High-performance Liquid Chromatography Fingerprint Research Articles

A strategy for simple multi-component quantification and comprehensive quality consistency evaluation using fingerprinting technology in the case of Bazhen Yimu pill

Bazhen Yimu Pill is developed on the basis of the famous “Eight Treasures Soup”, which has the efficacy of treating weakness of spleen and stomach, deficiency of qi and blood, and other conditions. Since the drug consists of nine Traditional Chinese medicines (TCM) with complex herbal components, quality control is difficult, so it is necessary to explore and innovate effective quality control methods. In this study, we firstly established a high-performance liquid chromatographic fingerprint for the markers (gallic acid, chlorogenic acid, ferulic acid, paeoniflorin and leonurine) in BZYMP using a reliable single-linear multiple-labeling (MAML) method for experimental accurate and rapid multi-component quantification. Subsequently, the systematic quantitative fingerprinting method (SQFM) was used to evaluate the HPLC fingerprints (HPLC-FP), ultraviolet quantum fingerprints (UV-QFP), and differential scanning quantum fingerprints (DSC-QFP) to obtain rich qualitative and quantitative fingerprint information. In order to improve the objectivity and rationality of the comprehensive evaluation results, the CVWM algorithm was used to perform fusion integration of HPLC-FP, UV-QFP and DSC-QFP. The results show that the quality grades after fusion integration are categorized into 1–8, and the fusion integration is more capable of evaluating the overall quality of BAYMP from multiple perspectives and levels than individual experiments. The quantitative analysis results of MAML are more accurate compared to SCM and QAMS. In this study, BZYMP was analyzed using multidimensional fingerprinting to provide a comprehensive and reliable method for quality consistency control of TCM and its preparations.

Identifying the active antioxidant ingredients in Arisaema Cum Bile aqueous extract on the basis of spectrum-effect relationships

At present, the research on Arisaema Cum Bile mainly focuses on the analysis of its components and changes in pharmacological effects. The chemical composition of the water extract, the material basis and mechanism of its anti-oxidation and anti-febrile convulsion are still unclear. In this study, fingerprint-pharmacological relationships were used to determine the pharmacodynamic and pharmacological material basis of the antioxodant effects of Arisaema Cum Bile aqueous extract. MethodsThe High Performance Liquid Chromatography (HPLC) fingerprint of the aqueous extract of Arisaema Cum Bile was established by determining the common peaks, recording the peak areas and evaluating the similarities between different batches. Moreover, the antioxidant capacity of each batch of processed Arisaema Cum Bile extracts was determined, and spectrum-effect relationship analysis and screening were conducted on the basis of the fingerprint. The antioxidant activities of the Arisaema Cum Bile aqueous extract ingredients were verified by evaluating the isolated compounds. ResultsThe HPLC fingerprints of 15 batches of aqueous extracts of Arisaema Cum Bile aqueous extracts were established via ultraviolet (UV) and evaporative light scattering detection methods. There was a total of 22 common peaks, 11 of which were identified as uracil, hypoxanthine, xanthine, uridine, guanosine, adenosine, shaftoside, isoschaftoside, taurohyodeoxycholic acid, taurochenodeoxycholic acid and glycinechenodeoxycholic acid. Spectrum-effect relationship analysis revealed that shaftoside, isoschaftoside, taurochenodeoxycholic acid and glycochenodeoxycholic acid may be the active antioxidant ingredients in this extract, which was verified since all four ingredients displayed dose-dependent antioxidant effects when tested individually. ConclusionA fingerprint of the aqueous extract of Arisaema Cum Bile aqueous extract was established, the antioxidant capacities of the extracts were measured, and the active antioxidant ingredients were identified through spectrum-effect relationship analysis.

Study on HPLC fingerprint of Saposhnikovia divaricata lipophilic components and anti-aging effects

