, P-gp and Bcl-2/Bax when treated with HCPT. JGGC also significantly elevated the levels of RBCs, PLTs, HGB, IL-2, and IFN-γ, and decreased IL-10 levels. In summary, an increased concentration of HCPT in tissues was observed when it was combined with JGGC through inhibition of efflux protein, with a synergistic enhancement of the anticancer effect observed through promotion of apoptosis and immunity due to a reversion of the Th1/Th2 shift. Our findings provide a reference for the feasibility of combining JGGC with chemotherapy drugs in clinical applications.

Study on anti-hyperuricemia effects and active ingredients of traditional Tibetan medicine TongFengTangSan (TFTS) by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry.

TongFengTangSan (TFTS), a traditional Tibetan medicine comprising of Tinospora sinensis (TS), Terminalia chebula Retz (TC) and Trogopterori faeces (TF), is used to treat joint diseases like gout, gout arthritis, swelling, pain etc. Despite the significant therapeutic effects of TFTS, its pharmacological components have not been analyzed so far. Therefore, the chemical composition of the effective part of TFTS was analyzed by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS/MS). The results show that the ethanol extract (EE) of TFTS was more effective in reducing the serum uric acid (SUA) and XOD (Serum and Liver) levels in a hyperuricemic rats model compared to the TFTS raw powder (RP). UHPLC-Q-TOF-MS/MS identified a total of 106 compounds in the positive and negative ion mode, of which 87 were from TC, 13 from TS and 6 from TF. In addition, 106 compounds contained 57 tannins, 6 triterpenoids, 10 alkaloids, 7 flavonoids, 22 organic acids and 4 phenylpropanoids. The preliminary results indicate that the EE of TFTS includes the active anti hyperuricemic substances. The present study first investigated the efficacy and the active components of TFTS in hyperuricemic treatment, and further summarized the diagnostic ion and neutral loss patterns of MS/MS cracking of tannic compounds. These findings lay the foundation for the further study and clinical application of TFTS.

Identification and Quantification Analysis on the Chemical Constituents from Traditional Mongolian Medicine Flos Scabiosae Using UHPLC-DAD-Q-TOF-MS Combined with UHPLC-QqQ-MS.

In this study, a systematic method was established for the qualitative and quantitative analysis of the major constituents in Flos Scabiosae (FS). First, Ultra-high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) was developed for the identification of the multi-constituents in FS. A total of 48 compounds (9 phenolic acids, 24 flavonoids, 8 iridoids and 7 others) were unambiguously or tentatively identified, including 25 compounds (flavonoids, phenolic acids) identified in FS for the first time. Second, ultra-high performance liquid chromatography combined with triple quadrupole mass spectrometry (UHPLC-MS-MS) was developed for the quantitative analysis of 10 phenolic compounds. Ten compounds, either with high contents or strong bioactivities, were chosen as markers. This analytical method was validated through intra- and inter-day precision, repeatability and stability, with respective relative standard deviations <4.43, <8.64, <4.60 and <3.65%, respectively. The limits of detection and quantification were <1.09 and <16.96 ng/mL, respectively. The overall recoveries ranged from 96.47 to 103.94% .Then the validated method was applied to determine 10 batches of FS. The results indicated that the new method can be applied for the qualitative and quantitative analysis of FS.

[Quantitative analysis of phenolic acids in ainsliaea fragrans from different habitats].

OBJECTIVE: The contents of major phenolic acids in Ainsliaea fragrans from different habitats were determined,in order to improve quality standards of Ainsliaea fragrans and provide reference for optimization its daodi habitat. METHODS: Separation and determination of two kinds of phenolic acids, caffeoylquinic acids (CQA)and dicaffeoylquinic acid (DCQA), in Ainsliaea fragrans were carried out by RP-HPLC with Cosmosil RP-C18 column (250 mm x 4.6 mm, 5 µm), acetonitrile-0.1% formic acid solution as mobile phase at the flow rate of 1.0 mL/min, detection wavelength at 328 nm and column temperature at 35 °C. RESULTS: CQA and DCQA were uneven in Ainsliaea fragrans from different habitats. In the sample from Wuyuan, Jiujiang and Shangrao in Jiangxi, the contents of phenolic acids were higher than those from other habitats significantly. The sample from Guangxi habitat had the lowest phenolic acids content. And the chlorogenic acid and 3,5-DCQAs' percentage were higher than other phenolic acids. CONCLUSION: This method is accurate and easy with good repeatability, which can be used in determination of eight phenolic acids in Ainsliaea fragrans from different habitats. Ainsliaea fragrans in Jiangxi Province has good qulity.

