Quantitative Analysis Of Multi-components Research Articles

The Development and Validation of Simultaneous Multi-Component Quantitative Analysis via HPLC–PDA Detection of 12 Secondary Metabolites Isolated from Drynariae Rhizoma

Drynariae Rhizoma (DR) is a functional food and traditional medicine that has been widely used for bone and joint disorders for thousands of years. In this study, 14 compounds were isolated from DR, and their structures were identified using UPLC/QTOF–MS, UPLC–ESI/LTQ–Orbitrap–HRMS, and 2D NMR and compared with those obtained in previous studies. An HPLC–PDA multi-component simultaneous quantitative determination method was developed for 12 of the 14 DR-derived compounds, excluding compounds with a content <1.5 mg. The developed HPLC method was validated based on linearity (r2 ≥ 0.999), limit of detection (0.01–0.65 μg/mL), limit of quantification (0.04–1.97 μg/mL), intra-day precision and accuracy ranges (0.06–2.85% and 95.03–104.75%, respectively), and inter-day precision and accuracy ranges (0.24–2.83% and 95.75–105.75%, respectively). The developed analysis method improved the resolution of compounds 4 and 5. In addition, this is the first quantitative analysis of compounds 7, 8, and 11 and the first simultaneous quantitative analysis of 12 compounds, including compounds 4, 7, 8, 10, 11, and 14. This study developed a rapid, accurate, and economical HPLC method for performing the simultaneous quantitative analysis of 12 secondary metabolites isolated from DR.

Integrated quality control strategy for red yeast rice combining DSC and HPLC profiles

Red yeast rice (RYR) is a popular food product with significant quality control challenges. This study first established a gradient elution tower plate theory suitable for gradient elution and proposed a dual-label system calibration method to monitor and correct chromatographic systems for changes. Subsequently, high-performance liquid chromatography as the analytical method, and a five-wavelength maximization profile was employed to obtain comprehensive chromatographic information on RYR. A novel multi-marker assay by monolinear method (MAML) was employed for the accurate quantitative determination of quality markers. A differential scanning calorimetry (DSC) quantized fingerprint was established to study complete thermal changes. The comprehensive linear quantitative fingerprint method (CLQFM) was also developed to evaluate fingerprints. Results showed that the CLQFM divides the samples into four quality grades, MAML yielded better results than quantitative analysis of multi-components by a single marker method. Thermal analysis revealed that treatment temperatures higher than 193.7 °C for extended periods should be avoided to maintain sample quality. This study promotes the dual control of the quality and thermal stability of RYR, providing a new direction for food control.

Integration of fingerprint-activity relationship and chemometric analysis to accurately screen Q-markers for the quality control of traditional Chinese medicine compound preparation: Jinlian Qingre granules as an example

This study aims to develop a comprehensive strategy that accurately screens and quantifies quality markers (Q-markers) by combining LC-MS metabolomics with fingerprint-effect relationships, chemometrics, and QAMS methods for the quality control in traditional Chinese medicine compound preparations (CMP). First, the chemical compositions in Jinlian Qingre granules (JQG) were analyzed and identified using ultrahigh-performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS). Next, the HPLC fingerprint of JQG was established, and chemometrics were employed to evaluate JQG quality. Then, potential anti-inflammatory bioactive constituents were screened using a fingerprinting-effect relationship model utilizing partial least squares regression (PLSR) and a back propagation neural network optimized using a genetic algorithm (GA-BPNN). On this basis, Q-markers of JQG were determined and analyzed using quantitative analysis of multi-components by the single marker (QAMS) employing three components as reference substances (RS). 24 chemical constituents in JQG were characterized using UHPLC-HRMS. Twenty-four common peaks were assigned to the fingerprint. Six compounds, mangiferin, 2′'-O-beta-L-galactopyranosylorientin, orientin, veratric acid, vitexin, and harpagoside, identified from the common peaks associated with anti-inflammatory efficacy, could be used as Q-markers of JQG. The content determination results displayed no significant difference in the content of Q-markers in 20 batches of JQG samples measured using the external standard method and QAMS. Q-markers could be efficiently screened and determined by integrating multidisciplinary technologies. This comprehensive strategy could be beneficial for the thorough evaluation of CMP.

