The material basis of bitter constituents in Carbonized Typhae Pollen, based on the integration strategy of constituent analysis, taste sensing system and molecular docking.

The discovery of bitter constituents is of great significance to the exploration of medicinal substances for they have potential physiological effects. Carbonized Typhae Pollen (CTP), which is a typical example of carbonized Traditional Chinese Medicine (TCM), has a bitter taste and hemostatic effect after carbonized processing. The objective of this study is to elucidate the material basis of bitter constituents in CTP. Firstly, the constituents of CTP extracts with 7 different solvents were characterized by UPLC-Q-TOF-MS. Then, multivariate statistical analysis was used to visualize the CTP extracts from 7 solvents. A total of 37 constituents were tentatively identified and 17 constituents were considered as the key constituents in differentiating 7 different solvent extracts. Subsequently, the bitter evaluation of extracts from different polar parts was investigated by using an electronic tongue. As a result, the order of bitterness of the extracts was as follows: ethanol > methanol > water > n-butyl alcohol > petroleum ether > butyl acetate > isopropanol. There were statistically significant differences in the bitter degree of extracts. By correlation analysis of bitter information and chemical constituents with partial least squares regression (PLSR), 8 potential bitterness constituents were discovered, including phenylalanine, valine, chlorogenic acid, isoquercitrin, palmitic acid, citric acid, quercetin-3-O-(2-α-L-rhamnosyl)-rutinoside, and typhaneoside. Additionally, molecular docking analysis was conducted to reveal the interaction of these constituents with the bitter taste receptor. The docking result showed that these constituents could be embedded well into the active pocket of T2R46 and had significant affinity interactions with critical amino acid residues by forming hydrogen bonds. This study provided a reliable theoretical basis for future research on biological activity of bitterness substances.

[Applicability of odor analyzer based on study of processed ginger products and GC-MS validation].

Heracles Ⅱ rapid electronic odor analyzer has been gradually applied in the study on the "odor" of Chinese medicine because of its ability to associate volatile substances in the sample with "odor", but its applicability is not clear. In this article, processed ginger products--dried ginger, roasted ginger, and carbonized ginger were chosen as examples to study the volatile chemical components, and the applied scope and accuracy of Heracles Ⅱ rapid electronic odor analyzer were verified by gas chromatography-mass spectrometry(GC-MS). Components in ten batches of ginger from different producing places were detected by GC-MS and Heracles Ⅱ rapid electronic odor analyzer. Principal component analysis(PCA) and discriminant factor analysis(DFA) were performed to investigate the effects of producing place and processing degree on dried ginger, roasted ginger, and carbonized ginger. The results showed that the processing degree had significant impact on the processed ginger products, while producing place was not significant to the processed ginger products. At the same time, 42 and 45 compounds were detected by Heracles Ⅱ rapid electronic odor analyzer and GC-MS, respectively, of which 29 compounds were common detected, with a coincidence rate of 64.4%. This result indicated that compared with GC-MS, Heracles Ⅱ rapid electronic odor analyzer had certain referential value in the qualitative analysis. Heracles Ⅱ rapid electronic odor analyzer, with the characteristics of convenience, high efficiency, and accuracy in processing data, has certain advantages in the "odor" research aiming to comprehensively evaluate volatile substances, and has a good application prospect in the quality control of Chinese medicine and its processed products.

An effective strategy for distinguishing the processing degree of Polygonum multiflorum based on the analysis of substance and taste by LC-MS, ICP-OES and electronic tongue.

The efficacy of raw and processed products of Polygonum multiflorum (PM) varies greatly. "Nine cycles of steaming and sunning" (NCSS) is recognized as an effective technology in enhancing efficacy and reducing toxicity for PM. In this paper, PM was prepared differently into three groups (including group R, M, and "9"), which represent raw PM, PM processed using the method of Chinese Pharmacopoeia (ChP) and PM processed using traditional NCSS, respectively. The purpose is to establish an effective method to distinguish raw PM from different processed products and highlight the rationality of processing technology. The main organic compounds that could distinguish these three groups of samples were identified by in-depth mining of mass spectral information and various chemometric methods. Level of related metal cations have been quantified and used as another important distinguishing markers. The electronic tongue was utilized to determine the taste traits of aqueous extract from PM. Furthermore, the material basis that caused the difference in taste was discovered according to correlation analysis. In detail, saltiness has the most important contribution associated with the concentrations of K+ and Na+, however, bitterness and astringency were mainly associated with the contents of epicatechin gallate, catechin, procyanidin B1, procyanidin B2 and epicatechin. This study proposed a novel and effective strategy for identification of processing technology of PM. It lays the foundation for clarifying the modern scientific recommendations of processing technology to PM. On the other hand, it also provides a reference for related researches on other traditional Chinese medicine (TCM).

