Chromatographic Fingerprint Research Articles

Target Cell Extraction and Spectrum–Effect Relationship Coupled with BP Neural Network Classification for Screening Potential Bioactive Components in Ginseng Extract with a Protective Effect against Myocardial Damage

Cardiovascular disease has become a common ailment that endangers human health, having garnered widespread attention due to its high prevalence, recurrence rate, and sudden death risk. Ginseng possesses functions such as invigorating vital energy, enhancing vein recovery, promoting body fluid and blood nourishment, calming the nerves, and improving cognitive function. It is widely utilized in the treatment of various heart conditions, including palpitations, chest pain, heart failure, and other ailments. Although numerous research reports have investigated the cardiovascular activity of single ginsenoside, there remains a lack of systematic research on the specific components group that predominantly contribute to cardiovascular efficacy in ginseng medicinal materials. In this research, the spectrum–effect relationship, target cell extraction, and BP neural network classification were used to establish a rapid screening system for potential active substances. The results show that red ginseng extract (RGE) can improve the decrease in cell viability and ATP content and inhibit the increase in ROS production and LDH release in OGD-induced H9c2 cells. A total of 70 ginsenosides were identified in RGE using HPLC-Q-TOF-MS/MS analysis. Chromatographic fingerprints were established for 12 batches of RGE by high-performance liquid chromatography (HPLC). A total of 36 common ingredients were found in 12 batches of RGE. The cell viability, ATP, ROS, and LDH of 12 batches RGE were tested to establish gray relationship analysis (GRA) and partial least squares discrimination analysis (PLS-DA). BP neural network classification and target cell extraction were used to narrow down the scope of Spectral efficiency analysis and screen the potential active components. According to the cell experiments, RGE can improve the cell viability and ATP content and reduce the oxidative damage. Then, seven active ingredients, namely, Ginsenoside Rg1, Rg2, Rg3, Rb1, Rd, Re, and Ro, were screened out, and their cardiovascular activity was confirmed in the OGD model. The seven ginsenosides were the main active substances of red ginseng in treating myocardial injury. This study offers a reference for quality control in red ginseng and preparations containing red ginseng for the management of cardiovascular diseases. It also provides ideas for screening active ingredients of the same type of multi-pharmacologically active traditional Chinese medicines.

Identification of Codonopsis Radix from different sources by integrating high‐performance liquid chromatography fingerprinting with chemometric approaches and Lobetyolin determination

AbstractCodonopsis Radix (CR) is often used as both a medicine and a food ingredient. However, identifying CR can be difficult due to various factors such as variety, sulfur fumigation, and storage time. Consequently, a comprehensive strategy based on fingerprinting and chemometric approaches was developed to distinguish different CRs. First, the fingerprints of CR were acquired by high‐performance liquid chromatography (HPLC). Secondly, various chemical pattern recognition methods were employed to identify CR. Additionally, 14 and 16 major compounds were identified as marker compounds to differentiate between different varieties and sulfur‐fumigated samples, respectively. Furthermore, the amount of Lobetyolin present in different samples was determined, showing that the content of Lobetyolin varied significantly across the samples. Codonopsis pilosula Nannf. var. modesta (Nannf.) L.T.Shen had the highest content of Lobetyolin, followed by Codonopsis tangshen Oliv., and Codonopsis pilosula (Franch.) Nannf. In addition, the content of Lobetyolin was highest when it was non‐sulfur‐fumigated and stored for 0 years. Conclusively, HPLC fingerprint in conjunction with chemical pattern recognition and component content determination, can be employed to differentiate different varieties and sulfur‐fumigated CR. Additionally, it is a reliable, comprehensive, simple, and rapid method for the identification of CR.

Discrimination between the Triglyceride Form and the Ethyl Ester Form of Fish Oil Using Chromatography-Mass Spectrometry.

Although the triglyceride form is the natural form of fish oil found in fish, the ethyl ester form of fish oil, which is used during processing to save costs, is also present on the market. In this study, fatty acids and lipids were determined using gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-linear ion trap mass spectrometry (LC-LIT/MS), respectively, according to developed methods. The identification of fatty acids was based on the mass spectral characteristics and equivalent chain lengths. However, the fatty acid contents of both forms of fish oils are quite similar. The application of the LC-LIT/MS method for the structural characterization of triacylglycerols (TAGs) and the mechanism of LIT/MS fragmentation are also discussed. Neutral losses of CH2=CH2 (m/z 28) and CH3CH2OH (m/z 46), which are LIT/MS characteristics of ethyl ester from fish oil, were found for the first time. The triglyceride form of fish oils was easily and accurately identified using fingerprint chromatography. In conclusion, lipid analysis combined with LC-LIT/MS showed an improved capability to distinguish between types of fish oil.

