High-performance Liquid Chromatography Fingerprint Research Articles

Targeted and Non-Targeted HPLC Analysis of Coffee-Based Products as Effective Tools for Evaluating the Coffee Authenticity.

Coffee is a very popular beverage worldwide. However, its composition and characteristics are affected by a number of factors, such as geographical and botanical origin, harvesting and roasting conditions, and brewing method used. As coffee consumption rises, the demands on its high quality and authenticity naturally grows as well. Unfortunately, at the same time, various tricks of coffee adulteration occur more frequently, with the intention of quick economic profit. Many analytical methods have already been developed to verify the coffee authenticity, in which the high-performance liquid chromatography (HPLC) plays a crucial role, especially thanks to its high selectivity and sensitivity. Thus, this review summarizes the results of targeted and non-targeted HPLC analysis of coffee-based products over the last 10 years as an effective tool for determining coffee composition, which can help to reveal potential forgeries and non-compliance with good manufacturing practice, and subsequently protects consumers from buying overpriced low-quality product. The advantages and drawbacks of the targeted analysis are specified and contrasted with those of the non-targeted HPLC fingerprints, which simply consider the chemical profile of the sample, regardless of the determination of individual compounds present.

HPLC Fingerprint Analysis of Rana chensinensis Eggs from Different Habitats and Their Antitussive Effect.

In this paper, a novel fingerprint method was established for the quality control of Rana chensinensis eggs (RE) by high-performance liquid chromatography (HPLC). Cluster analysis and principal component analysis were performed. Besides, the antitussive effect of RE was explored. The analysis was achieved on a Kromasil 100-5C18 (4.6 mm × 250 mm, 5 μm) column by gradient elution using methanol-0.1% phosphoric acid solution as the mobile phase. The influence of RE on cough latent periods and cough times of mice was investigated via an ammonia cough-inducing experiment. The validated HPLC method was precise, reproducible, and stable. The HPLC fingerprints of 10 batches of RE samples displayed 31 well-resolved common peaks in the chromatogram. Three of these peaks were identified and assigned to 1-methyl hydantoin, estradiol, and 4-cholestene-3-one. The similarities of 10 batches of samples were more than 0.95. RE from different origins could be classified into three groups via SPSS 23.0 software, suggesting RE samples from various provinces (Jilin, Liaoning, and Heilongjiang) can be well distinguished via the established method. High dose and middle dose of the RE group can significantly prolong the cough latent periods of mice (P < 0.05 or P < 0.01) and inhibit the cough times of mice (P < 0.01), indicating RE had a good antitussive effect. HPLC fingerprint combined with multicomponent determination can be an efficient and useful method for monitoring the quality of RE. This study also provided a more comprehensive strategy for the quality evaluation of RE.

Quality control strategies of medicine food homology materials based on fingerprint profiling and chemometrics: Citri Reticulata Pericarpium as an example

The study aimed to provide a reliable and feasible strategy for the comprehensive quality control of medicine food homology materials (MFHM). The high performance liquid chromatography (HPLC) fingerprints and Fourier transform mid-infrared (FT-MIR) quantized fingerprints were successfully developed to comprehensively evaluate overall quality of Citri Reticulata Pericarpium (CRP) by applying comprehensive quantified fingerprint method (CQFM). All samples were well distinguished and divided into 5 grades. In addition, through principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA), the identification ability of HPLC fingerprints and FT-MIR fingerprints on CRP with different storage years was discussed. The results showed that HPLC fingerprints combined with PCA had good discrimination ability, and the PLS-DA model established by the preprocessed FT-MIR fingerprint data could accurately distinguish and predict the storage period of CRP. Finally, based on 1, 1-diphenyl-2-picrylhydrazyl radical (DPPH•) scavenging assay, combined with bivariate correlation analysis, the fingerprint-activity relationship of offline antioxidant activity of CRP samples with the fingerprints peak were studied. In general, the comprehensive strategies provide a reliable and scientific reference scheme for the quality control of MFHM in the future.