Comparative Investigation for Raw and Processed Products of Euodiae Fructus Based on High-Performance Liquid Chromatography Fingerprints and Chemical Pattern Recognition.

As a traditional Chinese medicine, Euodiae Fructus is widely used due to its analgesic, anti-inflammatory, and antihypertensive effects. However, Euodiae Fructus has also been documented to be toxic, and the toxic effects can be reduced by processing. To distinguish Euodiae Fructus from its processes products and study the changes of raw and processed products before and after processing, we evaluated four auxiliary material processing methods including vinegar, Zingiberis Rhizoma, Coptidis Rhizoma, and Glycyrrhizae Radix et Rhizoma. The raw Euodiae Fructus and four processed Euodiae Fructus samples were analyzed and compared based on the high-performance liquid chromatography (HPLC) fingerprints combined with chemometrics, including principal component analysis (PCA), partial least squares-discriminant analysis (PLS-DA), and principal component analysis-class (PCA-Class). A total of 27 common peaks were obtained by fingerprint analysis. The fingerprint similarity of raw and processed samples was between 0.86-0.999. We also determined the contents of the main active ingredients - Evodiamine and Rutaecarpine. PCA and PLS-DA analyses were used to distinguish between the raw and processed samples of Euodiae Fructus, and 14 chemical markers were screened out. Four kinds of processed products were further analyzed and the results showed that they could be successfully distinguished under the established models, and 12 chemical markers were labeled. PCA-Class results revealed that the classification models constructed in this study had adequate discrimination ability. The method combined with HPLC fingerprinting and multi-component chemical pattern recognition technology could be used to differentiate raw and processed Euodiae Fructus with adequate predictive power. Our findings confirmed the rationality of the pharmacopoeial method and provided a reference for the quality control of the Glycyrrhizae Radix et Rhizoma processed Euodiae Fructus.

The composition differences between small black beans and big black beans from different habitats and its effects on the processing of Polygonum multiflorum.

INTRODUCTION: The roots of Polygonum multiflorum (PM) serve as a classical traditional Chinese medicine (TCM), which has multiple biological activities. However, many cases of hepatotoxicity in PM have been reported in recent years. Processing PM with black beans decoction is one of the typical processing methods to reduce the hepatotoxicity of PM since ancient times. OBJECTIVES: To find potential effective constituents, as well as the optimal variety and origin of black beans for the processing of PM. METHODS: Based on ultrahigh-performance liquid chromatography Q-Orbitrap mass spectrometry (UHPLC-Q-Orbitrap-MS) analysis, we measured the contents of the two potential toxic compounds (emodin-8-O-glucoside and torachrysone-O-hexose) in raw PM (R-PM), PM processed with big black beans (B-PM) and PM processed with small black beans (S-PM). The flow cytometry method analysed the effects of different processed products of PM on apoptosis of L02 cells in different drug concentration. Proton nuclear magnetic resonance (1 H-NMR) and UHPLC-Q-Orbitrap-MS together with multivariate statistical analysis were used to systematically analyse the different components between small black beans (Small-BB) and big black beans (Big-BB) from 30 different habitats. RESULTS: The toxicity was ranked from small to large: S-PM < B-PM < R-PM. Processing PM with black beans could significantly decrease the apoptosis rate of L02 cells, especially when the drug concentration is 80 µg/mL. Besides, we find five differential compounds (α-arabinose, α-galactose, proline, isomer of daidzein and isomer of genistein) may be potential active ingredients. In terms of the black beans collected from 30 producing areas, we find that Small-BB from Weifang in Shandong province was optimum to processing PM, followed by Shangqiu in Henan province, Jilin and Liaoning province. CONCLUSION: The ingredients that affect the processing of PM may be attributed to α-arabinose, α-galactose, proline, isomer of daidzein and isomer of genistein in black beans. When the drug concentration is higher, the effect of reducing the hepatotoxicity of PM is better. Besides, Small-BB was more effective than Big-BB for reducing the toxicity of PM, especially Small-BB from Weifang in Shandong, Shangqiu in Henan province and northeast China.

Comparative investigation of raw and processed products of Gardeniae Fructus and Gardenia jasminoides var. radicans using HPLC coupled with chemometric methods.

