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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.

Unsaturated fatty acids (UFAs) are fatty acids with one or more carbon-carbon double bonds (C=C). They are building blocks of structurally complex lipids. According to the position of C=C bond in the UFAs, there are multiple isomers with different physiological activities. Therefore, an accurate understanding of UFAs is essential. Combining Paternò-Büchi (PB) reaction with tandem mass spectrometry (MS/MS) analysis has been successfully used to identify unsaturated lipids in bio-samples. However, there are still some challenges, such as low reactive rate and low MS response of PB products. In this study, 3-pyridinecarboxaldehyde (3-PYA) was selected as an efficient PB reagent for the first time. According to the nitrogen rule, introduction of nitrogen did not only make the target MS and MS/MS ions to be easily identified but also increase the ionization efficiency significantly. The developed method in this study demonstrated greater efficiency and sensitivity for the identification of UFAs in total lipid extracts from bovine liver compared with other commonly used PB reagents.

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.

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.

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).

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-β-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.

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.

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Epimedium herb is one of the most vital traditional Chinese medicines (TCMs), which is used for &ldquo, nourishing the kidney and reinforcing the Yang&rdquo, 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&ndash, mass spectrometry (LC&ndash, 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&ndash, 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.

To analyze the effect of crosswind on the handling stability of tractor semi-trailers in double-lane change situations, the tractor semi-trailer model and the relative aerodynamic coefficient model were built by using TruckSim. Considering the factors of different crosswind directions, crosswind speed, and vehicle speed, we analyzed the relationship between these factors and the handling stability of tractor semi-trailers in double-lane change situations. The results show that a trailer has movement delay and worse handling stability compared to a tractor. The tractor/semi-trailer is unstable when the vehicle speed is 75 km/h and the crosswind speed is 40km/h or 60km/h. The tractor semi-trailer is stable when the vehicle speed is 70 km/h and the crosswind speed is less than 60km/h.

Capillary electrophoresis (CE) with the use of mass spectrometry (MS) has been considered as a unique tool for microscale enzyme assay and inhibitor screening. In this study, matrix metalloproteinase-9 (MMP-9) was selected as target enzyme due to its important role in tumor invasion and metastasis. In order to define the optimal MS parameters, a two level half fraction factorial experimental design was performed. A background electrolyte consisting of 20mM ammonium acetate (pH 6.8) and a sheath liquid of water-methanol (50:50, v/v) containing 0.05% formic acid at a flow rate of 4μl/min were selected. This system was operated in the positive ion mode with a detection-limit of 10nM for the MMP reaction product and provided 60 folds enhancement of sensitivity by using selected reaction monitoring detection compared with MS full scan mode, which significantly increased the detectability of the system and therefore reduced the enzyme reaction time in both off-line and in-line mode. Both electrophoretically mediated microanalysis and pressure mediated microanalysis combined with MS detection were investigated for MMP inhibitor screening. Good repeatability (RSD of peak area and migration time were lower than 5.0%) and linearity (R(2)>0.996) were obtained for both in-capillary approaches. Several tetracycline antibiotics and natural products were selected to test the system. The results indicated an agreement on the ranking of inhibitory potency for both in-capillary approaches.