Saposhnikovia divaricata is commonly used in clinical practice as a heat-clearing and detoxifying traditional Chinese medicine. Current research on Saposhnikovia divaricata has mostly focused on chromones, while studies on its lipophilic parts remain scarce. In this study, samples of the lipophilic components of Saposhnikovia divaricata (SDL) from different regions in China were collected to establish an HPLC fingerprint. The quality of SDL was evaluated through chemical pattern recognition and multi-component content determination. Furthermore, using the Caenorhabditis elegans (C. elegans) model, the anti-aging ability of SDL was preliminarily studied. In the HPLC fingerprints of 11 batches of SDL, 19 common peaks were identified, and 4 components were characterized as follows: peak 14, linolenic acid (ALA); peak 15, docosahexaenoic acid (DHA); peak 16, linoleic acid (LA); peak 17, oleic acid (OA). The similarity of the fingerprint ranged from 0.704 to 0.929. The results indicate that the content of ALA, DHA, LA, and OA are 0.040–0.440 mg/g, 0.003–0.116 mg/g, 1.047–6.460 mg/g, and 0.040–0.294 mg/g, respectively. HCA analysis divided 11 batches of SDL into two categories, and PCA analysis extracted 6 principal components with a cumulative contribution rate of 85.91%. The anti-aging ability of the SDL was proved by measuring the lifespan, reproductive capacity, body length, mobility, lipofuscin content, and heat stress resistance of C. elegans. The combination of high-performance liquid chromatography fingerprint and chemical pattern recognition for multi-component content determination is a reliable, comprehensive, and promising evaluation of the quality of SDL. The confirmation of SDL’s anti-aging ability provides the possibility for the research of Saposhnikovia divaricata in the field of healthy food and cosmetics.

A strategy of Q-markers identification based on effect, property flavour material basis and rapid quantitative evaluation via near-infrared spectroscopy and chemometric methods for the quality control of Flos Trollii (FT)

Ethnopharmacological relevanceFlos Trollii (FT) is the dried flower of Trollius Chinensis Bunge of Ranunculaceae with the pharmacological properties of anti-inflammatory, antibacterial, antiviral, anti-oxidative. The herb FT is not only a traditional Chinese medicine (TCM) but also an extensively utilized ethnic medicine, employed by diverse ethnic groups including Mongolian, Tibetan, and Kazakh. Aim of studyFT was taken as an example to construct a strategy of quality markers (Q-markers) identification based on effect, property flavor material basis, and rapid quantitative evaluation using near-infrared (NIR) spectroscopy and chemometric methods of TCM. Materials and methodsInitially, the anti-inflammatory efficacy of FT from three places of origin was evaluated using the RAW264.7-cell inflammatory model, and the bitter property flavor was characterized using an electronic tongue. The high-performance liquid chromatography(HPLC) fingerprint of FT was generated, and the quality of FT from different origins was evaluated employing chemometrics. Next, potential anti-inflammatory and bitter property flavor compounds were screened utilizing a fingerprinting-effect relationship and fingerprinting-property flavor relationship model using partial least squares regression (PLSR). The Q-markers of the FT were confirmed based on the testability principle. Then, a swift, uncomplicated, and precise Q-marker content of the FT prediction model was developed by adopting NIR. ResultsThe main common fingerprinting peaks affecting FT's efficacy and property flavor were screened. Five of these compounds, 2″-O-beta-L-galactopyranosylorientin, orientin, vitexin, veratric acid, and isoquercitrin, characterized using HPLC and ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS), could be regarded as Q-markers of FT. Q-marker content of the FT prediction model developed adopting NIR spectroscopy was rapid and effective. ConclusionAccording to the strategy proposed in this study, a quantitative NIR spectroscopic method to identify Q-markers could be a tool to improve the QC efficiency of TCM.

Comprehensive Investigation of Homology and Heterogeneity Between Aurantii Fructus and Aurantii Fructus Immaturus Using Chemometrics Combined with Network Pharmacology.

Aurantii Fructus (AF) and Aurantii Fructus Immaturus (AFI) are distinct herbs outlined by the Chinese Pharmacopoeia. They are sourced from the same plant but harvested at different times, resulting in differences in efficacy. It is important to avoid mixing them clinically and to distinguish between the two. Furthermore, dissimilar cultivation conditions may cause variability in the quality of herbs, so it is vital to differentiate drugs from dissimilar origins. In this study, two plants, AF and AFI from different provinces, were comparatively analyzed based on High Performance Liquid Chromatography (HPLC) fingerprints and classified using chemometric methods. The results indicate that the two medicines can be clearly distinguished. Also, AF and AFI grown in different locations can be distinguished. Ten chemical markers were screened, and their variations were determined, including eriocitrin, narirutin, naringin, meranzin hydrate, naringenin, hesperidin, nobiletin, tangeretin, neohesperidin, and poncirin. Subsequent network pharmacology correlated the screened chemical components with the biological network of the organism. The material basis of the difference in efficacy of the two homologous herbs was explored from the perspective of changes in chemical composition. This study provides a reference for formulating quality evaluation standards for AF and AFI and lays a foundation for the efficacy-related quality research of the two.

