Identification of Dalbergiae Odoriferae Lignum and Its Counterfeits by 1H-NMR Combined with Multivariate Statistical Analysis / 中国实验方剂学杂志

ObjectiveTo establish the identification method of Dalbergiae Odoriferae Lignum（DOL） and its counterfeits by nuclear magnetic resonance hydrogen spectrum（1H-NMR） combined with multivariate statistical analysis. Method1H-NMR spectra of DOL and its counterfeits were obtained by NMR， and the full composition information was established and transformed into a data matrix， and the detection conditions were as follows：taking dimethyl sulfoxide-d6（DMSO-d6） containing 0.03% tetramethylsilane（TMS） as the solvent， the constant temperature at 298 K（1 K=-272.15 ℃）， pulse interval of 1.00 s， spectrum width of 12 019.23 Hz， the scanning number of 16 times， and the sampling time of 1.08 s. Similarity examination and hierarchical cluster analysis（HCA） were performed on the data matrix of DOL and its counterfeits， and orthogonal partial least squares-discriminant analysis（OPLS-DA） was used to analyze the data matrix and identify the differential components between them. In the established OPLS-DA category variable value model， the category variable value of DOL was set as 1， and the category variable value of the counterfeits was set as 0， and the threshold was set as ±0.3， in order to identify the commercially available DOL. The OPLS-DA score plot was used to determine the types of counterfeits in commercially available DOL， and it was verified by thin layer chromatography（TLC）. ResultThe results of similarity analysis and HCA showed that there was a significant difference between DOL and its counterfeits. OPLS-DA found that the differential component between DOL and its counterfeits was trans-nerolidol. The established category variable value model could successfully identify the authenticity of the commercially available DOL. The results of the OPLS-DA score plot showed that there were heartwood of Dalbergia pinnata and D. cochinchinensis in the commercially available DOL， and were consistent with the TLC verification results. ConclusionThere is a phenomenon that heartwood of D. pinnata and D. cochinchinensis are sold as DOL in the market. 1H-NMR combined with multivariate statistical analysis can effectively distinguish DOL and its counterfeits， which can provide a reference for the identification of them.

Identification of the New Psychoactive Substance Eutylone / 法医学杂志

OBJECTIVES@#To establish a method to identify unknown sample based on the combined use of Fourier transform infrared spectroscopy (FTIR), gas chromatography-quadrupole time-of-flight mass spectrometry (GC-QTOF-MS), ultra-high performance liquid chromatography-linear ion trap quadrupole-orbitrap mass spectrometry (UPLC-LTQ-Orbitrap MS) and 1H-nuclear magnetic resonance spectroscopy (1H-NMR) technique.@*METHODS@#The unknown sample was directly analyzed by FTIR. The unknown sample was dissolved in methanol solution containing internal standard SKF525A and the supernatant was detected by GC-QTOF-MS and UPLC-LTQ-Orbitrap MS. The unknown sample was dissolved in methanol-d4 solution for structural analysis of 1H-NMR.@*RESULTS@#The characteristic absorption peaks of FTIR spectra obtained from unknown sample were 1 682 (C=O bond), 1 503, 1 488, 1 436, 1 363, 1 256, 1 092, 1 035, 935, 840 and 800 cm-1, the characteristic fragment ions (m/z) of GC-QTOF-MS were 86.096 4 (base peak), 58.065 1, 149.023 5, 121.028 6 and 65.038 6, the accurate mass [M+H]+ detected by UPLC-LTQ-Orbitrap MS was 236.127 7. The sample was identified as synthetic cathinone new psychoactive substance Eutylone by 1H-NMR.@*CONCLUSIONS@#The method established in this study can be used for structural confirmation of Eutylone.