Quality control methods for fruit extracts of Kigelia africana using high performance thin layer chromatography

Kigelia africana is a tree native to Africa but also found in eastern and southern parts of India with reported anti-bacterial, anti-inflammatory, and immunomodulatory activities. Verbascoside, caffeic acid and ferulic acid are important markers for the quality control of the plant. Two different HPTLC methods were developed and validated; method − 1 for estimation of verbascoside and caffeic acid while method − 2 for estimation of caffeic acid and ferulic acid. Developed methods were applied to the methanolic fruit extract to determine the quantities of markers. Both methods were found to be linear, specific, precise, accurate, sensitive and robust. Results indicated that both methods can be used for quantitative determination of verbascoside, caffeic acid and ferulic acid in fruit extract. The developed methods may be utilised as a part of the quality control and standardisation for the raw material and extracts of Kigelia africana and can also aid to chromatographic fingerprinting of the plant.

High performance thin layer chromatography fingerprint on cyano-bonded stationary phase of selected Stachys and Betonica species and their antioxidant activity with chemometric calculations

The methanolic extracts of selected Stachys and Betonica species were analyzed by high performance thin layer chromatography (HPTLC) method using cyano bonded stationary phase and mobile phase consisted of methanol and water 45:55 (v/v) with Natural product reagent derivatization. The HPTLC method was validated (linearity, precision, accuracy, robustness, LOD, LOQ) and the validation results confirm the usefulness of the method used. The obtained images of fingerprint chromatograms presented as numerical data (using the TLC analyzer program) were next processed using Excel and SpecAlign softwares. The phytochemical similarity between analyzed plant extracts was evaluated using the similarity index (Pearson correlation coefficient) and the results were presented as hierarchical cluster analysis dendrograms and as graph for principal component analysis (PCA). The total phenolic content (TPC) by Folin-Ciocalteu method and the antioxidant activity with Ferric Ion Reducing Antioxidant Potential (FRAP) technique were determined for studied extracts using gallic acid as standards in both methods.

SIRT1/P53 in retinal pigment epithelial cells in diabetic retinopathy: a gene co-expression analysis and He-Ying-Qing-Re formula treatment.

Diabetic retinopathy (DR) is a severe diabetic complication that leads to severe visual impairment or blindness. He-Ying-Qing-Re formula (HF), a traditional Chinese medicinal concoction, has been identified as an efficient therapy for DR with retinal vascular dysfunction for decades and has been experimentally reported to ameliorate retinal conditions in diabetic mice. This study endeavors to explore the therapeutic potential of HF with key ingredients in DR and its underlying novel mechanisms. Co-expression gene modules and hub genes were calculated by weighted gene co-expression network analysis (WGCNA) based on transcriptome sequencing data from high-glucose-treated adult retinal pigment epithelial cell line-19 (ARPE-19). The chromatographic fingerprint of HF was established by ultra-performance liquid chromatography coupled with high-resolution mass spectrometry (UPLC-Q-TOF-MS). The molecular affinity of the herbal compound was measured by molecular docking. Reactive oxygen species (ROS) was measured by a DCFDA/H2DCFDA assay. Apoptosis was detected using the TUNEL Assay Kit, while ELISA, Western blot, and real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) were used for detecting the cytokine, protein, and mRNA expressions, respectively. Key compounds in HF were identified as luteolin, paeoniflorin, and nobiletin. For WGCNA, ME-salmon ("protein deacetylation") was negatively correlated with ME-purple ("oxidative impairment") in high-glucose-treated ARPE-19. Luteolin has a high affinity for SIRT1 and P53, as indicated by molecular docking. Luteolin has a hypoglycemic effect on type I diabetic mice. Moreover, HF and luteolin suppress oxidative stress production (ROS and MDA), inflammatory factor expression (IL-6, TNF-α, IL1-β, and MCP-1), and apoptosis, as shown in the in vivo and in vitro experiments. Concurrently, treatment with HF and luteolin led to an upregulation of SIRT1 and a corresponding downregulation of P53. Using HF and its active compound luteolin as therapeutic agents offers a promising approach to diabetic retinopathy treatment. It primarily suppressed protein acetylation and oxidative stress via the SIRT1/P53 pathway in retinal pigment epithelial cells.