Phytochemical screening, HPLC fingerprinting and in vitro assessment of therapeutic potentials of different apricot cultivars against diabetes, Alzheimer's disease and cancer

Plant-based natural compounds are widely used to treat various ailments owing to their readily availability and minimal adverse effects. This study aimed to perform qualitative and quantitative biochemical profiling and assess the in vitro anti-diabetic, anti-Alzheimer, and anti-cancer activities of various apricot (Prunus armeniaca) cultivars. High-performance liquid chromatography (HPLC) was utilized to determine the concentrations of bioactive compounds across 10 distinct apricot cultivars. Initial phytochemical screening revealed a significant content of secondary metabolites. Subsequently, methanolic extracts from these cultivars were evaluated for their therapeutic potential against several human cancer cell lines, including prostate cancer (PC-3), lung cancer (A-549), breast cancer (MCF-7), cervical cancer (HELA), and kidney cancer (HEK). Notably, the breast cancer cell line MCF-7 showed a pronounced inhibition rate post-treatment with the apricot extracts. Correlation analysis exhibited phenols are highly correlated with flavonoids (r = 0.92), DPPH (r = 0.95), and alpha-amylase (%) inhibition (r = 0.96), and showed a significant correlation with other parameters. Principal Component Analysis (PCA) revealed that PC1 explained 43.31 % of the variance, while PC2 explained 12.88 %, together explaining 80.033 % of the total variance. PC1 was identified as the dominant axis, indicating the primary pattern of variation among the variables. Hierarchical Cluster Analysis (HCA) divided the cultivars into 2 main clusters, with cluster 2 further subdivided into various sub-clusters and sub-sub-clusters. This analysis highlighted distinct genetic similarities and differences among the apricot cultivars. Among the tested cultivars, ‘Irani’ and ‘Tilton’ were found to contain the highest levels of bioactive constituents. This research marks the first comprehensive examination of the impacts of these two apricot cultivars. The findings from this study provide a robust scientific foundation for the future isolation and purification of therapeutic compounds, potentially leading to their application in pharmaceuticals or dietary supplements. This research contributes significantly to the understanding of the pharmacological properties of apricot cultivars and establishes a basis for further investigation into their clinical benefits.

Shugan Jiangzhi Decoction Alleviates Nonalcoholic Fatty Liver Disease (NAFLD) via Regulating AMPK/PPAR Signaling Pathway

Background: Non-alcoholic fatty liver disease (NAFLD) is considered the most common liver disease. Shugan Jiangzhi Decoction (SJD), a traditional Chinese medicine (TCM) formula which consists of six Chinese herbs, has been used for treating hyperlipidemia, obesity, and fatty liver in Guangdong Provincial Hospital of Traditional Chinese Medicine for over twenty years. Objective: This study aims to elucidate the chemical basis and the molecular mechanism of SJD against NAFLD. Methods: The main components of SJD were determined by High Performance Liquid Chromatography (HPLC). Then the high-fat diet (HFD)-induced NAFLD rat model was established. After treatment with different doses of SJD, the body weight of rats was measured weekly. On the last day of the experiments, the hepatic morphology, histopathology changes, and the serum lipid levels were detected. Then techniques of network pharmacology were employed to predict the anti-NAFLD mechanism of SJD. At last, the expression levels of proteins were measured by western blot to verify the mechanism. Results: Nine chemical constituents of SJD were identified from HPLC fingerprint spectrum. For the in vivo experiment, NAFLD rat model was constructed successfully by feeding high-fat diet (HFD) for 8 weeks. The following treatment with SJD for 6 weeks decreased the fatty droplet accumulation in the liver obviously. Meanwhile, the serum level of high-density lipoprotein cholesterol (HDL-c) was increased, while the levels of lowdensity lipoprotein cholesterol (LDL-c), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) were decreased after SJD treatment (p < 0.05). The results of network pharmacology indicated that SJD might improve NAFLD via regulating the AMPK/PPAR signaling pathway. Then, the western blot assay confirmed that SJD activated the AMPK/PPAR signaling pathway in the liver of rats. Conclusion: SJD improves HFD-induced NAFLD in rats via AMPK/PPAR signaling pathway. Thus, our study suggests that SJD can serve as a therapeutic agent for the prevention and treatment of NAFLD.