In-vitro Antibacterial Activity of Allium sativum on Common Food Poisoning Bacteria

Background: The use of herbal medicines for food poisoning in inadequate resource countries has greatly increased with the limited establishment of antibacterial activity. This study aimed at establishing the antibacterial activity of the aqueous Allium sativum extract against Staphylococcus aureus, Escherichia coli and Salmonella typhimurium.&#x0D; Methods: Allium sativum L. (Amaryllidaceae) was purchased from a farmer in Kasese District and was authenticated. The material was then extracted using distilled water, filtered and lyophilized. Evaluation of the phytochemicals in the extract was done using standard procedures. High-Performance Liquid Chromatography (HPLC) fingerprint of the extract was conducted to monitor the reproducibility of the extract. The antibacterial activity of the aqueous garlic extract against the organisms above was evaluated and expressed as Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC).&#x0D; Results: Phytochemical screening of the garlic extract showed the presence of amino acids, alkaloids, glycosides and saponins, flavonoids, steroids and terpenoids. HPLC fingerprints showed 39 and 98 peaks at 220nm and 254nm wavelengths respectively and the presence of various phytochemical compounds in A. sativum. The extract showed antibacterial activity against S. aureus, E. coli and S. typhi.&#x0D; Conclusion: The aqueous garlic extract exhibited antibacterial activity against tested common food poisoning bacteria, with a high potency against gram-positive bacteria.

Application of HPLC Fingerprint Combined with Chemical Pattern Recognition and Multi-Component Determination in Quality Evaluation of Echinacea purpurea (L.) Moench.

Echinacea purpurea (EP) is a common medicinal material for extracting anti-RSV components. However, up to now, there has been no effective and simple method to comprehensively reflect the quality of EP. In our current study, the quality of Echinacea purpurea (L.) Moench samples from six different cultivation locations in China was evaluated by establishing a high-performance liquid chromatography (HPLC) fingerprint, combining chemical pattern recognition and multi-component determination. In this study, the chemical fingerprints of 15 common peaks were obtained using the similarity evaluation system of the chromatographic fingerprints of traditional Chinese medicine (2012A Edition). Among the 15 components, three phenolic acids (caftaric acid, chlorogenic acid and cichoric acid) were identified and determined. The similarity of fingerprints of 16 batches of Echinacea purpurea (L.) Moench samples ranged from 0.905 to 0.998. The similarity between fingerprints of five batches of commercially available Echinacea pupurea (L.) Moench and the standard fingerprint ”R” ranged from 0.980 to 0.997, which proved the successful establishment of the fingerprint. PCA and HCA were performed with the relative peak areas of 15 common peaks (peak 3 as the reference peak) as variables. Anhui and Shaanxi can be successfully distinguished from the other four cultivation areas. In addition, the index components of caftaric acid, chlorogenic acid and cichoric acid were in the range of 1.77–8.60 mg/g, 0.02–0.20 mg/g and 2.27–15.87 mg/g. The results of multi-component index content determination show that the contents of the Shandong cultivation area were higher, followed by Gansu, Henan and Hebei, and the lowest were Anhui and Shaanxi. The results are consistent with PCA and HCA, which proved that the quality of Echinacea purpurea (L.) Moench from different origins was different. HPLC fingerprint combined with chemical pattern recognition and multi-component content determination was a reliable, comprehensive and prospective method for evaluating the quality of Echinacea purpurea (L.) Moench. This method provides a scientific basis for the quality control and evaluation of Echinacea purpurea (L.) Moench.

Chromatographic fingerprint combined with quantitative analysis of multi-components by single-marker for quality control of total lignans from Fructus arctii by high-performance liquid chromatography.

