Quality Consistency Evaluation of Multiorigin Chinese Herbal Medicine by Using a Combination of Chemical Similarity and InVitro Bioequivalence: Taraxaci Herba as a Case Study.

Taraxaci Herba was derived from the dried Herba of Taraxacum mongolicum, T. borealisinense and several species from the Taraxacum genus. Taraxaci Herba has been widely used in traditional Chinese and folk medicines. According to the different growth and cultivation pattern, Taraxaci Herba could be divided into two species, wild Taraxaci Herba and cultivated Taraxaci Herba. In the present study, an accurate and reliable fingerprint approach was developed using high performance liquid chromatography(HPLC) for quality control of Taraxaci Herba. A total of 9 common peaks were marked, and the similarity of all the Taraxaci Herba samples was above 0.960. The established fingerprint method could be used for quality control of Taraxaci Herba. Furthermore, an HPLC method was established for simultaneous determination of six bioactive compounds, including monocaffeoyl tartaric acid, chlorogenic acid, caffeic acid, cichoric acid, 4,5-dicaffeoylquinic acid and luteolin in wild Taraxaci Herba and cultivated Taraxaci Herba. Moreover,chemometrics analysis such as principal component analysis and orthogonal partial least squares discriminant analysis were performed to compare and discriminate the wild samples and cultivated samples based on the quantitative data. The chemometrics results indicated that 4,5-dicaffeoylquinic acid and luteolin were significant to effectively discriminate the wild Taraxaci Herba and cultivated Taraxaci Herba samples, and these two compounds could be recognized as chemical markers for quality evaluation of wild Taraxaci Herba and cultivated Taraxaci Herba. The fingerprint analysis and quantitative analysis of multi-components could be a well-acceptable strategy for evaluation the quality of Taraxaci Herba.

Botanical drug products have batch-to-batch quality variability due to botanical raw materials and the current manufacturing process. The rational evaluation and control of product quality consistency are essential to ensure the efficacy and safety. Chromatographic fingerprinting is an important and widely used tool to characterize the chemical composition of botanical drug products. Multivariate statistical analysis has showed its efficacy and applicability in the quality evaluation of many kinds of industrial products. In this paper, the combined use of multivariate statistical analysis and chromatographic fingerprinting is presented here to evaluate batch-to-batch quality consistency of botanical drug products. A typical botanical drug product in China, Shenmai injection, was selected as the example to demonstrate the feasibility of this approach. The high-performance liquid chromatographic fingerprint data of historical batches were collected from a traditional Chinese medicine manufacturing factory. Characteristic peaks were weighted by their variability among production batches. A principal component analysis model was established after outliers were modified or removed. Multivariate (Hotelling T(2) and DModX) control charts were finally successfully applied to evaluate the quality consistency. The results suggest useful applications for a combination of multivariate statistical analysis with chromatographic fingerprinting in batch-to-batch quality consistency evaluation for the manufacture of botanical drug products.

From chemical consistency to effective consistency in precise quality discrimination of Sophora flower-bud and Sophora flower: Discovering efficacy-associated markers by fingerprint-activity relationship modeling

Unveiling the genus Taraxacum: From folk medicine to chemodiversity-driven pharmacological and toxicological outcomes-A systematic review.

Pudilan Xiaoyan Oral Liquid has been widely used in the clinical treatment of inflammatory diseases such as upper respiratory tract infections. Taraxaci Herba, as the monarch medicine in Pudilan Xiaoyan Oral Liquid, due to its multi-source, multi-origin characteristics, and the difference in the content of active ingredients in different medicinal parts, has become a potential factor for the unstable quality among different batches of Pudilan Xiaoyan Oral Liquid. In this paper, Thermo Scientific Vanquish ultra-high-performance liquid chromatography(UPLC) system was used, and the Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System(2012 Edition) issued by National Pharmacopoeia Commission was used for processing and analysis. The main common peaks were identified and contents were determined by comparison with reference substances. Fingerprints of Taraxaci Herba medicinal materials from different origins were established. 13 common peaks were identified, and 29 batches of samples from five origins had similarities above 0.90. At the same time, an ultra-high performance liquid chromatography method was developed for the determination of monocaffeoyl tartaric acid, chlorogenic acid, caffeic acid, chicoic acid, and luteolin in Taraxaci Herba. The quantitative analysis conditions were verified by methodology, and the average sample recovery was 97.30%-101.8%. The results showed that the content of the same ingredient in Taraxaci Herba from different origins and different medicinal parts was obviously different, and the fluctua-tion range was also different for different ingredients. The establishment of UPLC fingerprints for Taraxaci Herba from different regions combined with multi-component content determination methods provides a reference for improving the quality control of Taraxaci Herba medicinal materials, and also provides a source guarantee for the quality improvement of Pudilan Xiaoyan Oral Liquid.

