An optimization strategy for charged aerosol detection to linearize the detector response in the multicomponent quantitative analysis of Qishen Yiqi dripping pills.

Charged aerosol detection, increasingly recognized for quantifying pharmaceutical compounds with weak ultraviolet absorption, is a universal detection technique for high-performance liquid chromatography (HPLC). Charged aerosol detection shows a non-linear response with increasing analyte concentration over a wide range, limiting its versatility in various analytical applications. In this work, a co-optimization strategy for power function value (PFV) and power laws was proposed and applied to broaden the linear range of the standard curve of saccharides in Qishen Yiqi dripping pills using the HPLC-charged aerosol detection (HPLC-CAD) method. Power function values for all analytes were optimized based on empirical models. Subsequently, the optimum power laws were investigated based on a preferred PFV. Additionally, various regression equations were evaluated to ensure the accuracy and precision of the results. With the optimized PFV and power law, the ordinary least squares model demonstrated a satisfactory fit. The optimal PFVs and power laws expanded the standard curve's linear range by 2.7 times compared to default settings, reducing model uncertainty. This paper presents a vital method for developing a multi-component quantitative HPLC-CAD approach without external data transformation outside the provided software, especially suitable for analytical applications of traditional Chinese medicine with significant quality differences.

Schisandra lignans ameliorate nonalcoholic steatohepatitis by regulating aberrant metabolism of phosphatidylethanolamines.

Nonalcoholic steatohepatitis (NASH) is a spectrum of chronic liver disease characterized by hepatic lipid metabolism disorder. Recent reports emphasized the contribution of triglyceride and diglyceride accumulation to NASH, while the other lipids associated with the NASH pathogenesis remained unexplored. The specific purpose of our study was to explore a novel pathogenesis and treatment strategy of NASH via profiling the metabolic characteristics of lipids. Herein, multi-omics techniques based on LC-Q-TOF/MS, LC-MS/MS and MS imaging were developed and used to screen the action targets related to NASH progress and treatment. A methionine and choline deficient (MCD) diet-induced mouse model of NASH was then constructed, and Schisandra lignans extract (SLE) was applied to alleviate hepatic damage by regulating the lipid metabolism-related enzymes CES2A and CYP4A14. Hepatic lipidomics indicated that MCD-diet led to aberrant accumulation of phosphatidylethanolamines (PEs), and SLE could significantly reduce the accumulation of intrahepatic PEs. Notably, exogenous PE (18:0/18:1) was proved to significantly aggravate the mitochondrial damage and hepatocyte apoptosis. Supplementing PE (18:0/18:1) also deteriorated the NASH progress by up regulating intrahepatic proinflammatory and fibrotic factors, while PE synthase inhibitor exerted a prominent hepatoprotective role. The current work provides new insights into the relationship between PE metabolism and the pathogenesis of NASH.

Separation of Flavonoids and Purification of Chlorogenic Acid from Bamboo Leaves Extraction Residues by Combination of Macroporous Resin and High-Speed Counter-Current Chromatography.

Flavonoids are major active small-molecule compounds in bamboo leaves, which can be easily obtained from the bamboo leaves extraction residues (BLER) after the polysaccharides extraction. Six macroporous resins with different properties were screened to prepare and enrich isoorientin (IOR), orientin (OR), vitexin (VI), and isovitexin (IVI) from BLER, and the XAD-7HP resin with the best adsorption and desorption performance was selected for further evaluation. Based on the static adsorption experiments, the experimental results showed that the adsorption isotherm fitted well with the Langmuir isotherm model, and the adsorption process was better explained by the pseudo-second-order kinetic model. After the dynamic trial of resin column chromatography, 20 bed volume (BV) of upload sample and 60% ethanol as eluting solvent was used in a lab scale-up separation, and the results demonstrated that the content of four flavonoids could be increased by 4.5-fold, with recoveries between 72.86 and 88.21%. In addition, chlorogenic acid (CA) with purity of 95.1% was obtained in water-eluted parts during dynamic resin separation and further purified by high-speed countercurrent chromatography (HSCCC). In conclusion, this rapid and efficient method can provide a reference to utilize BLER to produce high-value-added food and pharmaceutical products.