The total lignans from Fructus arctii (TLFA) is a mixture of a series of lignans isolated from dried ripe fruit of Arctium lappa L. We previously reported on the pharmacological activity of TLFA that is related to diabetes. An accurate and practical TLFA quantitative analysis method for utilising it needs to be established. This study aimed to develop an effective quantitative analysis method for assessing the TLFA quality. A total of 11 marker components were confirmed by analysing the high-performance liquid chromatography fingerprints of 24 batches of TLFA samples. The samples were prepared from TLFA and structurally identified as lappaol H, lappaol C, arctiin, arctignan D, arctignan E, matairesinol, arctignan G, isolappaol A, lappaol A, arctigenin, and lappaol F. In the quantitative analysis of multi-components by the single-marker (QAMS) method and with arctiin as an internal reference substance, the content of these lignans in TLFA was simultaneously determined according to their relative correction factors with arctiin. There was no significant difference between results measured by the QAMS and traditional external standard methods. Hierarchical cluster and principal component analyses were performed to evaluate 24 TLFA batches based on the contents of 10 marker components. The results revealed that QAMS method combined with chemometric analyses could accurately measure and clearly distinguish the different quality samples of TLFA. The QAMS method is a reliable and promising quality control method for TLFA. It can provide a reference for promoting quality control of complex multi-component systems, especially for traditional Chinese medicine.

Cold-hot nature identification of Chinese herbal medicines based on the similarity of HPLC fingerprints.

The nature theory of Chinese herbal medicines (CHMs) is the core theory of traditional Chinese medicine (TCM). Cold-hot nature is an important part of CHM nature. It is found that the material basis of cold-hot nature is CHM ingredients. To test the scientific hypothesis that “CHMs with similar cold-hot nature should have similar material basis,” we explored an intelligent method for cold-hot nature identification of CHMs based on the feature similarity of CHM ingredients in this work. Sixty one CHMs were selected for cold-hot nature identification. High performance liquid chromatography (HPLC) was used to separate the chemical ingredients of CHMs and extract the feature information of CHM ingredients. A distance metric learning algorithm was then learned to measure the similarity of HPLC fingerprints. With the learned distance metric, cold-hot nature identification scheme (CHNIS) was proposed to build an identification model to evaluate the cold-hot nature of CHMs. A number of experiments were designed to verify the effectiveness and feasibility of the proposed CHNIS model. The total identification accuracy rate of 61 CHMs is 80.3%. The performance of the proposed CHNIS algorithm outperformed that of the compared classical algorithms. The experimental results confirmed our inference that CHMs with similar cold-hot nature had similar composition of substances. The CHNIS model was proved to be effective and feasible.

Comprehensive Quality Evaluation of the Root Bark of Morus alba L. Based on High-Performance Liquid Chromatography Fingerprinting and Chemometric Analyses.

The quality of the root bark of Morus alba L. (SBP) herbs currently circulating in the market is variable. In order to ensure clinical effectiveness, a high-performance liquid chromatography (HPLC) fingerprinting method combined with chemical pattern recognition should be established to control the quality of SBP herbs. The differences of 23 batches of SBP were analyzed by exploratory cluster analysis based on shared fingerprint peak data, and the results indicated that the processing method to remove the cork layer from SBP materials is an important influencing factor on SBP quality. Principal component analysis indicated that SBP samples with the cork layer removed can be clearly distinguished from samples without cork layer removal. The potential chemical markers (kuwanon G, morusin and oxyresveratrol) were screened by partial least squares discriminant analysis. Finally, the contents of the main components were determined, indicating that the processing method of SBP materials can affect content of bioactive ingredients and that cork layer removal leads to a more uniform chemometric profile. The HPLC-based chemometrics approach described here will support the development of quality standards in SBP products.