Systematic review on fingerprinting development to determine adulteration of Chinese herbal medicines

BackgroundIt has been a current research hospots using fingerprinting technology for quality control of Chinese herbal medicines (CHMs), which provides a scientific basis for establishment of overall quality control in accordance with the characteristics of CHMs. The fingerprinting technology for CHMs is diverse, and the research field covers many disciplines, such as analytical chemistry, pharmacology, pharmaceutics, biochemistry, and molecular biology. PurposeTo effectively understand the key areas and future directions of research regarding the fingerprint and adulteration of CHMs. Methods/resultsthis paper analyzed 879 articles in this field in the Web of Science Core Collection from 2000 to 2023 with CiteSpace and VOSviewer, and systematically assessed the research process, hotspots, topic distribution among disciplines, etc. The most prominent contributors of fingerprint and adulteration of CHMs research are mainly from China, India, the United States, England, and Brazil. The knowledge domains of fingerprint and adulteration of CHMs research focus mainly on the topics of molecular authentication, DNA barcoding, HPLC, near-infrared spectroscopy, manage data, chemometrics, and electrochemical fingerprinting. Most countries have recognized the pharmaceutical potential of natural products, and have paid more attention to the fingerprint and adulteration of CHMs in the past decade. Future the research tends to focus more on molecular identification and authentication, and electrochemical and chromatographic fingerprinting in controlling the adulteration of CHMs. ConclusionThis research provides a valuable reference for scholars in related fields to analyze existing research results, understand the development trend, and explore new research directions.

Pharmacognostical studies of Eclipta prostrata (Linn.) grown in Sri Lanka

Eclipta prostrata Linn. is an annual herbaceous plant used in traditional medicine, commonly known as “Trailing Eclipta” or False Daisy in English, ‘Keekirindiya’ in Sinhala and ‘Kayyantakara’ in Tamil. The aim of this study was to conduct a detailed pharmacognostical evaluation of E. prostrata found in Sri Lanka. Anatomical, physicochemical, phytochemical studies, and quantification of phyto-constituents were performed as per WHO guidelines. Whole plant was sequentially extracted into solvents with different polarities. Phytochemical screening and Thin Layer Chromatography (TLC) fingerprinting were carried out. Anatomical study and powder microscopy revealed useful diagnostic features. Physicochemical parameters such as moisture content, ash values (total, acid insoluble, water soluble) and extractable matter in water and ethanol were evaluated. Phytochemical screening and TLC fingerprinting revealed the presence of different types of phyto-constituents. Alkaloid, tannin, saponin, total flavonoid and total polyphenol contents were quantified. In conclusion, pharmacognostical study aids in establishing the standardization parameters of E.prostrata found in Sri Lanka.

Quality Evaluation and Identification of Phyllanthi fructus (Yuganzi) Based on the Spectrum-Effect Relationship

To establish a quality evaluation and identification method for Phyllanthi fructus (Yuganzi), the spectrum-effect relationship was explored. A high-performance liquid chromatography (HPLC) fingerprint was established using ultraviolet spectrophotometry, and the in vitro antioxidant activity was determined using a total antioxidant capacity assay kit. Similarity analysis, hierarchical cluster analysis (HCA), and partial least squares regression (PLSR) were performed to establish the spectrum-effect relationships. Thirteen batches of Yuganzi were collected for testing. The results revealed that the optimal chromatographic conditions for the HPLC fingerprint were as follows: the mobile phase consisted of 0.1% phosphoric acid solution (A) and acetonitrile (B), the detection wavelength was 214 nm, the column temperature was 30 °C, and the flow rate was 0.8 mL/min. Among the batches of samples, the similarity values of 10 samples (S1–S10) from Yunnan were larger than 0.995; the similarity values of 3 samples (S11–S13) from India, Gaoligong Mountain, and Fujian were less than or equal to 0.986. Furthermore, nineteen characteristic peaks of Yuganzi were calibrated using fingerprint evaluation software. The study on the spectrum-effect relationship further revealed that compounds corresponding to peaks 5 and 8 were potentially key ingredients for the quality evaluation and identification of Yuganzi, closely related to the stable antioxidant activities of Yuganzi. The spectrum-effect relationship is an agile and efficient approach that can ensure the intra-assay stability of Yuganzi from same region and identify Yuganzi from different regions. Compounds with antioxidant activity can be identified as quality markers for Yuganzi.