A quality evaluation method of lotus leaf based on its lipid lowering components using QAMS and chemometrics

IntroductionLotus leaf has long been used as food and medicine in China and is well-known for its lipid-lowering effects. However, there is a lack of a comprehensive quality evaluation for lotus leaf due to the absence of consideration of the correlation between various components and their efficacy. ObjectivesThis study aims to find out the key bioactive components that can be used for quality evaluation of lotus leaf on lipid-lowering effect. MethodsThirteen compounds were characterized in the lotus leaf using ultra-high- performance liquid chromatography-time-of-fight mass spectrometry (UPLC-Q-TOF-MS). Five alkaloids and four flavonoids were identified according to their lipid-lowering activities reported in literatures. Then, the contents of these nine components were analyzed in 39 batches of lotus leaves growing in different locations using high performance liquid chromatography diode-array detector (HPLC-DAD), and further evaluated by quantitative analysis of multi-components by single marker (QAMS) and chemometrics. The anti-adipogenic activity of lotus leaves were evaluated for their inhibitory effect on the PPARγ expression by luciferase assay. ResultsThe 39 batches were clustered into two regions, the north and the south, based on the contents of these components. Three alkaloids, nuciferine, N-nornuciferine, and asimilobine, and three flavonoids, astragalin, hyperoside, and trifolioside, were found to serve as the key factors behind the region differences. Their contents were higher in Guangchang County of Jiangxi Province than other habitat locations. Moreover, the luciferase assay combined with chemometrics showed that these components were positively correlated with lipid-lowering activity of lotus leaf. ConclusionsThree alkaloids and three flavonoids were screened out and could be used as key compounds for quality evaluation of lotus leaf on lipid-lowering effect.

A multi-strategy platform for Q-markers screening and quality control of Wuzi Yanzong Wan based on fingerprint and network pharmacology

Wuzi Yanzong Wan (WZYZW) is a classic formula for tonifying the kidneys in Traditional chinese medicine (TCM) and is widely used clinically in diseases of the reproductive system, immune system and nervous system. Therefore, the quality control and prediction quality markers (Q-markers) of WZYZW is of positive significance.In this study, We first established the fingerprints, calibrated 28 common peaks, identified 12 common peaks of gallic acid, geniposidic acid, chlorogenic acid, ellagic acid, rutin, hyperoside, verbascoside, astragalin, kaempferol, schisandrol A, schisandrol B and schisandrin B, and the similarity was greater than 0.9 of 10 batches.Then, withing network pharmacology, we screened 41 core targets such as PIK3CA, AKT1, PIK3CB, potential pathways such as pathways in cancer and PI3K-Akt signalling pathway of 12 components. The results showed that they were tightly connected to predicated targets and pathways. Finally, by using the hyperoside as internal reference standard, a Quantitative analysis of multi-components (QAMS) for WZYZW was established to determine the contents of WZYZW. The percent difference(%) showed that there was no significant difference between the quantitative results of external standard method (ESM) and QAMS method. Contents of the 11 components above were 0.0091 ∼ 0.0185 %, 0.1019 ∼ 0.1361 %, 0.0518 ∼ 0.0718 %, 0.0130 ∼ 0.0320 %, 0.0084 ∼ 0.0190 %, 0.0621 ∼ 0.1192 %, 0.0127 ∼ 0.0419 %, 0.0006 ∼ 0.0089 %, 0.0212 ∼ 0.0261 %, 0.0061 ∼ 0.0400 %, 0.0084 ∼ 0.0103 %, respectively. Overall, the method developed was applied to the quality evaluation and Q-markers research of WZYZW as well as provided guiding significance for the Q-marker research of TCM.