Qualitative and quantitative analysis on crocins in fruits of Gardenia species

The aim of this study was to compare crocins in the fruit of Gardenia jasminoides and Gardenia jasminoides var. radicans. Acchrom XCharge C_(18) column(4.6 mm×250 mm, 5 μm) was used for separation, with mobile phase of acetonitrile and 0.1% formic acid for gradient elution. The detection wavelength was set at 440 nm with a flow rate of 1.0 mL·min~(-1), and the column temperature was 30 ℃. The high performance liquid chromatography(HPLC) fingerprint of crocin in Gardenia species was established by testing 20 batches of G. jasminoides and 8 batches of G. jasminoides var. radicans samples from different sources, and UHPLC-ESI-Orbitrap-MS/MS technology and reference substances were used to predict and identify the common peaks. The results showed that 20 common chromatographic peaks from the samples were selected and the structures of 16 common peaks were predicted by mass spectrum. Four common peaks(crocin Ⅰ, Ⅱ, Ⅲ, and Ⅳ) were identified by the comparison with reference substances. The content of crocin Ⅰ, Ⅱ, Ⅲ, and Ⅳ was determined simultaneously under the same chromatographic condition, and both the system suitability and the methodological investigation results met the requirements of content determination. The relative similarity of HPLC fingerprint of 28 samples to the reference fingerprint was above 0.98. The results of cluster analysis(CA) showed that G. jasminoides and G. jasminoides var. radicans were separately grouped into one group. In the 20 batches of G. jasminoides, the content of crocin Ⅰ, Ⅱ, Ⅳ, and Ⅲ was between 3.58-9.58, 0.230-1.452, 0.014 5-0.135, and 0.301-1.12 mg·g~(-1), respectively, and the total content was between 4.12-12.25 mg·g~(-1). In the 8 batches of G. jasminoides var. radicans, the content of crocin Ⅰ, Ⅱ, Ⅳ, and Ⅲ was between 5.84-11.48, 0.308-0.898, 0.010 6-0.025 5, and 0.675-1.34 mg·g~(-1), respectively, and the total content was between 6.97-13.72 mg·g~(-1). The existing results showed that there is a certain similarity between G. jasminoides and G. jasminoides var. radicans in the composition of crocin, which needs further proved by more batches of samples. The method established in this paper provides references for the quality control of G. jasminoides, G. jasminoides var. radicans, and related products.

Chemical pattern recognition of Atractylodes chinensis from different producing areas and establishment of quantitative analysis of multi-components by single marker (QAMS) method for four components

This study established the fingerprint and combined it with chemical pattern recognition to evaluate the quality of Atractylodes chinensis samples from different producing areas and then employed the quantitative analysis of multi-components by single marker(QAMS) method to verify the feasibility and applicability of the established method in the quality evaluation of A. chinensis. The fingerprints of A. chinensis samples were constructed via high performance liquid chromatography(HPLC) to evaluate the inter-batch consistency. With the quality control component atractylodin as the internal reference, the relative correction factors(RCFs) were established for atractylenolide Ⅰ, atractylenolide Ⅲ, and β-eudesmol and the content of the four components was calculated. The external standard method was used to verify the accuracy of QAMS method. The quality of A. chinensis was further evaluated by similarity analysis, clustering analysis, and principal component analysis. The fingerprints of 13 batches of samples were calibrated with 21 common peaks, and 4 common peaks were identified with the similarities all above 0.9. The RCFs established with atractylodin as the internal reference represented good reproducibility under different experimental conditions. Specifically, the RCFs of atractylenolide Ⅰ, atractylenolide Ⅲ, and β-eudesmol in A. chinensis were 2.091, 4.253, and 6.010, respectively. QAMS and ESM showed no significant difference in the results, indicating that the QAMS method established in this study was stable and reliable. Thus, HPLC fingerprint combined with QAMS can be used for the quality evaluation of A. chinensis, providing a basis for comprehensive and rapid quality evaluation of A. chinensis.

Tong-Qiao-Huo-Xue decoction activates PI3K/Akt/mTOR pathway to reduce BMECs autophagy after cerebral ischemia/reperfusion injury