Integrating chemical similarity and bioequivalence: A pilot study on quality consistency evaluation of dispensing granule and traditional decoction of Scutellariae Radix by a totality-of-the-evidence approach

Aims Using both data mining and network pharmacology methods, this paper aims to construct a molecule-target-disease network for medicines used for treating mastitis, mine out targets, and signaling pathways related to mastitis and explore the mechanism of Chinese materia medica (CMM) prescriptions in treating mastitis. Methods A total of 131 CMM prescriptions for treating mastitis were collected from clinical practice and related literatures. A database of prescriptions for treating mastitis (DPTM) was then constructed. Based on data mining method, Traditional Chinese Medicine Inheritance Support System (TCMISS) was employed to mine out high-frequency CMM and key CMM combinations in DPTM. Subsequently, TCM Systems Pharmacology Database and Analysis Platform (TCMSP) and Traditional Chinese Medicine Information Database (TCM-ID) were searched for the targets of ingredients of high-frequency CMM. Then, Bioinformatics Analysis Tool for Molecular Mechanism of TCM (BATMAN-TCM) was searched for diseases and signaling pathways corresponding to the targets of key CMM combinations. The obtained results were denoted as results 1. In addition, human disease database MalaCards was searched for targets and signaling pathways related to mastitis. The obtained results were denoted as results 2. Results 1 and 2 were compared to obtain targets and signaling pathways included in both results, namely, mastitis-related targets of TCMs and mastitis-related signaling pathways that CMM involves in. Then, the biological functions of these targets and signaling pathways were investigated, on which basis the mechanism of CMM prescriptions in treating mastitis was explored. Results A total of 12 key TCM combinations were identified. Taraxaci Herba, Glycyrrhizae Radix et Rhizoma, Paeoniae Radix Alba, semen citri reticulatae, etc. were CMM with the highest frequency of use for treating mastitis. The potential targets of these high-frequency CMM in treating mastitis were intercellular adhesion molecule 1 (ICAM-1), interleukin-6 (IL-6), lipopolysaccharide binding protein (LBP), and lactotransferrin. The potential signaling pathways that key CMM combinations may involve in during mastitis treatment were NF-κB signaling pathway, immune system, PI3K/Akt signaling pathway, and TNF signaling pathway. Conclusions From a perspective of network pharmacology, molecule-target-disease analysis may serve as an entry point for the research of mechanism of CMM. On this basis, we studied the mechanism of CMM prescriptions in treating mastitis by data mining and comparison of results. Our work thus provides a new idea and method for studying the working mechanism of CMM prescriptions.

Gynostemma pentaphyllum (Thunb.) Makino, also known as jiaogulan, has been shown to have antioxidant, antiproliferative, and anti-inflammatory activities. Flavonoid is considered a major contributor for these beneficial effects. To obtain chemical patterns of flavonoids in G. pentaphyllum of different genotypes (di- versus tetraploids) and different parts (leaf versus whole plant) of plants, the extraction condition was optimized and a fingerprinting approach was established by means of high-performance liquid chromatography coupled with diode array detection and mass spectrometry (HPLC-DAD-MS). Eight flavonoids were identified, among which rutin and quercetin were quantified. The highest levels of rutin and quercetin were 23.03 and 12.10 mg/g, respectively, observed in the diploid leaf sample 2L3 and 2L2, while the lowest levels of rutin and quercetin were 1.92 and 0.25 mg/g in the tetraploid whole-plant sample 4W3. The chemical patterns were further analyzed by similarity calculation and principal component analysis (PCA). Seven common characteristic peaks were found in all of the tested samples. Flavonoid patterns of tetraploids were significantly different from those of diploids, whereas different parts of plants showed less difference. The flavonoid pattern of the diploid leaf sample was most similar to that of the reference botanical G. pentaphyllum. The combination of chromatographic fingerprint and quantification analysis could be used for quality assessment of G. pentaphyllum and its derived nutraceutical products.