Establishment of multivariate digital high performance liquid chromatography fingerprints of fruit juice essence and its flavor quality evaluation

Quality control problem of essence and spices has always been the focus of food chemists in recent years. In this study, a multi-variate digital fingerprint profile of fruit juice essence sample was established using the combination of HPLC-DAD spectra and UPLC-MS spectra, which were collected to obtain the information about organic acids, polyphenols, sugars and sugar alcohols of the samples. The stability, reliability and repeatability of the combined fingerprinting method were verified in the work. A total of 36 markers in the established fingerprint were common (20 compounds were confirmed) belonging to different classes: organic acids, polyphenols, sugars, sugar alcohols, and other miscellaneous compounds. Those chromatographic peaks that significantly affect the quality of apple juice flavor were screened from principal component analysis (PCA). Moreover, PCA and cluster analysis of the proposed combined fingerprint gave the better evaluation of the sample quality stability and distinguish sample groups than the fingerprint from single data, which demonstrated its great significance to the quality control of fruit juice samples.

Spectrum-effect relationship between ultra-performance liquid chromatography fingerprints and melanogenic effect of Vernonia anthelmintica effective part.

The fingerprint of Vernonia anthelmintica effective part (VAEP) from 15 different producing areas was established, followed by cluster analysis and principal component analysis. The relationship between the fingerprint and the melanogenesis-promoting activity of VAEP was then analyzed using the grey correlation degree and the orthogonal partial least square method. The characteristic peaks reflecting the pharmacodynamic effect of VAEP were identified as vernodalin, 3,5-O-dicaffeoyl quinic acid (3,5-diCQA), and butin. Based on the distribution characteristics of these components in plants from different habitats and the verification of results from the spectrum-effect relationship, vernodalin and 3,5-diCQA can be used as characteristic components for quality control and pharmacodynamic assessment of V. anthelmintica products. This research establishes a theoretical foundation for planting areas and provides a scientific evaluation of the melanogenesis-promoting effect of V. anthelmintica.

Discovery of Potential Anti-Microbial Molecules and Spectrum Correlation Effect of Ardisia crenata Sims via High-Performance Liquid Chromatography Fingerprints and Molecular Docking.

Ardisia crenata Sims, an important ethnic medicine, is recorded in the Chinese Pharmacopoeia for treating laryngeal diseases and upper respiratory tract infections. This study aimed to evaluate the antimicrobial effect of extracts and potential antimicrobial compounds of A. crenata Sims. It was found that the roots of A. crenata Sims have a potential inhibitory effect on Candida albicans and Aspergillus flavus, with MICs of 1.56 mg/mL and 0.39 mg/mL, and the leaves of A. crenata Sims have a potential inhibitory effect on Pseudomonas aeruginosa and Staphylococcus aureus, with MICs of 3.12 mg/mL and 6.77 mg/mL, respectively. Meanwhile, five compounds including one catechin and four bergenins were obtained from roots. These components were identified on the fingerprint spectrum, representing chromatographic peaks 16, 21, 22, 23, and 25, respectively. Among these, 11-β-d-glucopyranosyl-bergenin and (-)-gallocatechin showed potential inhibition for Staphylococcus aureus and Pseudomonas aeruginosa with MIC of 0.26 and 0.33 mg/mL, respectively. The roots, stems, and leaves of A. crenata Sims are very similar in chemical composition, with large differences in content. Principal component analysis (PCA) and Hierarchical cluster analysis (HCA) showed that 16 batches of A. crenata Sims could be divided into four main production areas: Guizhou, Jiangsu, Guangxi, and Jiangxi. Furthermore, molecular docking results showed that 11-β-d-glucopyranosyl-bergenin had a better affinity for Casein lytic proteinase P (ClpP), and (-)-gallocatechin possessed a strong affinity for LasA hydrolysis protease and LasB elastase. These findings suggest catechin and bergenins from A. crenata Sims can be used as antimicrobial activity molecules.