Characterization of the phytochemical constituents of methanol extract of <i>Spondias mombin</i> stem bark using high performance-liquid chromatography and gas chromatography mass spectroscopy

Spondias mombin is commonly used in traditional medicine to treat various ailments. This study was aimed to analyze the bioactive components of the methanol extract of S. mombin stem by High-performance liquid chromatography (HPLC) and Gas Chromatography-Mass Spectroscopy (GC-MS). Quantitative analysis of the extract showed that it had appreciable quantities of flavonoids, phenols, tannins, and proanthocyanidins. The HPLC fingerprints of the extract also revealed the presence of flavonoids, tannins, saponins, alkaloids, other phenols, and steroids. The most prevalent flavonoids were the flavones, and the alkaloids present in high concentrations were ribalinidine and linamarin. Some other phenolics, such as resveratrol, ellagic acid, chlorogenic and pyrogallic, were in high concentration. The GC-MS data detected bioactive compounds; notably among them were 9,12 -octadecadienoic acid (linoleic acid), n-hexadecanoic acid, and 9-oxo nonanoic acid. This study has established the presence of bioactive chemicals in the methanol extract of S. mombin stem bark, and these phytochemicals could be responsible for the plant’s medicinal properties.

Characterization and In vitro Release & In vivo Behavior Study of Self-Assembled Nano-Emulsion in XiaoYao Pill for Enhanced Drug Delivery.

Traditional Chinese medicine formulations often contain hydrophobic components with limited solubility and stability, leading to low oral bioavailability. Self-assembled nanoparticles (SANs) have shown promise in enhancing oral bioavailability of these components. However, whether un-decocted Chinese herbal pellets can generate SANs and the impact of SANs formed by multiple components on pharmacokinetic parameters remains unexplored. In this study, single-factor approach was employed to determine the optimal separation method of nano-emulsion phase of XiaoYao pill (N-XY). Morphological and particle size analyses confirmed the nanoscale nature of N-XY. High-performance liquid chromatography (HPLC) fingerprint analysis was conducted to compare the distribution of active ingredients among three different phases of XiaoYao pill (XY pill). In vitro release studies were performed to evaluate the release mechanism of four ingredients from N-XY. Additionally, in vivo pharmacokinetics and tissue distribution behaviors were investigated in rats. N-XY exhibited uniform and stable characteristics as a water-in-oil (O/W) nano-emulsion. Fingerprint analysis identified 25 characteristic peaks and 8 key ingredients in N-XY, with the highest peak areas. In vitro release studies showed a sustained release behavior of N-XY. The pharmacokinetics study showed that the ferulic acid of N-XY had a 1.37-fold higher AUC, 1.44-fold lower Vd/F, 1.39-fold lower CL/F, and a prolonged t1/2 than A-XY, indicating enhanced bioavailability due to reduced elimination. Furthermore, the tissue distribution revealed that the levels of paeoniflorin and ferulic acid from N-XY significantly increased in liver, spleen, lungs, uterus and ovaries, exhibiting targeting characteristics. This study comprehensively explored the formation, characterization, and pharmacokinetics of nano-emulsion in XY pill, introducing novel perspectives and initiating preliminary research on potential SANs in un-decocted traditional Chinese medicine formulations. It also emphasized the importance of enhancing pharmacokinetics of hydrophobic components in Chinese herbal formulations and laid the foundation for future nano-formulation research for XY pill.

A novel evaluation mode of Nao An Capsules using multi-wavelength fusion fingerprint combined with quantitative analysis of multi-components by a single marker method and antioxidant activity

Nao An Capsules (NAC) are utilized in the acute phase of cerebral thrombosis and are appropriate for patients in the recovery stage exhibiting symptoms of qi deficiency and blood stasis. Nevertheless, there has been a lack of recent research on the overall quality control of NAC. The objective of this study is to establish a comprehensive quality control strategy for NAC. This article presents a methodology based on the average linear quantitative fingerprinting method (ALQFM) in conjunction with multi-wavelength high performance liquid chromatography (HPLC) fusion fingerprinting, quantized Fourier transform infrared (FT-IR) spectroscopy and ultraviolet (UV) spectral techniques to comprehensively evaluate NAC from both qualitative and quantitative perspectives. Subsequently, quantitative analysis of multi-components by a single marker (QAMS) method and the Standard Curve Method (SCM) were used to quantitatively determine 10 quality markers (Q-markers) in 22 batches of samples, with the results indicating that QAMS is a reliable method for quantifying active ingredients. Chemometrics methods such as principal component analysis (PCA) and principal component analysis (HCA) were also used to analyze the quality of samples. To achieve a thorough and methodical evaluation, a combined assessment of the three fingerprints was executed. Subsequently, an in vitro antioxidant activity experiment was performed utilizing the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging technique. By employing a partial least squares (PLS) model, the correlation between HPLC fingerprint chromatography and antioxidant activity was investigated. The study found that out of the 59 common peaks in the five-wavelength fusion fingerprint spectra of NAC, 33 peaks exhibit antioxidant activity. In this study, multi-wavelength fusion HPLC, quantized FT-IR and UV were jointly used to effectively identify functional groups and fingerprint information in compounds. It can fully and quickly obtain the fingerprint information of the compounds in the sample. NAC multi-components quantification and overall quality evaluation were performed using ALQFM and QAMS and chemometrics methods. We have developed a new approach for comprehensive, rapid, and cost-effective quality control of traditional Chinese medicine (TCM) compound preparations.