Characterization of multiple chemical components of GuiLingJi by UHPLC-MS and 1H NMR analysis / 药物分析学报

GuiLingJi(GLJ),a classic traditional Chinese medicine(TCM)formula,is composed of over 20 herbs,according to the Pharmacopeia of the People's Republic of China.Owing to its various activities,GLJ has been used in clinical settings for more than 400 years in China.However,the ambiguous chemical material basis limits the development of studies on the quality control and pharmacological mechanisms of GLJ.Therefore,comprehensive characterization of the multiple chemical components of GLJ is of great significance for the modernization of this formula.Given the great variety of herbs in GLJ,both UHPLC-MS and 1H NMR techniques were employed in this study.In addition,solvent extraction with different polarities was used to eliminate signal interference and the concentration of trace components.A variety of MS analytic methods were also used,including implementation of a self-built compound database,diagnostic ion filtering,mass defect filtering,and Compound Discoverer 3.0 analysis software.Based on the above strategies,a total of 150 compounds were identified,including 5 amino acids,13 phenolic acids and glycosides,11 coumarins,72 flavones,20 triterpenoid and triterpenoid saponins,23 fatty acids,and 6 other compounds.Moreover,13 compounds were identified by 1H NMR spectroscopy.The UHPLC-MS and 1H NMR results supported and complemented each other.This strategy provides a rapid approach to analyzing and identifying the chemical composition of Chinese herbal prescriptions.The current study provides basis for further research on the quality control and pharmacological mechanism of GLJ.

Fingerprint of Shenmai Injection based on ~1H-NMR technique / 中国中药杂志

Shenmai Injection is a Chinese medicinal injection prepared from Ginseng Radix et Rhizoma Rubra and Ophiopogonis Radix, which is widely used in clinical practice for the treatment and adjuvant therapy of cardiovascular diseases with significant pharmacological effects. Proton nuclear magnetic resonance spectroscopy(~1H-NMR) has the advantages of simple and nondestructive sample pretreatment, fast analysis, abundant chemical information, quantification and no need to follow the standard curve. It is widely used in the analysis and research of complex mixtures of traditional Chinese medicine, clinical blood and urine samples. In this study, the ~1H-NMR fingerprint of Shenmai Injection was established. Thirty-two chemical components were identified, including seven amino acids, eight small molecular organic acids, one alkaloid, four sugars, two nucleosides, seven saponins, and three other components. Pearson's correlation coefficient and multivariate analysis of variance(principal component analysis combined with hierarchical cluster analysis) were applied based on the ~1H-NMR fingerprint to evaluate the quality consistency. The results showed high-quality consistency of 82 batches of Shenmai Injection. This study confirms that the ~1H-NMR fingerprint has great potential in the application of quality control of Chinese medicinal injection.

In vitro pharmacological attributes and metabolite's fingerprinting of Conocarpus lancifolius / Atributos farmacológicos in vitro y huellas dactilares de metabolitos de Conocarpus lancifolius

Search for safe antioxidants and novel nutraceuticals urged to evaluate the antioxidant, anti-acetylcholine esterase and anti-lipoxygenase activity of various leaf extracts of Conocarpus lancifolius. Extraction was optimized from freeze dried plant extracts quenched with liquid nitrogen using water, ethanol, methanol, hexane, ethyl acetate and chloroform. Maximum extract yield, total phenolic contents and total flavonoid contents were obtained in case of ethanolic extraction. The highest 2,2-diphenyl-1-picrylhydrazylradical scavenging in terms of IC50 value of 55.26 µg/mL was observed for ethanolic leaf extract. The acetylcholine esterase and lipoxygenase inhibitory activities (IC50) were also observed for ethanolic extract. These findings for ethanolic extract were statistically significant when compared with other extracts (ρ<0.05). The haemolytic % values indicated that all extracts were associated with very low or negligible toxicity. The epicatechin, isorhamnetin, rutin, scopoleptin, skimmianine, quercetin-3-O-α-rhamnoside, quercetin-3-O-ß-glucoside, cornoside, creatinine, choline, pyruvic acid, α-hydroxybutyric acid, phyllanthin and hypophyllanthin were identified as major functional metabolites in ethanolic leaf extract of C. lancifoliusby 1H-NMR. The identified metabolites were probably responsible for the pharmacological properties of C.lancifolius. The findings may be utilized as pharmacological leads for drug development and food fortification.