In-vitro, in-vivo and in-silico anti-inflammatory evaluation of hedychenone and its semisynthetic derivative

Hedychium spicatum is a perennial shrub that is commonly found in the Himalayan region. Thin layer chromatography fingerprint screening of various Hedychium species revealed that only the species Hedychium spicatum possesses hedychenone. Purification and isolation of hedychenone was carried out from methanolic extract of Hedychium spicatum rhizomes powder using column chromatography. After crystallization of enriched fraction, 2.19% of hedychenone crystalline powder was yielded. The physiochemical, pharmacokinetic, and pharmacodynamic evaluation of the active pharmaceutical components was conducted using artificial learning and machine learning (AL/ML) approaches. The results indicate that both medications adhere to Lipinski’s rule of five. The docking study indicated that compound 2 (MBE-10.4 kcal/mol) exhibited a more sub-stantial interaction with the cannabinoid receptor 2 compared to hedychenone (MBE -9.85 kcal/mol). A study was done to assess the physical and chemical stability of hedychenone under biological pH conditions. Furthermore, the administration of both medication concentrations, 100 mg/kg and 200 mg/ kg, effectively inhibits the development of acute paw edema caused by ovalbumin. A preliminary investigation of hedychenone and its synthetic derivative indicated that these compounds have the potential to be utilized as active medicinal ingredients in herbal pharmaceutical preparations.

Application of polyphenolic compound-based HPLC fingerprint in Chinese golden camellias (Camellia sect. Chrysantha)

Chinese golden camellias (Camellia sect. Chrysantha) are traditional substitute tea with medicinal and food properties. The development of fingerprinting techniques specific to Chinese golden camellias may provide a new means of reliably assessing the quality of tea and traditional medicine. Herein, 32 total specimens were collected in Southern China and used to establish a polyphenolic compound-based high-performance liquid chromatography (HPLC) signature profile. The established chromatographic fingerprints were subjected to similarity, hierarchical clustering analysis (HCA), together with principal component analysis (PCA) evaluations, and the uncertainty and reliability for such HPLC fingerprints were also evaluated. Similarity values for C. sect. Chrysantha samples ranged from 0.721 to 0.969 relative to the reference chromatographic fingerprint (RFP). HCA and PCA approaches were performed to classify 13 specimens of C. sect. Chrysantha from different species and regions into four categories that respectively contained 7, 1, 2, and 3 specimens. In this HPLC fingerprint, peak 11 was authenticated as rutin, while peaks 2, 4, 6, 7, 9, 12 and 13 were tentatively characterized via liquid chromatography-high-resolution mass spectrometry (LC-HRMS) as corresponding with other polyphenolic agents. Credibility analyses indicated that the macroscopic qualitative and quantitative reliability values for C. fascicularis RFP were greater than 0.9500. Taken together, statistical analyses thus confirmed that this HPLC fingerprinting strategy can be reliably used to classify and assess the quality of C. sect. Chrysantha specimens. And the qualitative and quantitative reliability analysis and systematic quantitative fingerprint methods are promising tools for future efforts to ensure the quality of C. fascicularis.

Screening and Identification Quality Markers for Antioxidant Activity of Tetrastigma hemsleyanum from Different Origins Using Spectrum-Effect Correlation Analysis

Background:: Tetrastigma hemsleyanum (THL) is a traditional medicinal plant with various biological activities, it has been validated that several THL extracts have strong pharmacological activities. Until now, there is no complete and systematic germplasm identification and quality evaluation standard. Objective:: Establishing fingerprint spectra of THL from different sources, evaluating their antioxidant activity and identifying quality markers. Methods:: In this paper, establishing fingerprints and identifying quality markers through UPLC-QTOF- MS/MS. The antioxidant capacity of forty-one batches of THL from multiple regions was determined by 1,1-Diphenyl-2-picrylhydrazyl radical (DPPH), 2,2'-Azinobis-(3-ethylbenzthiazoline-6- sulphonate) (ABTS), and Ferric reducing antioxidant power (FRAP) assays. Results:: The results indicated that THL methanol extracts showed good antioxidative activity and eight THL samples, including THL aerial parts, showed high antioxidative activity. Besides, Ultra Performance Liquid Chromatography (UPLC) fingerprints of THL from different places were established and chromatographic peaks probably correlated with antioxidant capacity were screened by principal component analysis (PCA) and partial least square regression (PLS) analysis. Conclusion:: The chemical constituents of these peaks were identified as possible quality markers for THL quality evaluation. In this study, twelve effective substances were preliminarily determined, such as isovitexin, L-epicatechin, procyanidin B, sitosterol, etc.