Evaluating the impact of ecological factors on the quality and habitat distribution of Lonicera japonica Flos using HPLC and the MaxEnt model.

The quality of traditional Chinese medicine is based on the content of their secondary metabolites, which vary with habitat adaptation and ecological factors. This study focuses on Lonicera japonica Flos (LJF), a key traditional herbal medicine, and aims to evaluate how ecological factors impact its quality. We developed a new evaluation method combining high-performance liquid chromatography (HPLC) fingerprinting technology and MaxEnt models to assess the effects of ecological factors on LJF quality. The MaxEnt model was used to predict suitable habitats for current and future scenarios, while HPLC was employed to analyze the contents of key compounds. We also used ArcGIS for spatial analysis to create a quality zoning map. The analysis identified 21 common chromatographic peaks, with significant variations in the contents of Hyperoside, Rutin, Chlorogenic acid, Cynaroside, and Isochlorogenic acid A across different habitats. Key environmental variables influencing LJF distribution were identified, including temperature, precipitation, and elevation. The current suitable habitats primarily include regions south of the Yangtze River. Under future climate scenarios, suitable areas are expected to shift, with notable expansions in southern Gansu, southeastern Tibet, and southern Liaoning. The spatial distribution maps revealed that high-quality LJF is predominantly found in central and southern Hebei, northern Henan, central Shandong, central Sichuan, southern Guangdong, and Taiwan. The study indicates that suitable growth areas can promote the accumulation of certain secondary metabolites in plants, as the accumulation of these metabolites varies. The results underscore the necessity of optimizing quality based on cultivation practices. The integration of HPLC fingerprinting technology and the MaxEnt model provides valuable insights for the conservation and cultivation of herbal resources, offering a new perspective on evaluating the impact of ecological factors on the quality of traditional Chinese medicines.

Effective strategy for distinguishing raw and vinegar Schisandrae Chinensis Fructus based on electronic eye and electronic tongue combined with chemometrics.

Schisandrae Chinensis Fructus (SCF), a traditional Chinese medicine, has been used in treating virtual injury and strain since ancient times. The Chinese Pharmacopoeia reveals that SCF includes raw (RSCF) and vinegar-processed (VSCF) decoction pieces. This study developed an effective method combining the electronic eye (e-eye), electronic tongue (e-tongue), and chemometrics to discriminate RSCF and VSCF from the perspective of chemical composition, color, and taste. First, RSCF were collected and processed into VSCF, and their color parameters, e-tongue sensory properties, high-performance liquid chromatography (HPLC) and ultra-HPLC (UPLC) characteristic fingerprints, and nominal ingredients were determined. Multivariate statistical analyses, including principal component, linear discriminant, similarity, and partial least squares discriminant analyses, were conducted. HPLC and UPLC fingerprints were established, demonstrating a > 0.900 similarity. The content determination indicated increased schisantherin A, schisantherin B, and schisandrin A contents in VSCF. The e-eye data demonstrated a > 1.5 total color difference before and after processing ΔE*ab, indicating the significantly changed sample color and appearance before and after processing. The e-tongue technology was used to quantitatively characterize the taste of RSCF and VSCF. The t-test revealed significantly reduced sourness, aftertaste-bitter, and aftertaste-astringent values of SCF after vinegar processing. Principal component and partial least squares discriminant analyses indicated that e-eye and e-tongue realize the rapid RSCF and VSCF identification. The proposed comprehensive strategy of electronic eye and electronic tongue combined with chemometrics demonstrated satisfactory results with high efficiency, accuracy, and reliability. This can be developed into a novel and accurate method for discriminating RSCF and VSCF.