Ethnopharmacological relevanceTong-Qiao-Huo-Xue Decoction (TQHXD) is a traditional classic Chinese Medicinal Formula (CMF) used for clinical treatment of ischemic stroke. TQHXD leads to improvement in the symptoms of the acute period of cerebral infarction and recovery period after stroke. Our previous studies also showed that TQHXD produced a significant protective effect on the brain after cerebral ischemia-reperfusion (I/R) injury. It is reported that autophagy is closely related to ischemic brain injury; however, the functional contribution of TQHXD to brain microvascular endothelial cell (BMEC) autophagy and its underlying mechanism remains unclear. Aim of the studyThe purpose of this study was to investigate the effects and mechanism of TQHXD in inhibiting cerebral ischemia-induced endothelial autophagy. Materials and methodsThe high-performance liquid chromatography (HPLC) fingerprint of the chemical constituents from TQHXD was established for the quality control, and the Longa method was used to evaluate the efficacy of TQHXD in rats with middle cerebral artery occlusion (MCAO). The expression of LC3 was determined by immunofluorescence double staining. To evaluate the protective effects of TQHXD-containing cerebrospinal fluid (CSF) on BMECs injured by oxygen–glucose deprivation and reperfusion, cell survival rate was determined using the CCK-8 assay and cell apoptosis was determined by fluorescein isothiocyanate (FITC)-Annexin V/PI. Autophagy was detected using transmission electron microscopy. ResultsThe results showed that TQHXD-CSF significantly ameliorated oxygen-glucose deprivation/reperfusion (OGD/R)-induced injury in BMECs. Confocal microscopy and Western blot results showed that TQHXD-CSF reduced autophagy-related protein expression and autophagosome number. The results of the western blotting indicated that TQHXD-CSF caused a marked increase in the phosphorylation of protein kinase B and phosphoinsotide-3 kinase (Akt/p-Akt and PI3K/p-PI3K, respectively) and their expression levels were down-regulated after treatment with pathway inhibitor, ZSTK474. Furthermore, in a MCAO model in rats, TQHXD markedly increased p-PI3K, p-Akt and p-mTOR, whereas the autophagy related proteins decreased. ConclusionsTaken together, these findings demonstrate that TQHXD protects against ischemic insult by inhibiting autophagy through the regulation of the PI3K/Akt/mammalian target of rapamycin (mTOR) pathway and that TQHXD may have therapeutic value for protecting BMECs from cerebral ischemia.

Quality Evaluation of Market Acacia catechu by Fingerprint-Chemical Pattern Recognition

Acacia catechu (L.f.) Willd, a leguminous plant, is included in the 2020 edition of the Chinese Pharmacopoeia and is mainly used to treat eczema, mouth ulcers, diarrhea, bruising, and traumatic hemorrhage. However, there are imported and domestic Acacia catechu samples available in China, and their quality and price are very different, which seriously affects the safety and stability of their clinical application. Importantly, there is no simple and effective method for identifying or classifying grades of Acacia catechu. In this study, 47 batches of commercial Acacia catechu were used for identifying or classifying grades of Acacia catechu using high performance liquid chromatography (HPLC) combined with chemometric analysis. Firstly, gradient elution was adopted with 0.05% phosphoric acid water (A)-methanol (B) as the mobile phase to establish chromatographic conditions. The HPLC chromatograms of 47 batches of Acacia catechu samples were analyzed by the “Similarity Evaluation System for Chromatographic Fingerprint of TCM” software (version 2012A). The common peaks of Acacia catechu were identified to evaluate the similarity. Based on the determination results of fingerprint chromatographic peak area, the quality of the collected Acacia catechu was evaluated by chemometric methods such as CA, PCA, and OPLS-DA. The results showed that the collected Acacia catechu samples were significantly divided into three categories. The first-class samples were all imported Acacia catechu except S9 sample, which was domestic Acacia catechu; the second-class samples were partly domestic Acacia catechu and partly imported Acacia catechu; and the third-class samples were all domestic Acacia catechu. Moreover, OPLS-DA of 47 batches of samples showed that the contents of catechin and the total contents of catechin and epicatechin could be used as key indicators for assessing the quality of Acacia catechu. The developed HPLC fingerprint and quantitative analysis method of multi-indicator components can be used for classification and quality evaluation of market Acacia catechu, which has a significant reference value for developing Acacia catechu grade quality standards.