Complete composition analysis of polysaccharides based on HPAEC-PAD coupled with quantitative analysis of multi-components by single marker.

Monosaccharide compositions analysis (MCA) is indispensable for structural characterisations and structure-activity relationships of plant polysaccharides. To develop a concise and direct MCA method, we established a quantitative analysis of the multi-monosaccharaides by single marker (QAMS) by high-performance anion-exchange chromatography with pulsed-amperometric detection (HPAEC-PAD) method. A stable and reproducible HPAEC-PAD method for simultaneous determination of aldoses, ketoses and uronic acids (i.e., l-arabinose, d-xylose, d-ribose, l-rhamnose, d-fucose, d-mannose, d-glucose, d-galactose, d-fructose, d-glucuronic acid and d-galacturonic acid) was established by systematic optimisation of stationary phases, column temperatures and elution programmes. On this basis, the QAMS method was proposed through comprehensive investigations of relative correction factor (RCF) variations under different influencing factors, for example, sample concentrations, flow rates, and column temperatures. Using rhamnose as an internal reference standard, the contents of the other monosaccharidecomponents in polysaccharides from Panax quinquefolium L. and Achyranthes bidentata Bl. samples were simultaneously determined by QAMS, and there was no significant difference between the results from the QAMS and external standard method (t test, P > 0.520). In addition, a MCA fingerprinting of 30 batches of P. quinquefolium polysaccharide was established by HPAEC-PAD, and six common peaks were assigned and determined. The established HPAEC-PAD-QAMS method was successfully applied to the MCA of polysaccharides from P. quinquefolium and A. bidentata after optimisation of hydrolysis conditions. HPAEC-PAD-QAMS was proposed and established for MCA of plant polysaccharides for the first time.

Quality Evaluation of Sanguisorbae Radix via Python Aided Optimization Fingerprint Chromatography Combined with Quantitative Analysis of Multi-components by Single Marker

Quality Evaluation of Sanguisorbae Radix via Python Aided Optimization Fingerprint Chromatography Combined with Quantitative Analysis of Multi-components by Single Marker

Multicomponent Analysis of Liuwei Dihuang Pills by a Single Marker Quantification Method and Chemometric Discrimination of Fingerprints.

An effective and comprehensive quality evaluation method for Liuwei Dihuang pills (LDP) was established by the simultaneous determination of 8 active components in LDP by the quantitative analysis of multicomponents by single marker (QAMS) method and high-performance liquid chromatography (HPLC) fingerprint combined with chemometrics. These 8 active components were determined by QAMS and the external standard method (ESM), and the quantitative results of the two methods were compared to validate the accuracy and feasibility of the QAMS method. 8 active components showed good linear relationships within their ranges, whose average recoveries were 99.7∼102.3%. No significant difference was found (P > 0.05) in the quantitative results determined by QAMS and ESM. Furthermore, the fingerprint of LDP was also established, with 11 common peaks identified, and the similarity of the fingerprints of 21 batches of LDP was greater than 0.95. The 21 batches of LDP were basically divided into 3 groups by hierarchical cluster analysis (HCA) and principal component analysis (PCA), and 3 differential markers were screened out by orthogonal partial least squares discriminant analysis (OPLS-DA). The established QAMS method is accurate, economical, fast, and convenient and can simultaneously determine the content of 8 active components in LDP. HPLC fingerprint combined with chemometric analysis more comprehensively evaluated the quality consistency of different batches of LDP and analyzed the markers that cause quality differences between batches. It can provide a scientific basis and reference of quality consistency evaluation for the manufacturers and drug regulatory departments of the preparation.

Quality Evaluation of Volatile Oil in Yanyangke Mixture (YM) Based on GC-MS Fingerprint, GC Multicomponent Quantitative Analysis and Chemical Pattern Recognition Analysis.