The effect on the antioxidant activity and Nitric Oxide activity production inhibitory activity of Taraxacum has not been known. Therefore, phenolics and flavonoid contents were investigated from the ethanol extracts of five differ-ent Taraxacum species. The results showed that, among the five Taraxacum, T. hallaisanensis contains the highest total phe-nolic and flavonoid contents. When the antioxidant activity was measured by DPPH, ABTS and reducing power activity, the free radical scavenging activity of T. hallaisanensis was also the highest among five Taraxacum species. However, mea-surement by CCK-8 assay in Raw264.7 cells indicated that the extracts of Taraxacum species have no effect on cell viability. Moreover, we also investigated the effect of Taraxacum species on NO scavenging activity in lipopolysaccharide (LPS)-stim-ulated Raw264.7 cells. The results clearly showed that Taraxacum species inhibited NO production, and the inhibitory effect of T. hallaisanensis was the strongest. The above results suggested that Taraxacum species affected the antioxidant and NO scavenging activity, and among the five species, antioxidant and NO scavenging activity assay of T. hallaisanensis was signif-icantly higher than those of other four Taraxacum species. Therefore, T. hallaisanensis could be used as a potential drug with anti-oxidant and anti-inflammatory effect.

Discovery of discriminatory quality control markers for Chinese herbal medicines and related processed products by combination of chromatographic analysis and chemometrics methods: Radix Scutellariae as a case study

We first investigated liquid chromatography (LC) fingerprint method using multi-wavelength combination technique, and successfully used this method for the analysis of a fat-soluble extract from Radix isatidis. LC fingerprints of fat-soluble R. isatidis extracts from 11 origins were established using the Origin software and Similarity Evaluation System for Chromatographic Fingerprints of traditional Chinese medicine (TCM). The typical LC fingerprints of fat-soluble extracts from R. isatidis were first established, and the reference chromatogram was also generated with 24 common peaks showing large peak areas and good separation from adjacent peaks. Seven common characteristic peaks were identified for the first time: anthranilic acid, syringic acid, benzoic acid, salicylic acid, tryptanthrin, indigo and indirubin. The total peak areas of 24 common peaks were more than 80% of the total peak areas. Hierarchical clustering analysis (HCA) of 11 R. isatidis samples was performed, and the results show that the differences between 11 origin R. isatidis were large. Principal component analysis (PCA) on 24 common peaks was obtained to find the possible chemical markers for the discrimination of different samples. The loading plot indicated that peaks 8, 11, 13 and 14 may have more influence on the discrimination of the samples. All these were useful for evaluating and controlling the quality of R. isatidis. Our work provides a general model of chromatogram combination at multi-wavelength detection to study the complex or the undeveloped materials, which can be used to scientifically ensure the quality of such samples and deeply do qualitative, quantitative and multicomponent pharmacodynamic research combined with modern advanced chromatographic technique.

Biopotency Assays, a Model with Integration Feature for Quality Control Research of CMM