Characterization of quality differences of Ophiopogonis Radix from different origins by TLC, HPLC, UHPLC-MS and multivariate statistical analyses

Ophiopogonis Radix is normally used as a traditional medicine and healthy food in China. Thin-layer chromatography (TLC), total saponin content analysis, high-performance liquid chromatographic (HPLC) fingerprinting, ultra-high-performance liquid chromatographic (UHPLC) tandem mass (MS) spectra and multivariate statistical analyses were used to analyze differences in quality of Ophiopogonis Radix from three main producing areas in China. The TLC spots of samples obtained from Zhejiang were clearer and darker than samples from Sichuan and Shandong. Additionally, HPLC fingerprinting analysis of 14 common peaks of the Ophiopogonis Radix samples indicated that the similarities of the 15 batches of samples were between 0.94 and 0.99. Finally, UPLC-Orbitrap MS analysis was used to identify the 13 common peaks of Ophiopogonis Radix. These results showed that Ophiopogonis Radix obtained from different regions can be distinguished well using a combination of analytical techniques, based on differences in their chemical components. The combination of these techniques allows comprehensive and reliable quality evaluation of Ophiopogonis Radix. This study provides references for determining the quality of Ophiopogonis Radix obtained from different regions, which will guide its use as medicine or food.

Spectrum-effect relationship between HPLC fingerprint and antioxidant of “San-Bai Decoction” extracts

“San-Bai Decoction” (SBD) has been a traditional Chinese medicine compound preparation for replenishing Qi and promoting blood circulation, whitening skin, and removing blemishes since ancient times. However, its chemical composition and antioxidant activity are not clear thus far, which limits the in-depth study on its pharmacodynamic material basis and efficacy. The objective of this study was to establish the fingerprint profile of SBD, assess its antioxidant activity by measuring 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging ability, and find the spectrum-effect relationship of SBD by Grey Relation Analysis (GRA) and Partial Least Squares Regression (PLS). In this study, the fingerprint of SBD was established by high performance liquid chromatography (HPLC), and 20 common peaks were found, among which 6 peaks were designated. The similarities between the fingerprints of 12 batches of SBD and the reference fingerprint (R) were all greater than 0.900. Meanwhile, the antioxidant activities of all batches were concentration-dependent in their linear regression equation. The result of GRA showed that the correlation order of 20 common peaks for DPPH radical scavenging was X13 > X7 > X3 > X6 > X10 > X11 > X4 > X12 > X2 > X18 > X9 > X5 > X19 > X1 > X20 > X16 > X17 > X15 > X8 > X14. At the same time, PLS study demonstrated that the contribution of six identified characteristic peaks to DPPH radical scavenging ability was X1 = X7 > X6 > X19 > X20 > X16. In this study, the spectrum-effect relationship of SBD between its HPLC fingerprint and the antioxidant activity can be used to screen the pharmacodynamic substance basis of its antioxidant action and lay the foundation for establishing quality standards and product development.

Active Compounds and Targets of Yuanzhi Powder in Treating Alzheimer's Disease and Its Relationship with Immune Infiltration Based on HPLC Fingerprint and Network Pharmacology.

Background Yuanzhi powder (YZP) has been extensively investigated as a natural prescription with therapeutic benefits for Alzheimer's disease (AD). However, its active compounds and underlying immune mechanism for treating AD are still unclear. This study aimed to investigate the immune mechanism of YZP against AD through high-performance liquid chromatography (HPLC)-based network pharmacology and gene chip technology. Methods Active components of YZP were obtained from HPLC and public databases. Subsequently, GSE5281, GSE28146, GSE29378, and GSE97760 from the Gene Expression Omnibus (GEO) database were downloaded to extract AD difference genes (DEGs). The active components-targets network and protein interaction network were then constructed by Cytoscape. The biological processes and signaling pathways, which implicate the targets of YZP for AD, were analyzed using the ClueGo Cytoscape plug-in. Molecular docking experiments were performed to verify the affinity of targets and ligands. Ultimately, the link between the hub genes and immune cell infiltration was assessed via CIBERSORT. Results 83 YZP active compounds and 641 DEGs associated with AD, including quercetin, berberine, 3,6′-disinapoylsucrose, coptisine, and palmatine, were evaluated. We showed that FOS, CCL2, and GJA1 were the core targets and that the gap junction is an essential signaling pathway in YZP for AD. Furthermore, the AD group had a higher infiltration level of naïve B cells and resting CD4 memory T cells, as determined by the CIBERSORT. Notably, the immune cells-targets network demonstrates that GJA1 and GRM1 are intimately related to naïve B cells and plasma cells. Conclusions YZP may help treat AD by targeting proteins with key active compounds to regulate naïve B cells and plasma cells. Our results demonstrate a new immune mechanism for treating AD with YZP.