As a commonly used traditional Chinese medicine (TCM), Gardeniae Fructus (GF) and its processed products, GF (stir-baked) and GF Praeparatus, have important medicinal value in clinical practice. Gardenia jasminoides var. radicans (GJVR) is a variant of GF, and because of the naming GJVR is often confused in the clinic with GF, resulting in medical misprescriptions. To distinguish GF and GJVR and study the changes before and after processing, the fingerprints of GF and GJVR are presented using HPLC, followed by hierarchical cluster analysis (HCA), principal component analysis (PCA), and partial least squares-discriminant analysis (PLS-DA). GF has purging and choleretic effects, and in this study, we determined the content of main active ingredients to preliminarily assess the GF and GJVR quality from the perspective of material basis. For PCA score plot, the samples fell into six clusters, the cross-validity Q2 (cum) = 0.842 and the cumulative contribution rate R2 x (cum) = 0.988, indicating that the model has a good precision. The results were then corroborated by HCA and PLS-DA method, showing that this methodology can distinguish GF and GJVR and can be used for the comparison of raw and two processed products. According to the model established by PLS-DA, eight components were identified as the most significant variables for discrimination. The results obtained by multiple model methods are consistent and verified by each other, providing a scientific reference for further clarification of the medicinal properties of GF and GJVR.

Quality consistency evaluation of Kudiezi Injection based on multivariate statistical analysis of the multidimensional chromatographic fingerprint.

Traditional Chinese Medicine Injection (TCMI) was restricted due to the batch-to-batch variability caused by the variable compositions of botanical raw materials and complexities of the current manufacturing process. To evaluate and control the quality of Kudiezi Injection (KDZI), a comprehensive and practical method based on multidimensional chromatographic fingerprint associated with multivariate statistical analysis was proposed. The multidimensional chromatographic fingerprint was established by integrating three kinds of chromatographic fingerprints, including High Performance Liquid Chromatography-Ultraviolet spectrum (HPLC-UV), Gas Chromatography-Mass Spectrometer (GC-MS) and High performance ion-exchange chromatography (HPIEC), which were used to detect flavones, nucleosides, organic acids, amino acids and saccharides in KDZI. In addition, four main multivariate statistical analyses were compared to assess the batch-to-batch consistency of samples. Results showed that the cosine method, which has been widely used in the quality evaluation of TCM, failed to distinguish the differences among batches based on neither chromatographic peaks' area nor contents information. t-test and Bayes' theorem could reveal the content difference among batches, while hierarchical clustering analysis could differentiate KDZI batches, and Luteolin-7-O-ß-D-glucuronopyranoside, Tau, Ser, guanine and allose were the main indicators. In conclusion, multidimensional chromatographic fingerprints could reflect the quality information of KDZI comprehensively and hierarchical clustering analysis was suitable to identify the differences among batches. This could provide an integrated method for consistency evaluation of TCMI, process improvement of TCMI and solving similar problems in TCMI.

Establishment of modified biopharmaceutics classification system absorption model for oral Traditional Chinese Medicine (Sanye Tablet).

ETHNOPHARMACOLOGICAL RELEVANCE: As one of the new drugs of traditional Chinese medicine, Sanye Tablet is employed as a hypolipidemic in the traditional medicine, but the biopharmaceutical properties of the drug is still unclear. AIM OF THE STUDY: Through the study of biopharmaceutical properties, the classical biopharmaceutics classification system (BCS) can be used to classify and predict the in vivo absorption properties. On this basis, the biopharmaceutical properties closely related to traditional Chinese medicine preparations are added and a modified BCS model is established to predict and judge the absorption degree of traditional Chinese medicine compound. MATERIALS AND METHODS: Representative components of Sanye Tablet were selected and subjected to different in vitro tests. The experimental results were compared with the results of the BCS to evaluate the accuracy and applicability to Sanye Tablet. We take parameters of dissolution and stability based on product characteristics into account. A "modified-BCS" was developed and the results of the improved method and the classic method were compared. Also the ability of each classification system to predict and determine the extent of absorption of the Chinese herbal compound was investigated based on the absolute bioavailability of representative components. RESULTS: For classic BCS, the five representative components (except for nuciferine) are all class III, nuciferine is class I/II obtained by Caco-2 cell assay and class III/IV obtained by everted gut sac assay. For modified BCS, paeoniflorin is class III, rutin, hyperoside and salvianolic acid B are class III/IV, and nuciferine is class I/II based on Caco-2 cell assay, class III/IV based on everted gut sac assay. Nuciferine is the best of the five components, with absolute bioavailability reaching 61.91% based on in vivo bioavailability test. CONCLUSIONS: The five representative components (except for nuciferine) are all class III/IV, which correlates well with the absolute bioavailability results and demonstrates that they are poorly absorbed substances. The correlation between the classification results obtained using the "modified-BCS" and absorption in the body is better than the correlation obtained using the classic method, suggesting that the improved BCS is more suitable for the characterization of Sanye Tablet. These results indicate that the oral formulation of Sanye Tablet is a BCS III/IV drug.

Rapid Discovery of the Potential Toxic Compounds in Polygonum multiflorum by UHPLC/Q-Orbitrap-MS-Based Metabolomics and Correlation Analysis.