Insights into Q-markers and molecular mechanism of Sanguisorba saponins in treating ulcerative colitis based on lipid metabolism regulation.

BACKGROUND: Sanguisorba saponin extract (SSE) is the main active part of Sanguisorba officinalis with various pharmacological activities such as anti-inflammatory, anti-bacterial and anti-oxidant. However, its therapeutic role and underlying mechanisms for ulcerative colitis (UC) still need to be elucidated. PURPOSE: This study aims to explore the therapeutic effect, effectiveness-material basis-quality markers (Q-markers) and prospective mechanism of function of SSE on UC. METHODS: Fresh 2.5% dextran sulfate sodium salt (DSS) solution was placed in drinking bottles for 7 days to induce a mouse model of UC. SSE and sulfasalazine (SASP) were supplemented to mice by gavage for consecutive 7 days to investigate the therapeutic role of SSE on UC. Mouse monocyte macrophages (RAW264.7) and human normal colonic epithelial (NCM460) cells were treated with LPS to induce inflammatory responses, followed by pharmacodynamic examination with different concentrations of SSE. Hematoxylin-eosin (HE) and Alcian blue staining were conducted to evaluate the pathological damage of mice colon. Lipidomic technology was conducted to explore the differential lipids closely related to the disease process of UC. Quantitative PCR analysis, immunohistochemistry and ELISA kit were used to measure the expression levels of the corresponding proteins and pro-inflammatory factors. RESULTS: SSE treatment could effectively reduce the elevated expressions of pro-inflammatory factors in RAW264.7 and NCM460 cells due to LPS stimulation. Intragastric administration of SSE was found to significantly alleviate the symptoms of DSS-induced colon injury and low-polar saponins in SSE. Low polarity saponins, especially ZYS-II, were proved to be the main active substances of SSE in treating UC. In addition, SSE could significantly ameliorate the aberrant lipid metabolism in UC mice. The role of phosphatidylcholine (PC)34:1 in the UC pathogenesis has been fully verified in our previous studies. Herein, SSE-dosing effectively reversed the metabolic disorder of PCs in UC mice, and increased the PC34:1 level to normal via up-regulating the expression of phosphocholine cytidylyltransferase (PCYT1α). CONCLUSION: Our data innovatively revealed that SSE could significantly alleviate the symptoms of UC by reversing the disorder of PC metabolism induced by DSS modeling. SSE was proved for the first time to be a promising and effective candidate for UC treatment.

Establishing a chromatographic fingerprint using tandem UV/charged aerosol detection and similarity analysis for Shengmai capsule: A novel method for natural product quality control.

INTRODUCTION: Shengmai San, a well-known traditional Chinese medicine formula, is used to treat coronary heart diseases and myocardial infarction. The complex composition and complicated mechanism of the Shengmai preparations bring a significant challenge in the development of a suitable quality control method. OBJECTIVES: This work aims to establish a chromatographic fingerprinting method and propose a weighting algorithm for application in fingerprint similarity analysis to ensure consistent quality of the Shengmai capsule. METHODOLOGY: A chromatographic fingerprint method was established using tandem UV/charged aerosol detection (CAD) for Shengmai capsule quality control. After method verification, the developed method was applied to analyze 15 batches of the samples. Then a weighting algorithm of the fingerprint peak was proposed and used for the fingerprint similarity analysis. RESULTS: An HPLC-UV/CAD fingerprint method was successfully developed for the Shengmai capsules. Chromatographic conditions of the HPLC-UV/CAD method were optimized with a definitive screening design, and the optimized ranges of operating parameters were obtained with a Monte Carlo simulation method. The combined use of the proposed weighting algorithm and similarity analysis on fingerprint data improves the sensitivity of distinguishing batch-to-batch quality differences. CONCLUSION: The developed HPLC-UV/CAD fingerprint method is robust, reliable, and efficient. The proposed weighting algorithm combined with similarity analysis is promising and meaningful for the quality consistency assessment of HPLC-UV/CAD fingerprints.