Yanyangke mixture (YM) is composed of 12 kinds of traditional Chinese medicine (TCM) used for the treatment of patients with cough, dry throat and other diseases caused by acute or chronic pharyngitis or patients with difficulty in expectoration. With the wide application of YM in clinical practice, its quality control has attracted huge attention. Based on the multi-component characteristics of Chinese herbal medicines, it is pertinent to establish a quality evaluation system. A new idea is to adopt gas chromatography-mass spectrometry (GC-MS) chemical composition identification, GC-MS fingerprint, and GC content determination as a potential quality control index of the volatile oil in YM. In this study, the volatile oil of YM was extracted by steam distillation, and the chemical components of the volatile oil were analyzed by GC-MS, and 43 chemical components were identified. The fingerprint of the volatile oil from YM was established and the similarity evaluation was performed. Combined with chemometric methods, such as cluster analysis, principal component analysis and partial least squares analysis, the chemical composition differences of the volatile oil from different batches of YM were compared and the symbolic components affecting the quality of the volatile oil from different batches of YM were excavated. Finally, three components were selected as the potential active component markers of YM and the GC content determination method of these three components was established. A rapid, reasonable, and effective quality evaluation and control method of YM volatile oil was established, which provided a reference for further development and research on YM, as well as a new idea for research on other TCM prescriptions.

Quality evaluation of Compound Cheqian Tablets based on UPLC-Q-TOF-MS/MS, network pharmacology and "double external standards" QAMS

The Compound Cheqian Tablets are derived from Cheqian Power in Comprehensive Recording of Divine Assistance, and they are made by modern technology with the combination of Plantago asiatica and Coptis chinensis. To investigate the material basis of Compound Cheqian Tablets in the treatment of diabetic nephropathy, in this study, the chemical components of Compound Cheqian Tablets were characterized and analyzed by UPLC-Q-TOF-MS/MS, and a total of 48 chemical components were identified. The identified chemical compounds were analyzed by network pharmacology. By validating with previous literature, six bioactive compounds including acteoside, isoacteoside, coptisine, magnoflorine, palmatine, and berberine were confirmed as the index components for qua-lity evaluation. Furthermore, the content of the six components in the Compound Cheqian Tablets was determined by the "double external standards" quantitative analysis of multi-components by single marker(QAMS), and the relative correction factor of isoacteoside was calculated as 1.118 by using acteoside as the control; the relative correction factors of magnoflorine, palmatine, and berberine were calculated as 0.729, 1.065, and 1.126, respectively, by using coptisine as the control, indicating that the established method had excellent stability under different conditions. The results obtained by the "double external standards" QAMS approximated those obtained by the external standard method. This study qualitatively characterized the chemical components in the Compound Cheqian Tablets by applying UPLC-Q-TOF-MS/MS and screened the pharmacodynamic substance basis for the treatment of diabetic nephropathy via network pharmacology, and primary pharmacodynamic substance groups were quantitatively analyzed by the "double external stan-dards" QAMS method, which provided a scientific basis for clarifying the pharmacodynamic substance basis and quality control of Compound Cheqian Tablets.

Quality Evaluation of Bioactive Ingredients in Lianhua Qingwen Capsule Based on Quantitative Analysis of Multicomponent by the Single Marker Method and the Chemical Recognition Patterns Method