Establishment of an HPLC Fingerprint and Cluster Analysis for Miao Ethnic Medicine Osbeckia opipara

: Osbeckia opipara, a traditional Miao medicine, is commonly used by the renowned national-level Chinese Traditional Medicine practitioner Zhengshi Wu for the treatment of diarrhea due to its strong antioxidative, anti-inflammatory, and antidiarrheal effects. This study aimed to establish a high-performance liquid chromatography (HPLC) fingerprint for O. opipara to provide new evidence and technical means for the scientific evaluation and effective quality control of O. opipara. The procedure involved isolation with a Nano ChromCore C18 column (250 mm × 4.6 mm, 5 μm), using a gradient elution of 0.1% formic acid in water and 0.1% formic acid in acetonitrile as the mobile phase, with a flow rate of 1.0 mL/min, a column temperature of 30°C, an injection volume of 10 μL, and detection at a wavelength of 254 nm. Under these chromatographic conditions, fingerprint analysis was conducted on 11 batches of O. opipara collected from different origins. The National Pharmacopoeia Committee developed the 'Chromatographic Fingerprint Similarity Evaluation System' (2004A version) for automated comparison, similarity computation, and analysis of chromatographic data. The results revealed 13 common peaks across the 11 batches of O. opipara samples, with a similarity to the automatically generated reference spectrum exceeding 0.9. SPSS 26.0 software was used to conduct cluster analysis on the peak areas of the 13 common peaks. The observations indicated that the reference spectrum generated from the 11 batches could serve as the standard fingerprint profile for O. opipara, providing sufficient characteristic information extraction.

Quality evaluation of WuweiZhenju tablets based on HPLC fingerprint, content determination and chemical pattern recognition analysis

AbstractObjectiveTo establish a method for the high-performance liquid chromatography (HPLC) fingerprint and content determination of Wuwei Zhenju Tablets, and combine with chemical pattern recognition analysis to provide basis for its quality evalution.MethodsBased on HPLC, taking the chromatographic peak of puerarin as the reference peak, the fingerprints of 10 batches of Wuwei Zhenju Tablets were established and the similarity evaluation was carried out to determine the common peaks. SPSS 26.0 and SIMCA 14.1 software were used to evaluate the overall quality of Zhenju Tablets by cluster analysis and principal component analysis. The contents of chlorogenic acid, puerarin, daidzin, isochlorogenic acid B, isochlorogenic acid A and aurantio-obtusin in the samples were determined by the same method.ResultsTwenty-six common peaks were identified in the fingerprints of 10 batches of Wuwei Zhenju Tablets, and the similarity evaluation was greater than 0.985. Ten chromatographic peaks were identified as neochlorogenic acid, chlorogenic acid, caffeic acid, puerarin, daidzin, isochlorogenic acid A, isochlorogenic acid B, isochlorogenic acid C, aurantio-obtusin and betaine. Combined with cluster analysis and principal component analysis, 10 batches of samples can be clustered into two categories, which can distinguish different Zhenju tablets. The model established in pattern recognition research can accurately identify Wuwei Zhenju tablets, and the prediction results are ideal. The six index components had a good linear relationship in their respective concentration ranges. The contents of chlorogenic acid, puerarin, daidzin, isochlorogenic acid B, isochlorogenic acid A and aurantio-obtusin in 10 batches of Wuwei Zhenju Tablets were 2.902–5.819, 3.693–49.680, 1.449–10.654, 2.768–7.724, 2.941–6.343 and 0.071–0.089 mg g−1, respectively.ConclusionThe HPLC fingerprint and multi-component content determination method of Wuwei Zhenju Tablets were established for the first time. The method is simple, efficient and accurate, and can effectively provide experimental basis for the quality control and evaluation of the preparation.

Spectrum-effect relationship between HPLC fingerprint and hypoglycemic of litchi leaves (Litchi chinensis Sonn) in vitro.

Litchi chinensis Sonn (Litchi) has been listed in the Chinese Pharmacopeia, and is an economically and medicinally valuable species within the family Sapindaceae. However, the material basis of its pharmacological action and the pharmacodynamic substances associated with its hypoglycemic effect are still unclear. The predominant objective of this study was to establish the fingerprint profile of litchi leaves and to evaluate the relationship between the components of the high-performance liquid chromatography (HPLC) fingerprint of litchi leaves, assess its hypoglycemic effect by measuring α-glucosidase and α-amylase inhibition, and find the spectrum-effect relationship of litchi leaves by bivariate correlation analysis, Grey relational analysis and partial least squares regression analysis. In this study, the fingerprint of litchi leaves was established by HPLC, and a total of 15 common peaks were identified that clearly calibrated eight components, with P1 being gallic acid, P2 being protocatechuic acid, P3 being catechin, P6 being epicatechin, P12 being rutin, P13 being astragalin, P14 being quercetin and P15 being kaempferol. The similarities between the fingerprints of 11 batches of litchi leaves were 0.766-0.979. Simultaneously, the results of the spectrum-effect relationship showed that the chemical constituents represented by peaks P8, P3, P12, P14, P2, P13, and P11 were relevant to the hypoglycemic effect.