The dry roots of Polygonum multiflorum (PM), involving both the raw and processed materials, are widely used as the traditional Chinese medicine for treating various diseases in China. Hepatotoxicity has been occasionally reported in patients who consume PM. Unfortunately, no definite criteria are currently available regarding the processing technology of PM for reduction the toxicity. In this work, we aimed to investigate the variations of PM metabolite profiles induced by different processing technologies by UHPLC/Q-Orbitrap-MS and multivariate statistical analysis, and to discover the potential toxic compounds by correlating the cytotoxicity of L02 cell with the contents of metabolites in raw and processed PM samples. We could identify two potential toxic compounds, emodin-8-O-glucoside and torachrysone-O-hexose, which could be selected as the toxic markers to evaluate different processing methods. The results indicated all processed PM samples could decrease the cytotoxicity on L02 cell. The best processing technology for PM process was to steam PM in black soybean decoction (BD-PM) for 24 h.

Comparison of the Active Compositions between Raw and Processed Epimedium from Different Species.

Epimedium herb is one of the most vital traditional Chinese medicines (TCMs), which is used for "nourishing the kidney and reinforcing the Yang". In the guidance of TCM theory, Epimedium herb is usually processed with lamb oil to increase its efficacy. The contents of active ingredients in different Epimedium are significantly varied, which may derive from their different species, regions and processing methods. In this research, 13 batches of raw Epimedium collected from 6 provinces were identified. After optimization of the processing method of Epimedium, a liquid chromatography⁻mass spectrometry (LC⁻MS/MS) method for simultaneous determination of 16 compounds was established to evaluate the quality of raw and processed. Then the multivariate statistical technique was applied to compare different batches of Epimedium based on the LC⁻MS/MS data. As a conclusion, the herbs collected from 6 areas were ascribed to 5 species by microscopic and appearance features. Meanwhile, all of the raw and processed samples were classified by partial least squares discriminant analysis (PLS-DA) based on the 16 analyzed compounds. The comparison results indicate that processing and species both have important influences on Epimedium compositions contents.

Effect of wine and vinegar processing of Rhizoma Corydalis on the tissue distribution of tetrahydropalmatine, protopine and dehydrocorydaline in rats.

Vinegar and wine processing of medicinal plants are two traditional pharmaceutical techniques which have been used for thousands of years in China. Tetrahydropalmatine (THP), dehydrocorydaline (DHC) and protopine are three major bioactive molecules in Rhizoma Corydalis. In this study, a simple and reliable HPLC method was developed for simultaneous analysis of THP, DHC and protopine in rat tissues after gastric gavage administration of Rhizoma Corydalis. The validated HPLC method was successfully applied to investigate the effect of wine and vinegar processing on the compounds' distribution in rat tissues. Our results showed that processing mainly affect the T(max) and mean residence time (MRT) of the molecules without changing their C(max) and AUC(0-24)( )(h) Vinegar processing significantly increased the T(max) of DHC in heart, kidney, cerebrum, cerebrellum, brain stem and striatum and prolonged the T(max) of protopine in brain. No significant changes were observed on the T(max) of THP in rat tissues after vinegar processing. Wine processing reduced the T(max) of protopine and DHC in liver and spleen and T(max) of protopine in lung, but increased the T(max) of THP in all the rat tissues examined. To our knowledge, this is the first report on the effects of processing on the tissue distribution of the bioactive molecules from Rhizoma Corydalis.

[Comparison of pharmacokinetics of tetrahydropalmatine monomer and extractive of corydalis and corydalis processed with vinegar].

OBJECTIVE: To investigate the pharmacokinetics of tetrahydropalmatine monomer and extractive of corydalis and corydalis processed with vinegar in rats. METHODS: The plasma concentrations of tetrahydropalmatine were determined by a reversed phase high performance liquid chromatographic. The plasma concentrations-time data were calculated with 3p97 program. RESULTS: The main pharmacokinetic parameters of tetrahydropalmatine monomer and extractive of corydalis and corydalis processesd with vinegar as follows: T(1/2) was (5.66 +/- 1.92), (4.24 +/- l1.54), (4.35 +/- 1.34) h, Tmax was (1.5 +/- 0.5), (1.0 +/- 0.55), (0.5 +/- 0.68) h, Cmax was (0.71 +/- 0.29), (0.37 +/- 0.11), (0.67 +/- 0.35) microg/ml,AUC(o-t), was (2.58 +/- 0.85), (1.96 +/- 0.69), (2.95 +/- 1.61) microg/ml. CONCLUSION: The T(1/2) of corydalis and corydalis processesd with vinegar is more shorter than tetrahydropalmatine monome, the effect of preventing pain is promoted in rats.