Development of an HPLC-MS method for the determination of four terpene trilactones in Ginkgo biloba leaf extract via quality by design.

Previously reported HPLC-evaporative light scattering detection methods for terpene trilactone determination in Ginkgo biloba leaf extract (EGBL) have complicated sample preparation steps and are time-consuming. Thus, in this work, an HPLC-MS method for the determination of terpene trilactones in EGBL was developed with a novel analytical quality by design approach to provide robust and simple measurements. For this purpose, analytical target profiles and systematic risk analyses were performed to identify potential critical method attributes and critical method parameters. After screening experiments, a Box-Behnken design approach was utilized to investigate the relationships between critical method attributes and critical method parameters. A hypercube design space obtained by a Monte Carlo method was used for choosing the analytical control strategy. Then, verification experiments were performed within the design space, and the models were found to be accurate. After that, the optimized method was verified and successfully used for quality control analysis of EGBL from different manufacturers, and the results were almost the same as those determined by HPLC-evaporative light scattering detection. To our knowledge, this is the first study to establish a robust HPLC-MS method for determination of terpene trilactones in EGBL based on a novel analytical quality by design concept, which can improve the quality control of commercial EGBL.

Establishment and validation of the quantitative analysis of multi-components by single marker for the quality control of Qishen Yiqi dripping pills by high-performance liquid chromatography with charged aerosol detection.

INTRODUCTION: Charged aerosol detection (CAD) has the merits of high sensitivity, high universality and response uniformity. The strategy that combines the quantitative analysis of multi-components by single marker (QAMS) with CAD has certain advantages for the quantification of multi-components. However, relevant research was limited. OBJECTIVES: To comprehensively investigate the crucial factors that affect the performance of the HPLC-CAD-QAMS approach and validate the credibility and feasibility of the method. METHODOLOGY: Multiple components of Qishen Yiqi dripping pills (QSYQ) were assayed using the high-performance liquid chromatography (HPLC)-CAD-QAMS approach. Some factors that affect the sensitivity and accuracy of the approach were sufficiently studied. After the method verification, principal component analysis (PCA) was applied to evaluate the quality consistency of three types of samples: normal samples, expired samples and negative samples. RESULTS: A HPLC-CAD-QAMS method was successfully developed for the multi-component determination of QSYQ. First, chromatographic conditions were optimised by a definitive screening design, and the optimised ranges of operating parameters were obtained with a Monte Carlo simulation method. Next, a new method to select the internal reference standards was successfully introduced based on the heatmap of Pearson correlation coefficients of the response factors. Then, the multi-point method was selected to calculate the relative correction factors, and a robustness test was conducted with Plackett-Burman design. Finally, the PCA was proved to be effective for the quality consistency evaluation of different samples. CONCLUSION: The developed HPLC-CAD-QAMS method can be a reliable and superior means for the multi-component quantitative analysis of QSYQ.

The efficacy and safety of warming acupuncture and moxibustion on rheumatoid arthritis: A protocol for a systematic review and meta-analysis.