High-performance liquid chromatography/tandem mass spectrometry for simultaneous determination of four dicaffeoylquinic acids in rat plasma.

A specific and reliable HPLC-MS/MS method was developed and validated for the simultaneous determination of four dicaffeoylquinic acids (DCQA): 3,4-DCQA, 1,5-DCQA, 3,5-DCQA and 4,5-DCQA. The analytes were separated on a C18 column (150 mm × 2.1 mm, 1.8 µm) and a triple-quadrupole mass spectrometry equipped with an electrospray ionization (ESI) source was applied for detection. The plasma sample was prepared by a liquid-liquid extraction method and the recovery for the four analytes was around 80%. The calibration curves were linear over a concentration range of 10.6-1060.0 ng/mL for 3,4-DCQA, 19.2-1920.0 ng/mL for 1,5-DCQA, 14.0-2900.0 ng/mL for 3,5-DCQA, 9.7-970.0 ng/mL for 4,5-DCQA. The intra-day and inter-day precision was less than 15% and the relative error (RE) were all within ±15%. The validated method was successfully applied to a pharmacokinetics study in rats after oral administration of the extracts of Ainsliaea fragrans cham (a traditional Chinese herb).

Simultaneous quantification of polymethoxylated flavones and coumarins in Fructus aurantii and Fructus aurantii immaturus using HPLC-ESI-MS/MS.

The major lipid-soluble constituents in Fructus aurantii (zhiqiao) and Fructus aurantii immaturus (zhishi) are polymethoxylated flavones (PMFs) and coumarins. In the present study, a high-performance liquid chromatography with electrospray ionization tandem mass spectrometry method was developed to quantify PMFs (nobiletin, tangeretin, 5-hydroxy-6,7,8,4'-tetramethoxyflavone, and natsudaidai) and coumarins (marmin, meranzin hydrate, and auraptene) simultaneously. PMFs and coumarins were detected by electrospray ionization tandem mass spectrometry in positive ion mode and quantified with multiple reaction monitor. Samples were separated on a Diamonsil C18 (150 mm × 4.6 mm, 5 µm) column using acetonitrile and formic acid-water solution as a mobile phase in gradient mode with a flow rate at 0.5 mL/min. All calibration curves showed good linearity (r² > 0.9977) within the test ranges. Variations of the intraday and interday precisions were less than 4.07%. The recoveries of the components were within the range of 95.79%-105.04% and the relative standard deviations were less than 3.82%. The method developed was validated with acceptable accuracy, precision, and extraction recoveries and can be applied for the identification and quantification of four PMFs and three coumarins in citrus herbs.

[Study on antibacterial active components from Viola yedoensis].

OBJECTIVE: Study on the antibacterial activity of Viola yedoensis and the antibacterial active compounds. METHOD: The chemical compositions were isolated by means of solvent extraction, column chromatography on silica gel, sephadex LH-20 and crystallization. The antibacterial activities were tested by Neo-Sensitab disk-diffusion method, nephelometric analysis and plating method. RESULT: One new compound (4) along with three known compounds were isolated from this plant for the first time and were identified as aesculetin (1), 6,7-dimethoxycoumarin (2), scopoletin (3) and 5-methoxy-7-hydroxymethylcoumarin (4), respectively. All the compounds showed antibacterial and antibactericidal activities at varying degree on Streptococcus Aureas, S. agalactiae, S. uberis, S. dysgalactiae, E. coli and Salmonella, of which 1 was most active with 0.031- 0.313 g x L(-1) of minimal inhibitory concentrations (MIC) and 0.313 - 0.625 g x L(-1) of minimal bactericidal concentrations (MBC). CONCLUSION: Viola yedoensis has a broad spectrum of antibacterial activity on animal pathogenic bacteria, and coumarins may be the main antibacterial activity ingredients.