Pharmacognostic study and preliminary phytochemical investigation of Andrographis paniculata (Burm.f.) Wall. ex Nees

Andrographis paniculata (family –Acanthaceae) is an annual herbaceous plant with branches that grows up to a height of 50-150 cm in moist, shaded areas. Its stem is abruptly quadrangular, heavily branched, and has a delicate texture that is readily broken. It is found in Southeast Asia, tropical and subtropical Asia, and a few other nations like Vietnam, Cambodia, and the Caribbean islands, Indonesia, Laos, Malaysia, Myanmar, Sri Lanka and Thailand etc. It is widely used in different traditional medicinal systems or countries for medicinal purposes by the traditional practitioners, tribes or community as a folklore remedies. The present communication provides a detailed account of the pharmacognostic study carried out on various parts of the Andrographis paniculata. The study includes macroscopy, microscopy, powder microscopic studies, physicochemical tests, preliminary phytochemical screening, development of HPTLC (High Performance Thin Layer Chromatography) fingerprints profile and heavy metal tests. Physicochemical parameters were performed and found average values of three parts (root, stem and leaf) such as root loss on drying at 1050C 4.93% w/w, total ash value 10.16% w/w, acid insoluble ash value 0.21% w/w, alcohol soluble extractive value 13.98% w/w and water soluble extractive value 25.77%w/w. Stem loss on drying at 1050C 6.23% w/w, total ash value 6.88% w/w, acid insoluble ash value 0.25% w/w, alcohol soluble extractive value 18.46% w/w and water soluble extractive value 21.94%w/w and leaf loss on drying at 1050C was found 6.29% w/w, total ash value 7.30% w/w, acid insoluble ash value 0.13% w/w, alcohol soluble extractive value 21.93% w/w and water soluble extractive value 38.68%w/w. HPTLC (High Performance Thin Layer Chromatography) fingerprints profile of methanolic extract was done by using mobile phase toluene: ethyl acetate (7:3). Andrographolide and Ferullic acid standard markers were applied, major spots Rf values with colour were recorded before derivatization at 254nm. Rf values are 0.25 black, leaf, stem and root with Andrographolide standard marker, and 0.20 light black color of stem, and root with Ferullic acid standard marker. Andrographolide is higher present in leaf which was range from 1.51-162 than the stem range1.46-153 and root range 1.45-1.150, while Ferulic acid was higher present in stem range from 0.91-0.96 than root 0.76-0.82 but absent in leaf. Heavy metals i.e Pb, Cd, As, & Hg were tested and found under WHO limits Established parameters can be used as standards for quality control and identification of the plant in herbal compound formulations and also preparation of a monograph of the plant.

HPLC Fingerprint Analysis of Cibotii rhizoma from Different Regions and Identification of Common Peaks by LC-MS.

To establish the fingerprint of Cibotii rhizoma using high-performance liquid chromatography (HPLC) and evaluate the quality of Cibotii rhizoma from different regions using chemometrics to identify the potential quality markers, thirteen batches of Cibotii rhizoma samples were analyzed. the similarity evaluation system of TCM chromatographic fingerprint similarity evaluation was used to confirm common peaks. The SPSS 27 software was used for hierarchical cluster analysis (HCA), and SIMCA 14.1 software was used for principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). Moreover, a batch of Cibotii rhizoma was selected for LC-MS analysis and speculated on 15 common components. HPLC fingerprint were established, 15 common peaks were matched, two chromatographic peaks were identified using standard substances (protocatechuic acid and protocatechuic aldehyde), and 13 common components were inferred through liquid chromatograph-mass spectrometer (LC-MS). The 13 batches of the samples showed good similarities (>0.910). The results of HCA, PCA and OPLS-DA showed that 13 batches of samples were divided into three groups, and different markers were selected. The method is simple, rapid and reproducible, and can provide a reference for the overall quality evaluation of Cibotii rhizoma.

Spectrum-effect relationship study between ultra-high-performance liquid chromatography fingerprints and anti-hepatoma effect in vitro of Cnidii Fructus.