Quality Markers of Dendrobium officinale by "Oligosaccharide-Spectrum-Effect" Relationships.

Dendrobium officinale Kimura et Migo has been used as a traditional Chinese medicine (TCM) and a functional food for thousands of years. Carbohydrate is one of the most important effective substances and indicative components in D. officinale. However, since the qualitative and quantitative analysis of polysaccharides in D. officinale remains a challenge and limitation, herein, an oligosaccharide-quality marker approach was newly developed for quality assessment of D. officinale by spectrum–effect relationships between high performance liquid chromatographic (HPLC) fingerprints and anti-inflammatory effects. The HPLC fingerprints of 48 batches of oligosaccharides from D. officinale (DOOS) were developed and analyzed with similarity analysis (SA) and hierarchical cluster analysis (HCA), and eight common peaks were identified. In vitro screening experiment indicated that DOOS potentially inhibited nitric oxide (NO) production and effectively reduced the release of inflammatory cytokines, such as TNF-α, IL-6, and IL-1β in RAW 264.7 cells, thereby reducing the inflammatory response of cells. Finally, the HPLC fingerprint of different batches of DOOS was combined with in vitro anti-inflammatory activity to assess the spectrum–effect relationships of DOOS by gray correlation analysis (GCA), in addition, the purified oligosaccharide components were identified and validated for NO inhibitory activity. Our results showed four DOOS (maltotetraose, maltopentaose, maltohexaose, and mannohexaose) were relevant to anti-inflammatory effects and could be as quality markers for the quality control of D. officinale. It suggests that the “oligosaccharide-spectrum-effect” relationships approach is a simple and reliable method for the quality control of herb medicines or nutritious foods.

Multiple Fingerprint Profiles and Chemometrics Analysis of Polysaccharides From the Roots of Glehnia littoralis

The quality of polysaccharides from different regions was studied by using multiple fingerprint analysis and chemometric analysis. Polysaccharides from 10 batches of Glehnia littoralis were compared based on Fourier-transform infrared spectroscopy (FT-IR), high-performance liquid chromatography (HPLC), gel permeation chromatography (GPC), and proton nuclear magnetic resonance (1H-NMR). According to the results, the 10 batches of polysaccharides from G littoralis had high similarity by analyzing HPLC, FT-IR, 1H-NMR, and GPC fingerprints. Through cluster analysis, samples and adulterants in different regions could be classified. Three monosaccharides (galactose, glucose, and galacturonic acid), molecular weights (4.33 × 105-4.91 × 105, 4.04 × 104-4.71 × 104, and 5.02 × 103-5.83 × 103), and H-1 (4.99, 5.39, and 5.42 ppm) of α-glucose could be used as markers for quality control of medicinal materials of the roots of G littoralis.

Study on the Spectrum-Effect Correlation of Anti-Inflammatory Active Extract of Sauropus spatulifolius Beille.

Sauropus spatulifolius Beille (S. spatulifolius) is a commonly used medicine of the Bourau and Yao nationalities. However, the composition of S. spatulifolius is complex, and simple chemical fingerprints cannot accurately evaluate the relationship between its composition and efficacy. In this study, high-performance liquid chromatography (HPLC) method was used to establish the fingerprint of the ethyl acetate extract of S. spatulifolius. Based on the evaluation of the similarity of chromatographic fingerprints of traditional Chinese medicine, combined with cluster analysis and principal component analysis (PCA), the common peaks of fingerprints were evaluated. The anti-inflammatory effect data were extracted through the dimethylbenzene-induced ear-swelling model in mice. The gray relational analysis (GRA) combined with partial least squares regression (PLSR) was used to study the spectrum-effect correlation of S. spatulifolius. As a result, the HPLC fingerprint of the ethyl acetate extract of S. spatulifolius was established, and 18 common peaks were identified. Except for S6, the other similarities are all above 0.915. The reference substance control method was used to identify two absorption peaks, namely, protocatechuic acid and caffeic acid. The cluster analysis results showed that 10 samples from different origins were grouped into four categories, which was consistent with the PCA results. Ethyl acetate extract of 10 batches of S. spatulifolius could significantly inhibit the ear swelling of mice (P < 0.01). Through GRA, the order of the contribution of each chemical component to the anti-inflammatory efficacy was obtained. The results of PLSR showed that the VIP values of peaks 3, 4, and 12 were greater than 1 and were positively correlated with anti-inflammatory activity. In this study, the HPLC fingerprint of the ethyl acetate extract of S. spatulifolius was established. Through the study of the spectrum-effect correlation, the anti-inflammatory active substance of the ethyl acetate extract of S. spatulifolius was obtained. The anti-inflammatory effect of S. spatulifolius was the result of the joint action of multiple ingredients. This research helps to quickly and accurately discover the active ingredient groups of traditional Chinese medicine and provides new ideas and methods for studying the effective substances of traditional Chinese medicine.