Based on high performance liquid chromatography with the diode array detector (HPLC-DAD), a new strategy for simultaneous determination of ten bioactive ingredients in Lianhua Qingwen capsule (LHQW) was developed for comprehensive quality assessment of LHQW. In this work, with rhein regarded as the internal reference substance (IRS), the relative correction factors (RCFs) of neochlorogenic acid, amygdalin, chlorogenic acid, forsythoside A, quercitrin, phillyin, glycyrrhizic acid, isoforsythiaside, and (+) pinoresinol-β-D-glucoside were calculated for simultaneous determination of ten bioactive ingredients. More importantly, compared to previous work, the simultaneous determination of the content of ten pharmacologically important active ingredients at one detection wavelength with only one reference substance has been achieved. Based on the contents of ten bioactive ingredients, the quality of the 20 batches of LHQW samples was further analyzed by chemical recognition patterns method. Ten bioactive ingredients showed a good linear relationship in their respective concentration ranges (r ≥ 0.999). The relative standard deviations (RSDs) of precision (≤4.62%), stability (≤4.04%), repeatability (≤3.87%), and the average recovery of ten bioactive components (99.8%∼104.1%) demonstrated the QAMS developed for LHQW which had good durability. The correlation coefficient ( P > 0.05 ) showed that no significant difference existed in the results of QAMS and external standard method (ESM). Hierarchical clustering analysis (HCA) divided samples into three main groups. Radar plot analysis and principal component analysis (PCA) found some quality differences existed between the three groups of samples. Orthogonal partial least-squares discrimination analysis (OPLS-DA) showed that forsythoside A could be used as the primary marker responsible for the quality differences. In conclusion, the established QAMS method combined with chemometric analysis can simultaneously determine the content of 10 active components and comprehensively evaluate the quality of different batches of LHQW. It can provide scientific basis and reference of quality consistency evaluation for the formulation manufacturers and drug regulatory authorities.

Quality evaluation of Keteling capsules based on fingerprinting, multicomponent quantification, and quantitative prediction

The Keteling capsule (KC) is a traditional Chinese medicine (TCM) made from the dried extract of Ficus microphylla and an appropriate amount of chlorpheniramine maleate. It is widely used to treat cough and relieve asthma. Despite its extensive usage, a rapid and comprehensive quality evaluation strategy for KC remains a challenge. This study introduces an electrochemical fingerprint analysis technique, in addition to the commonly employed HPLC fingerprints, for efficient and convenient quality evaluation. Moreover, a cost-effective, rapid, and accurate multi-component quantification technique known as the “Multi-markers assay by the monolinear method (MAML)” and the “FT-IR quantitative model” were explored. The HPLC fingerprints were evaluated using a systematically quantified fingerprint method, while the electrochemical fingerprints, based on the Belousov-Zhabotinsky oscillation reaction principle, were effectively analyzed and characterized using oxidation induction times and oscillation lifetimes. Multi-component quantitative analysis was carried out through the MAML and FT-IR quantitative models. The HPLC fingerprint successfully classified the 22 samples into eight grades with excellent discrimination. Active ingredient content analysis was achieved using reliable parameters obtained from electrochemical fingerprinting. The no significant difference in the quantitative results proves the accuracy of the MAML method. Additionally, successful FT-IR quantitative prediction models were developed for chlorogenic acid, isovitexin, and chlorpheniramine maleate. This study offers a dependable and effective approach for enhancing the quality control of KC, and it can provide new insights for improving the quality analysis methods in the field of TCM.

Multi-component content determination of Dracocephalum tanguticum by quantitative analysis of multi-components by single-marker

This study aims to establish a method for the simultaneous determination of 7 active components in Dracocephalum tanguticum and to evaluate the quality of medicinal materials from different habitats. The method was established with high performance liquid chromatography(HPLC) and the gradient elution was performed with the mobile phase of acetonitrile-methanol-0.2% phosphoric acid solution at a column temperature of 35 ℃, an injection volume of 15 μL, and a flow rate of 0.6 mL·min~(-1). The detection wavelength was set as 215 nm. With rosmarinic acid as the internal reference, the relative correction factors and the content of other 6 components were calculated. The results were compared with those obtained with the external standard method. The results showed that the samples from Huangzhong county, Qinghai province had the best quality, with the highest content of p-hydroxybenzoic acid, cosmosiin, rosmarinic acid, oleanolic acid, and ursolic acid(9.29, 12.14, 6.02, 3.11, 17.67 mg·g~(-1) respectively). The samples from Chaya county, Tibet autonomous region ranked the second, with the highest content of betulin and betulinic acid(15.53, 7.17 mg·g~(-1), respectively). The method is accurate, reliable, and repeatable and suitable for the simultaneous determination of multiple components in D. tanguticum. The content of functional components varied in the samples from different producing areas and can be used as the indicator for the quality evaluation of medicinal materials.