VG111: A novel formulation demonstrating clinical evidence of anti-pathogenic activity and accelerated wound healing in humans and companion animals

Objectives: The increasing incidence of chronic wounds such as diabetic foot ulcers and pressure ulcers, often compounded by bacterial infections and biofilm formation, presents significant challenges in wound management. Despite advancements in wound care products and a better understanding of molecular wound repair mechanisms, the treatment of chronic ulcerating conditions remains incomplete. VG111, a novel natural product formulation, emerges as a promising therapeutic candidate addressing the need for an effective wound healing agent with antimicrobial and tissue regenerative properties. Materials and Methods: A thorough evaluation of VG111 included antimicrobial assays to determine its minimum inhibitory concentration against an array of pathogens, assessment of its biofilm disruption capabilities, investigation into its profibrogenic activity through scratch assays, and analysis of its immunomodulatory effects on macrophage-derived cytokines. Quality consistency was ensured by high-performance liquid chromatography fingerprinting, while clinical applicability was assessed through observations in canine and human wound healing cases. Statistical Analysis: The cytotoxic effects of VG111 were assessed using a Two-way ANOVA, indicating no significant cytotoxicity at the tested concentration (Column factor p<0.0001). Results: VG111 demonstrated potent antimicrobial action with effective concentrations ranging from 2.5% to 5.0% v/v, targeting resistant strains of Methicillin-resistant Staphylococcus aureus, colistin-resistant Escherichia coli, Acinetobacter baumannii, and other priority pathogens. It showed biofilm clearance, enhanced fibroblast migration, and a favorable immunomodulatory profile by reducing inflammatory cytokines in vitro. In vivo applications corroborated these findings, with significant wound healing observed in both veterinary and clinical settings, negating the need for additional antibiotics. Conclusions: The study emphasized on VG111 as a robust wound healing agent with significant antimicrobial and biofilm-disrupting properties. Its broad-spectrum efficacy against critical pathogens and ability to promote tissue regeneration mark it as a promising avenue in the management of complex chronic wounds, meriting further clinical exploration.

Chemical constituent characterization and determination of Quisqualis fructus based on UPLC-Q-TOF-MS and HPLC combined with fingerprint and chemometric analysis.

Quisqualis fructus (QF) is a traditional Chinese medicine (TCM) that it has a long history in the therapeutic field of killing parasites, eliminating accumulation, and stopping diarrhea. However, the therapeutic material basis of QF is remaining ambiguous nowadays. The geographical origin differences of QF are also usually ignored in the process of medication. In this study, the alcohol-aqueous soluble constituents in QF from different origins were systematically characterized and accurately measured by ultra-high performance liquid chromatography coupled to quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS) and high-performance liquid chromatography (HPLC) respectively. Chemometric analysis was performed for origin differentiation and screening of potential quality marker (Q-marker). Finally, A total of 106 constituents were tentatively characterized in positive and negative ion modes, including 29 fatty acids, 26 organic acids, 11 amino acids and derivatives, 10 glycosides, 9 alkaloids and derivatives, and 21 other constituents. QF from different origins were effectively distinguished and 16 constituents were selected as the potential Q-markers subsequently. Four representative components (trigonelline, adenosine, ellagic acid, and 3,3'-di-O-methylellagic acid) in QF samples were simultaneously determined. HPLC fingerprint analysis indicated that the similarity between 16 batches of QF was in the range of 0.870-0.999. The above results provide some insights for the research on the pharmacodynamic constituents, quality control, and geographical discrimination of QF.