Cnidii Fructus, derived from the dried ripe fruit of Cnidium monnieri (L.) Cuss, has the effect of warming kidneys and invigorating Yang. This study established the spectrum-effect relationships between ultra-high-performance liquid chromatography (UHPLC) fingerprints and the antitumor activities of Cnidii Fructus on human hepatocellular carcinoma (HepG2) cells. In UHPLC fingerprints, 19 common peaks were obtained, and 17 batches of herbs had similarity >0.948. In Cell Counting Kit-8 (CCK-8) test, 17 batches of Cnidii Fructus extract significantly inhibited the proliferation of HepG2 cells to different degrees, showing different half-maximal inhibitory concentration (IC50 ) values. Furthermore, gray correlation analysis, Pearson's analysis, and orthogonal partial least squares discriminant analysis were performed to screen out eight components. The analysis of mass spectrum data and a comparison with standards revealed that the eight components were methoxsalen, isopimpinellin, osthenol, imperatorin, osthole, ricinoleic acid, linoleic acid, and oleic acid. The verification experiments by testing single compounds indicated that these eight compounds were the major anti-hepatoma compounds in Cnidii Fructus. This work provides a model combining UHPLC fingerprints and antitumor activities to study the spectrum-effect relationships of Cnidii Fructus, which can be used to determine the principal components responsible for the bioactivity.

Quality assessment and Q-markers discovery in Citri Sarcodactylis Fructus by integrating serum pharmacochemistry and network pharmacology.

Citri Sarcodactylis Fructus (CSF), a common fruit and traditional Chinese medicine (TCM), has been hindered in its further development and research owing to the lack of comprehensive and specific quality evaluation standards. This study aimed to establish clear TCM quality standards related to the therapeutic mechanisms of CSF and to provide a basis for subsequent research and development. Ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap high-resolution mass spectrometry (UPLC-Q-orbitrap HRMS) technology was used to comprehensively identify CSF components and explore their absorbance levels in rat serum. Network pharmacology research methods were employed to investigate the potential mechanisms of action of the identified components in the treatment of major clinical diseases. Subsequently, a combination of HPLC chromatographic fingerprinting for qualitative analysis and multi-index content determination was used to evaluate the detectability of the identified quality markers (Q-markers). Twenty-six prototype components were tentatively characterized in rat serum. Network pharmacology analysis showed six effective components, namely 7-hydroxycoumarin, isoscopoletin, diosmin, hesperidin, 5,7-dimethoxycoumarin, and bergapten, which played important roles in the treatment of chronic gastritis, functional dyspepsia, peptic ulcer, and depression and were preliminarily identified as Q-markers. The results of content determination in 15 batches of CSF indicated significant differences in the content of medicinal materials from different origins. However, compared with the preliminarily determined Q-markers, all six components could be measured and were determined as Q-markers of CSF. The chemical Q-markers obtained in this study could be used for effective quality control of CSF.

Study on the Fingerprints of Acanthopanax trifoliatus Fractions with Varying Polarities and the Relationship Between Anti-inflammatory and Antioxidant Activity Using Multivariate Analysis

Objectives: To evaluate the Spectrum-Effect relationships between the High-Performance Liquid Chromatography (HPLC) Fingerprints of fractions with different polarities and the anti-inflammatory and antioxidant effects of A. trifoliatus and to identify its pharmacodynamic material basis. Materials and Methods: Chemical Fingerprints of A. trifoliatus fractions with different polarities (all fractions and ethyl acetate, n-butanol and water fractions) were obtained by HPLC, and Principal Component Analysis (PCA) was performed. The foot swelling and prostaglandin E2 (PGE2) content in mice induced by carrageenan were used as anti-inflammatory markers, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2’-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) methods were used as antioxidant indicators. Moreover, the Spectrum-Effect relationship between common peaks and efficacy value was conducted by Grey Relational Analysis (GRA) and Partial Least Squares analysis (PLS). Results: PCA identified three principal components, and all fractions of A. trifoliatus had good anti-inflammatory and antioxidant effects. Based on GRA and PLS analysis, peaks 1 and 2 (neochlorogenic acid), 4 and 5 (chlorogenic acid), 8, 9, 10, 11 (isochlorogenic acid B), and 12 were involved in anti-inflammatory efficacy, and peaks 1 and 2 (neochlorogenic acid), 3, 4, and 5 (chlorogenic acid), and 6, 7, 9, 10, and 13 (isochlorogenic acid A) were involved in antioxidant efficacy. Conclusion: This study on the Spectrum-Effect relationship of different polar fractions of A. trifoliatus provides data to support the discovery of the pharmacodynamic material basis of the anti-inflammatory and antioxidant properties of A. trifoliatus.