Determination of Ten Flavonoids in the Raw and Fermented Fructus Aurantii by Quantitative Analysis of Multicomponents via a Single Marker (QAMS) Based on UPLC.

Fermented Fructus Aurantii (FFA) is widely used in South China for the treatment of functional dyspepsia. Naringin, neohesperidin, and other flavonoids are the main pharmacodynamic components of FFA. A new method is presented for the simultaneous determination of 10 flavonoids (including flavonoid glycosides and aglycones) in FFA using the quantitative analysis of multicomponents via a single marker (QAMS) approach and is used to investigate changes in flavonoids during fermentation. The viability and precision of QAMS were validated against the ultrahigh-performance liquid chromatography (UPLC), with various UPLC instruments and chromatographic conditions being evaluated. Differences between raw Fructus Aurantii (RFA) and FFA were examined using orthogonal partial least squares discrimination analysis (OPLS-DA) and content determination. The influence of various fermentation conditions on flavonoids was also investigated. There were no appreciable differences between the QAMS and the external standard method (ESM), demonstrating that QAMS is an improved method for the determination of FA and FFA. FFA and RFA can be readily distinguished based on OPLS-DA chemometric modelling and the corresponding chromatograms. In addition, the flavonoid changes after fermentation. Fermentation considerably reduced the contents of flavonoid glycosides, while increasing hesperidin-7-O-glucoside and flavonoid aglycones. Moreover, fermentation conditions impact multiple flavonoids in FA, so controlling these conditions is necessary for the quality control of fermented FA products. This QAMS approach is useful for detecting numerous components in RFA and FFA simply, quickly, and efficiently, thus strengthening the quality control of FA and its fermented products.

Comprehensive quality evaluation of Qizhi Xiangfu Pills based on Quantitative Analysis of Multi-components by Single Marker combining with GC Fingerprint and Chemometrics.

Qizhi Xiangfu Pills (QXPs) is a traditional Chinese medicine (TCM) used clinically for Qi stagnation and blood stasis. The current quality control of QXPs in the ministry standards and the reported literature is minimal, and requires to be improved. This study aimed to analyze and determine the active ingredients in QXPs for its overall evaluation. In this study, a quantitative analysis of multi-components by single marker (QAMS) method was established to simultaneously determine the caryophyllene oxide, cyperotundone, ligustilide and α-cyperone in QXPs by GC. Moreover, the GC fingerprints of 22 batches samples were also established, and the common peaks were initially identified by GC-MS, then classified in various dimensions using chemometric methods, and the main markers causing the discrepancies between groups were analyzed by orthogonal partial least squares discrimination analysis (OPLS-DA). Compared with internal standard method (ISM), the determination results obtained by QAMS had no significant difference. 22 common peaks were distinguished in the fingerprint of 22 batches QXPs, 17 of which were identified, and the similarity of the fingerprint was greater than 0.898. The 22 batches of QXPs were roughly divided into 3 categories, and 12 main markers causing the discrepancy were discovered. The established QAMS combined with GC fingerprint and chemometrics method is convenient and feasible, which helps to improve the quality evaluation of QXPs and provides a demonstration for the relative study of compound preparations and single herbs. A quantitative analysis of multi-components by single marker combining with GC fingerprint and chemometrics method was established to evaluation the quality of Qizhi Xiangfu Pills for the first time.