Proximate Composition, Comparative Phytochemical Analysis, and HPLC Profiling of Various Solvent Extracts of Anthocleista djalonensis Leaves

Choosing the right solvent in herbal preparation is very important for extracting bioactive compounds from medicinal plant, as it greatly affects the yield and quality of phytochemicals obtained. This study aimed to investigate the proximate composition, phytochemical analysis and HPLC profiling of various solvents extracts of Anthocleista djalonensis leaves, a medicinal plant used in herbal remedies. The dried leaves of Anthocleista djalonensis were analyzed for their nutritional profile using standard methods. The leaves were then extracted by maceration using three different solvents: 95% analytical ethanol, hot distilled water, and local gin (78% alcohol). Various phytochemicals in the extracts were identified and quantified using standard analytical methods. Additionally, High-Performance Liquid Chromatography (HPLC) was used to compare the chemical compositions of the extracts, using a reverse-phase C18 column and UV detection at 254nm. The results of the proximate analysis revealed good nutritional value, with protein (15.90%), moisture content (9.08%), total ash (10.37%), crude fibre (4.00%), crude fat (2.40%), carbohydrate (61.53%), and an energy value of 331.33 KJ. All tested phytochemicals were found in the ethanol and local gin extracts, while the distilled water extract showed the absence of terpenoids and steroids. In the quantitative analysis, the local gin extract had the highest concentrations of alkaloids, terpenoids, flavonoids, and tannins. On the other hand, the HPLC fingerprint analysis identified seventeen (17), twelve (12), and nine (9) compounds in ethanol, local gin and water extracts respectively. Four (4) shared peaks with similar retention time were observed across all extracts, indicating the presence of common phytocompounds. In conclusion, this study shows that Anthocleista djalonensis leaves have good nutritional values, suggesting their potential as dietary phytonutrient. Local gin stands out as the best solvent among those tested for extracting beneficial compounds from Anthocleista djalonensis leaves supporting its traditional use as solvent in herbal preparation.

The effect of ozone sterilization technology on the quality of Ganmaoling Granule based on HPLC fingerprint, chemometric analyses and multi-component content determination

Ganmaoling Granule (GMLG) is one of the most effective Chinese Patent Medicines (CPMs) which is applied to relieve the uncomfortable symptoms caused by the common cold. In this study, high-performance liquid chromatography (HPLC) combined with chemometric analyses and multi-component content determination were performed to preliminary investigate the effect of ozone sterilization technology on the quality of GMLG. The methodology validation and similarity analysis results showed that the analytical method was capable of complying with the demand of HPLC fingerprints, and 80 batches of GMLGs before and after ozone sterilization in four process stages had a high degree of similarity based on the values of similarity analysis (SA) was greater than 0.98. The results of hierarchical cluster analysis (HCA) and principal component analysis (PCA) showed that the GMLGs before and after ozone sterilization were divided into two categories, and the same batch of GMLGs was clustered into the same group. Meanwhile, the contents of linarin, caffeine, chlorpheniramine maleate and acetaminophen between 24 batches of GMLGs before and after ozone sterilization had no statistically significant differences (P > 0.05), which met the requirements of “Drug Standard of the People’s Republic of China”. In conclusion, the ozone sterilization technology had no obvious influence on the quality of GMLGs in four process stages, which provided a reference for the quality control of GMLG and other Chinese-western medicine compound preparations.

Notoginseng leaf triterpenes promotes angiogenesis by activating the Nrf2 pathway and AMPK/SIRT1-mediated PGC-1/ERα axis in ischemic stroke

Notoginseng leaf triterpenes (PNGL), derived from the dried stems and leaves of P. notoginseng, is a phytoestrogen that exerts many neuroprotective effects in vivo and in vitro of ischemic stroke. However, its impact on neurological restoration specifically in relation to angiogenesis following ischemic stroke remains unexplored. The aim of this study was to assess the effects of PNGL on angiogenesis subsequent to ischemic stroke. Male Sprague-Dawley rats were utilized in this study and were subjected to middle cerebral artery occlusion/reperfusion (MCAO/R). Post-ischemia, PNGL were administered through intraperitoneal (i.p.) injection. The high-performance liquid chromatography (HPLC) fingerprinting, triphenyltetrazolium chloride (TTC) staining, immunofluorescent staining, network pharmacology and western blot analyses were assessed to determine the therapeutical effect and molecular mechanisms of PNGL on cerebral ischemia/reperfusion injury. Our findings demonstrate that PNGL effectively reduced infarct volume, enhanced cerebral blood flow, and induced angiogenesis in rats subjected to MCAO/R. Notably, PNGL also facilitated neuronal proliferation and migration in HUMECs in vitro. The proangiogenic effects of PNGL were found to be linked to the activation of the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway and the AMPK/SIRT1-mediated PGC-1/ERα axis, as well as the activation of neurological function. Our study provides evidence that PNGL hold promise as an active ingredient of inducing proangiogenic effects, potentially through the activation of the Nrf2 pathway and the AMPK/SIRT1-mediated PGC-1/ERα axis. These findings contribute to the understanding of novel mechanisms involved in the restoration of neurological function following PNGL treatment for ischemic stroke.