This study summarized and analyzed the current acne-relieving Chinese medicinal health products and Chinese patent medicines,aiming at providing references for the formulation and development of acne-relieving health products. The information on acne-relieving health products published by the Department of Special Food Safety Supervision and Management,the State Administration for Market Regulation( SAMR) was retrieved,and the Chinese patent medicines on DRUGDATAEXPY were searched. Microsoft Excel and the Traditional Chinese Medicine Inheritance Support System V2. 5( TCMISS) were employed to statistically analyze the characteristics of formulations. Forty-three acne-relieving health products were obtained,including 40 ones containing Chinese herbal medicines. Six Chinese herbal medicines showed a usage frequency ≥8,with 61 times in use totally,including Salviae Miltiorrhizae Radix et Rhizoma( 13),Lonicerae Japonicae Flos( 12),Taraxaci Herba( 11),ALOE( 9),Carthami Flos( 8),and Rhei Radix et Rhizoma( 8). A total of 29 Chinese patent medicines with similar efficacy were collected in DRUGDATAEXPY. Nine Chinese herbal medicines showed a usage frequency ≥6,with 63 times in use,including Rhei Radix et Rhizoma( 10),Glycyrrhizae Radix et Rhizoma( 8),Scutellariae Radix( 8),Lonicerae Japonicae Flos( 7),Angelicae Sinensis Radix( 6),Paeoniae Radix Rubra( 6),Astragali Radix( 6),Paeoniae Radix Alba( 6),and Gardeniae Fructus( 6). Through unsupervised hierarchical entropy-based clustering of the above-mentioned Chinese medicinal health products and Chinese patent medicines,five and three new formulas were obtained,respectively. The selection of Chinese herbal medicines was consistent with the principles of traditional Chinese medicine( TCM) theories in relieving acne,i. e.,dispelling exterior wind,ventilating lungs,clearing heat,dissipating mass,promoting diuresis,relaxing bowels,removing toxin,activating blood,resolving stasis,eliminating phlegm,and regulating Qi. According to the " lung governing skin and body hair" and " interior-exterior relationship between lung and large intestine" in TCM theories,the idea that acne could be treated by relieving constipation was proposed,which provided new methods and references for the research and development of Chinese medicinal health products.

Chromatographic fingerprinting technique of traditional Chinese medicine (TCM) has proved to be a comprehensive strategy for assessing the intact quality of herbal medicine. In general, one could use the chromatographic techniques to obtain a relatively complete picture of herbal medicines, which are in common called chromatographic fingerprints of herbal medicines to represent the so-called phytoequivalence. Based on this, the features of chromatographic fingerprints of herbal medicines have been discussed in some detail. The technique based on chromatographic fingerprinting is essentially a kind of high-throughput and integral tools to explore the complexity of herbal medicines. In order to further control the comprehensive quality of TCMs, some new strategies are proposed to trace the chemical changes of chromatographic fingerprints both in product processing and/or after their administration by modern chromatographic techniques and chemometrics. Combined with metabolomics, it seems possible for one to reveal the working mechanism of TCMs and to further control their intrinsic quality. Finally, the intensive study of chromatographic fingerprinting coupled with multivariate analysis tools developed in bioinformatics and chemometrics are emphasized in order to achieve the aim to reveal the working mechanisms of TCMs and to further control and strengthen TCMs' intrinsic quality in a comprehensive manner.

As the biological behaviour of even early stage renal cell cancer (RCC) strongly correlates with tumor size, it has been argued that the inclusion of RCC up to a maximum diameter of 7 cm into a common subgroup classified as T1 according to the 5th edition of the TNM system would not adequately represent the different biological aggressiveness of these malignancies. Taking this into account, the TNM classification, which now categorizes T1 RCC as T1a and T1b according to a cutoff size of 4 cm, was recently modified. However, only a few larger investigations, mainly based on univariate statistical analyses, that support the suitability of this cutoff are at present available from the literature. Therefore, it was the aim of the present investigation to determine the tumor size that best separates patients with low responses from those with high risk for tumor progression by univariate (log rank test) and multivariate (Cox regression model) statistical analyses. Between 1981 and 2000, 652 patients (443 males and 209 females) underwent tumor nephrectomy in our clinic for the diagnosis of RCC. Of these, 243 patients revealed primary tumors with a local growth not extending beyond the renal capsula at the time of surgery. For the different cutoff levels (starting from 2 cm in increments of 1 cm up to 8 cm) that were selected to subdivide the patients into groups according to the maximum tumor diameter, the correlation between tumor size and overall survival was determined by univariate and multivariate statistical analyses. It became evident that although during univariate analysis the prognostic value of a cutoff size of 4 cm was confirmed, multivariate analysis identified the highest relative risk for cause-specific death (2.93) for patients having tumors larger than 5 cm in maximum diameter. Therefore, the 5 cm cutoff seems to best determine the clinical prognosis of patients undergoing tumor nephrectomy for early stage RCC. The present study demonstrates the need for multivariate statistical approaches when the latest modification of the TNM classification system is critically evaluated.