A field trials-based study reveals the effects of chlormequat chloride on the quality of medicinal plant Salvia miltiorrhiza Bge. (Lamiaceae) in practical cultivation

The application of some specific plant growth regulators in the cultivation of herb medicines for yield promotion is a widespread phenomenon at present. But most of these agrochemical uses in medicinal plants are without effect verification and are illegal according to related regulations, thus it is necessary to conduct studies about the effect evaluation of these plant growth regulators on medicinal plants in actual production conditions. In this study, according to a preliminary investigation that chlormequat chloride (CCC) is widely used in the cultivation of the common herb Salvia miltiorrhiza, the effects of CCC on the quality of S. miltiorrhiza were comprehensively evaluated with both metabolomics-based UPLC/Q-TOF MS method and multi-component quantitative analysis based on samples from two years field trials. A global chemical constituent identification of S. miltiorrhiza was firstly carried out and consequently 47 tanshinones and 22 phenolics were identified or tentatively identified from the UPLC/Q-TOF MS data. Then, both the qualitative multivariate statistical analysis and quantitative determinations of major active components in S. miltiorrhiza samples that were treated with different dosages of CCC revealed that the application of CCC in S. miltiorrhiza cultivation is likely to count against the herbal quality and mainly manifested as the significant decrease of tanshinones content. Above all, this work provides a beneficial case for the recognition of the blind use of plant growth regulators in herb medicines for yield improvement and indicated that the use of PGRs in medicinal plant cultivation should be more cautious, and several further study points based on this work were discussed.

Established UPLC-MS/MS procedure for multicomponent quantitative analysis of rat plasma: Pharmacokinetics of Taohong Siwu Decoction in normal and acute blood stasis models

Ethnopharmacological relevanceAs one of China's 100 classic recipes, Taohong Siwu Decoction (THSWD) consists of Siwu Tang flavored peach kernel and safflower, and is used to nourish and activate blood. Accordingly, THSWD is mainly administered to treat blood deficiency and stasis syndrome. According to prior studies, THSWD induces antioxidant stress, inhibits inflammatory reactions, inhibits platelet aggregation, prevents fibrosis, reduces blood lipids, prolongs clotting time, prevents atherosclerosis and vascular pathology, improves hemorheological changes, and regulates related signaling pathways. Materials and methodsA sensitive analytical method was developed to detect the marker components of THSWD using UPLC-Q-TOF-MS. A rapid and sensitive UPLC-MS/MS analytical method was developed and applied to detect 16 major bioactive components in normal and acute blood stasis (ABS) rats following oral administration of THSWD. The metabolic process of THSWD in vivo was evaluated and the differences in pharmacokinetic parameters between the normal and ABS rat metabolic processes were compared. ResultsThis method was fully validated based on its excellent linearity (r2 < 0.99), satisfactory intra- and inter-day precisions (RSD <15%), and good accuracy (RE within ±14.83%). The stability, matrix effects, and extraction recoveries of the rat plasma samples were also within the acceptable limits (RSD <15%). Compared to normal rats, the pharmacokinetics of the major active constituents (except Senkyunolide G) were significantly different (P < 0.05) in the ABS model rats, indicating that the metabolism of the 16 compounds in vivo may change under disease conditions. ConclusionsIn this study, a sensitive UPLC-Q-TOF-MS method was established to analyze the main components of THSWD, and a UPLC-MS/MS analytical method was developed and applied for the pharmacokinetic parameter detection of the 16 main bioactive components in normal and ABS rats. Our findings lay the foundation for further studies on the pharmacokinetic-pharmacodynamic correlation for THSWD.