BACKGROUND: Rheumatoid arthritis (RA), as an autoimmune disease, can eventually lead to joint deformity and loss of function, seriously reduce the quality of life of patients and increase economic burden. As a traditional Chinese therapy, warming acupuncture and moxibustion is safe, economical, and has few side effects. At present, some studies have shown that warming acupuncture and moxibustion has a certain effect on RA, but there is no evidence of evidence-based medicine. The purpose of this study was to evaluate the efficacy and safety of warming acupuncture and moxibustion in the treatment of rheumatoid arthritis. METHOD: Randomized controlled trials of warming acupuncture and moxibustion treating RA will be searched in the databases including PubMed, EMBASE, the Cochrane library, Web of science, China National Knowledge Infrastructure (CNKI), WanFang, the Chongqing VIP Chinese Science and Technology Periodical Database (VIP), and China biomedical literature database (CBM) from inception to July, 2020. In addition, Baidu, Google Scholar, International Clinical Trials Registry Platform, and Chinese Clinical Trials Registry will be searched to obtain the gray literature and relevant data that have not yet been published. Two qualified researchers will extract data and assess the risk of bias from included studies independently. Statistical analysis is performed in RevMan 5.3 software. RESULTS: The primary outcome is symptom evaluation including morning stiffness, pain, and joint swelling. The number of joints affected by RA, Rheumatoid factor (RF), erythrocyte sedimentation rate (ESR), C reactive protein (CRP), anti-cyclic peptide containing citrulline (A-CCP), and adverse effects, will be evaluated as secondary outcomes. CONCLUSIONS: This study will compare the efficacy and safety of warming acupuncture and moxibustion with common acupuncture in the treatment of RA, providing reliable evidence for clinical application. OSF REGISTRATION NUMBER: DOI 10.17605/OSF.IO/C8RY9.

Comparative absorption kinetics of seven active ingredients of Eucommia ulmoides extracts by intestinal in situ circulatory perfusion in normal and spontaneous hypertensive rats.

Eucommia ulmoides Oliv. (E. ulmoides) is a valuable and nourishing medicinal herb in China that has been used in the treatment of hypertension. Given the fact that most traditional Chinese medicine is mainly used to treat disease, investigating the pharmacokinetics of traditional Chinese medicines in the pathological state is more useful than that in the normal state. However, the differences in the absorption kinetics of active ingredients of E. ulmoides extract between pathological and physiological conditions have not been reported. Therefore, in this study, the rat intestinal in situ circulatory perfusion model was used to investigate the differences in absorption kinetics of seven active ingredients of E. ulmoides extract in normal and spontaneously hypertensive rats, namely, genipinic acid, protocatechuic acid, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, (+)-pinoresinol di-O-ß-D-glucopyranoside and (+)-pinoresinol 4'-O-ß-D-glucopyranoside. Our results indicate that the pathological state of spontaneous hypertension may change the absorption of active components of E. ulmoides extracts, and these findings may provide a reference for improving the rational use of E. ulmoides in the clinic.

In-situ fabrication of Ag/P-g-C3N4 composites with enhanced photocatalytic activity for sulfamethoxazole degradation.

A series of Ag/P-g-C3N4 composites with different Ag content were synthesized for the first time by thermal polymerization combined with photo-deposition method. The composites were characterized by X-ray powder diffraction, field emission scanning electron microscope coupled with energy-dispersive X-ray spectroscopy, transmission electron microscopy, Fourier transform infrared spectroscopy, ultraviolet-visible diffuse reflectance spectra, N2 absorption-desorption and X-ray photoelectron spectroscopy. Ag was successfully dispersed on the surface of P-g-C3N4. The photocatalytic performance of P-g-C3N4 and Ag/P-g-C3N4 was evaluated by degrading sulfamethoxazole (SMX) under visible light irradiation. In the presence of 5% Ag/P-g-C3N4, 100% of SMX was degraded within 20 min. The enhanced photocatalytic activity of Ag/P-g-C3N4 was attributed to the surface plasmon resonance effect of metallic Ag and Schottky barrier formed on the interface between Ag and P-g-C3N4, which could speed up the generation rate of electrons and holes and inhibit the recombination of photogenerated electron-hole pairs. The radical quenching tests indicated that holes and superoxide radicals were the dominant active species involved in SMX degradation. The synthesized materials maintained high catalytic activity after five cycle runs. The concentration and the intermediates during the degradation process were determined by LC-MS/MS, and the tentative degradation pathways of SMX in photocatalytic system were proposed.