Interbatch quality control of the extract from Artemisia frigida Willd. by spectrum-effect relationship between HPLC fingerprints and the total antioxidant capacity

ABSTRACT Similarity analysis of complicated chromatographic profiles is a potential protocol for the authentication and quality control of traditional Chinese medicine to guarantee prospective biological activity. In this work, a high-performance liquid chromatography (HPLC) method was established to identify and control the inter-assay quality of extract from Artemisia frigida Willd. Meanwhile, a partial least squares regression (PLSR) was applied to construct a spectrum–effect relationship between the HPLC fingerprints and the total antioxidant capacity of the extract from Artemisia frigida Willd. The similarity correlation coefficient of 10 batches of extract was ≥0.948 could guarantee the 10 batches of showed the inter-assay stability of antioxidant capacity of extract from Artemisia frigida Willd. Moreover, PLSR analysis showed that four efficacy-related components were closely related to antioxidant effect. Therefore, this study provided a flexible and efficient method for the quality and the stable biological activity assessment of Artemisia frigida Willd.

Quality Status Analysis and Intrinsic Connection Research of Growing place, Morphological Characteristics, and Quality of Chinese Medicine: Cyperi Rhizoma (Xiangfu) as a Case Study.

Materials and Methods The macroscopic characteristics of CR as well as its moisture, ethanol extract, essential oil, total ash, and acid-insoluble ash contents were examined and calculated. In addition, qualitative identification and quantitative determination of α-cyperone, cyperotundone, and nootkatone were simultaneously performed, and a high-performance liquid chromatography (HPLC) fingerprint chromatogram was established. Inductively coupled plasma mass spectrometry and gas chromatography methods recorded in ChP were used to measure the contents of residues of heavy metal and deleterious elements as well as residues of organochlorine pesticide, respectively. Hierarchical cluster analysis and typical canonical correlation analysis were performed using Origin 9.1 and SPSS 23.0 to explore the correlation between CR's growth area, morphological characteristics, and quality. Results Of the 47 batches of CR analyzed, only 4 collected from the province of Shandong had a flat appearance, which did not accord with the macroscopic characteristics of CR. Overall, only 4 batches met ChP standards for CR. In addition, 30 and 38 batches did not meet the requirements for moisture content and essential oil content, respectively. The similarity values of HPLC fingerprints ranged from 0.568 to 0.986. Results of hierarchical cluster analysis for ethanol extracts, essential oil, α-cyperone, cyperotundone, and nootkatone and the HPLC fingerprints (total peak time and peak area) suggested that the samples could be classified into four clusters, with no significant difference in growth geographic areas among them. Results of canonical correlation analysis indicated that the first canonical pair could represent the correlation between macroscopic characteristics (vector 1) and chemical quality (vector 2), with shorter diameter and length denoting lower ethanol extract content and higher nootkatone content in a single grain of CR. Conclusions Crude medicinal materials were collected and examined in this study to reflect the overall quality status of CR in China. The methods chosen to detect, calculate, and analyze the quality of CR were suitable to the investigation, and the results are crucial not only for estimating the current quality status of CR, but also for conducting further research into its cultivation, quality assurance, and commodity specification. Besides, this mode of investigation could be used to evaluate other medicines.