Rapid Analysis of Four Alkaloids in Uncaria rhynchophylla by Core-Shell Column HPLC and Quantitative Analysis of Multi-Components by Single Marker (QAMS)

As a traditional herbal medicine, the major alkaloids in Uncaria rhynchophylla have been proven to have blood pressure-lowering and sedative effects. It is essential to develop an effective method for the determination of the major alkaloids in U. rhynchophylla. In this research, a rapid quantitative analysis involving multi-components analysis by a single marker strategy coupled with core-shell column HPLC was adopted to analyse four alkaloids (corynoxeine, isocorynoxeine, isorhynchophylline, rhynchophylline) in U. rhynchophylla. Isorhynchophylline was selected as the internal reference substance, the content of which was determined by the traditional external standard method. Relative correction factors (RCF) between isorhynchophylline and the other three alkaloids were calculated respectively. The results showed that the QAMS method had good robustness under different HPLC instruments. Nineteen batches of U. rhynchophylla were tested. No significant difference was observed between the results by QAMS and EMS (Correlation coefficient > 0.99, p > 0.05). The QAMS method could be employed as a rapid, effective technique for the quality control of U. rhynchophylla.

Developing a novel single-marker-based method for the quantitative evaluation of the multiple active components in Corydalis yanhusuo W. T. Wang

Objective: This study was designed to develop a method for detecting differences in the chemical composition of Corydalis yanhusuo W. T. Wang using high-performance liquid chromatography with a diode array detector technology. Materials and Methods: We established a novel quantitative evaluation method for identifying multiple components in natural extracts using a single-marker method quantitative analysis of multi-components by single marker (QAMS). This method was then validated using eight alkaloid phytochemical markers designed to evaluate C. yanhusuo quality. Results: Our evaluations revealed good linearity (R2 ≥ 0.9991) within the range of tested concentrations for all eight alkaloids, with recovery ranging from 95.5% to 101.5%. The evaluations also returned stability results that fell within the acceptable range. Cluster analysis and Heatmap analyses were applied to classify and evaluate alkaloids across 21 different production areas. These results revealed a significant difference in the component profiles between samples from different origins. Conclusions: Thus, these data suggest that in the absence of a material reference, QAMS may help facilitate the stable production of C. yanhusuo. In addition, our data suggest that this method may have value as a promising alternative to common quality evaluations for controlling C. yanhusuo composition.

Modern methods for the determination of pesticide residues in beekeeping products and for the diagnostics of bee poisoning

Intensification of agricultural production is associated with the use of a significant amount of pesticides, which negatively affects the environment and human health, and food products, including beekeeping products, accordingly require mandatory control of residual amounts of pesticides. This article provides a comparative analysis of the available chromatographic methods for pesticide residue research. The necessity of using modern chromatographic methods to determine residual amounts of pesticides in samples of dead bees and beekeeping products is well-founded. Chromatographic methods of studying these indicators in different types of matrices are a priority. They are effective methods of analysis, widely used due to their versatility - they allow the analysis of complex inorganic and organic compounds in various aggregate states. But one of the most common modern methods for pesticide determination is gas and liquid three-quadrupole tandem chromatography-mass spectrometry (GC and/or LC-MS/MS). The GC-MS/MS method provides quantitative determination of analytes at a level that is an order of magnitude higher than, for example, the gas single quadrupole mass spectrometry method. Modern methods of gas and liquid chromatography in combination with quadrupoletime-of-flight mass spectrometric detection (LC/Q-TOF/MS or GC/ Q-TOF/MS) also allow qualitative and quantitative multicomponent analysis of pesticides in beekeeping products. GC and LC systems combined with high-resolution Orbitrap MS (GC-HRMS(Q-Orbitrap)/LC-HRMS (Q-Orbitrap)) have higher sensitivity, enabling ultra-trace detection, and are the most sensitive screening method for multicomponent determination of pesticide residues. Thus, the latest chromatographic methods are able to meet the needs of analytical testing and research laboratories in the field of food safety, including beekeeping products. Key words: chromatographic methods, thin-layer chromatography, gas chromatography, liquid chromatography, mass spectrometry, multi-component analysis, pesticides, honey, dead bees.