[Absorption of Inula cappa extract based on everted intestinal sac method].

To investigate the absorptive characteristics of Inula cappa extract based on the rat everted intestinal sac method in vitro. Nine representative ingredients in I. cappa extract were selected as the study objects. An UPLC-MS/MS method was established to determine and detect their cumulative absorption amount for expounding the absorptive characteristics of ingredients in different intestinal sections. According to the results， the transport mechanism of 8 compounds showed passive diffusion by the reverted gut sac method. And scopolin was actively transported in the intestine. The best absorption site of chlorogenic acid was duodenum. The best absorption site of cryptochlorogenic acid， 1，3-O-dicaffeoylquinic acid， luteolin-7-glucoside and 3，4-O-dicaffeoylquinic acid were jejunum. The best absorption site of neochlorogenic acid， scopolin， 4，5-O-dicaffeoylquinic acid and 3，5-O-dicaffeoylquinic acid was ileum. The absorption of all the compounds was affected by pH and bile. All of the nine ingredients in I. cappa extract could be absorbed in intestines， but with differences in the absorption rate， the best absorptive site and mechanism， indicating that the intestinal absorption of I. cappa extract was selective.

[Metabolism of Inula cappa extract by rat intestinal bacteria in vitro].

To investigate the metabolism of major components in Inula cappa by rat intestinal bacteria in vitro. I. cappa extract was incubated for 24 h with rat intestinal bacteria under anaerobic environment. After the samples were precipitated by n-butanol, UPLC-Q-TOF-MS/MS was applied for the qualitative analysis of the metabolites, combined with data software such as Metabolite Tools, Data Analysis and so on. The potential metabolites in rat intestinal bacteria were analyzed by comparing the total ion current of the test samples and blank samples and analyzing the quasi-molecular ion and fragment ion of all chromatograms. The results injected that fourteen metabolites were detected in rat intestinal flora. Various types of metabolic reactions happen to caffeoylquinic acid in intestinal flora, including isomerization, hydrolyzation, there were also methylation, hydrogenation and acetylation of caffeic acid. At the same time, a methylate of dicaffeoylquinic acid was also detected. Presumably, caffeoylquinic acids were gradually transformed into more hydrophobic metabolites with smaller molecular mass, which were better absorbed by the intestinal tract.

In vivo and in vitro antiviral activity of hyperoside extracted from Abelmoschus manihot (L) medik.

AIM: To assess the anti-hepatitis B virus (HBV) effect of hyperoside extracted from Abelmoschus manihot (L) medik. METHODS: The human hepatoma Hep G2.2.15 cell culture system and duck hepatitis B virus (DHBV) infection model were used as in vivo and in vitro models to evaluate the anti-HBV effects. RESULTS: In the cell model, the 50% toxic concentration of hyperoside was 0.115 g/L; the maximum nontoxic concentration was 0.05 g/L. On the maximum nontoxic concentrations, the inhibition rates of hyperoside on HBeAg and HBsAg in the 2.2.15 cells were 86.41% and 82.27% on d 8, respectively. In the DHBV infection model, the DHBV-DNA levels decreased significantly in the treatment of 0.05 g x kg(-1 ) x d(-1 ) and 0.10 g x kg(-1) x d(-1) dosage groups of hyperoside (P<0.01). The inhibition of the peak of viremia was at the maximum at the dose of 0.10 g x kg(-1 ) x d(-1) and reached 60.79% on d 10 and 69.78% on d 13, respectively. CONCLUSION: These results suggested that hyperoside is a strong inhibitor of HBsAg and HBeAg secretion in 2.2.15 cells and DHBV-DNA levels in the HBV-infected duck model.