Se insta a la búsqueda de antioxidantes seguros y nuevos nutracéuticos para evaluar la actividad antioxidante, anti-acetilcolina esterasa y anti-lipoxigenasa de varios extractos de hojas de Conocarpus lancifolius. La extracción se optimizó a partir de extractos de plantas liofilizados enfriados con nitrógeno líquido usando agua, etanol, metanol, hexano, acetato de etilo y cloroformo. En el caso de extracción etanólica se obtuvo el rendimiento máximo de extracto, el contenido de fenoles totales y el contenido de flavonoides totales. La mayor eliminación de radicales 2,2-difenil-1-picrilhidrazilo en términos de valor de CI50 de 55,26 µg/mL se observó para el extracto de hoja etanólico. También se observaron las actividades inhibidoras de la acetilcolina esterasa y lipoxigenasa (CI50) para el extracto etanólico. Estos hallazgos para el extracto etanólico fueron estadísticamente significativos en comparación con otros extractos (ρ<0.05). Los valores del % hemolítico indicaron que todos los extractos estaban asociados con una toxicidad muy baja o insignificante. Se identificaron la epicatequina, isorhamnetina, rutina, escopoleptina, skimmianina, quercetina-3-O-α-ramnosido, quercetina-3-O-ß-glucósido, cornosido, creatinina, colina, ácido pirúvico, ácido α-hidroxibutírico, filantrina e hipofillantina. como metabolitos funcionales principales en el extracto etanólico de hojas de C. lancifoliuspor 1H-NMR. Los metabolitos identificados probablemente fueron responsables de las propiedades farmacológicas de C. lancifolius. Los hallazgos pueden utilizarse como pistas farmacológicas para el desarrollo de fármacos y la fortificación de alimentos.

Bile acid derivatives-focused chemical profiling in snake bile / 中国中药杂志

As a precious traditional Chinese medicine(TCM), snake bile has been widely used in numerous Chinese medicine prescriptions. Bile acid(BA) derivatives have been demonstrated as the primary chemical family in snake bile. In-depth chemical characterization of BAs is of great importance towards the establishment of quality standards and clarification of the effective material basis for snake bile. This study firstly employed ~1H-NMR to preliminarily analyze the chemical profiles of snake bile, an automated fraction collector was subsequently implemented to obtain the fractions-of-interest. The fraction was then concentrated and re-analyzed by LC-MS. Based on ~1H-NMR, BAs were found to be the main components of snake bile, and six BAs including CDCA, CA, TCDCA, TCA, TDCA and GCA were tentatively identified from the representative spectrum with the assistance of literature and reference compounds. Whereas the content of TCA in snake bile was too great, resulting in a great obstacle for the detection of trace components, the automated fraction collector was subsequently implemented to obtain the fractions-of-interest for LC-MS analysis. According to matching MS/MS information and retention time with reference compounds as well as database retrieval, a total of 57 BAs were detected and annotated. Because of the combination of ~1H-NMR and LC-MS platforms, the findings are beneficial for the in-depth characterization of BAs in snake bile, which provides references for the establishment of quality control and evaluation methods of snake bile.

Study on mechanism of enhancing antioxidant activity in fermented Ziziphi Spinosae Semen with Poria cocos / 中国中药杂志

In this study, the antioxidant property changes in fermented Ziziphi Spinosae Semen(FZSS) with Poria cocos were analyzed by DPPH, ABTS and FRAP methods. Then the content determination of active ingredients and ~1H nuclear magnetic resonance(~1H-NMR) spectroscopy were also used to investigate the mechanism of FZSS with P. cocos in enhancing the antioxidant activity. The results showed that the content of active ingredients such as total phenols, total saponins and total polysaccharides were significantly increased during the fermentation time. The results of ~1H-NMR metabonomics showed that the contents of amino acids such as leucine, lysine, valine and alanine, nitrogen compounds such as creatine, creatinine, and betaine, and secondary metabolites, for instance, jujuboside A and spinosin were higher after fermentation, and above components showed positive correlation with antioxidant capacity in Pearson correlation analysis. Therefore, it was inferred that the enhancement of antioxidant activity of FZSS may be the result of the joint action of various chemical components. This study preliminarily clarified the mechanism of FZSS in enhancing the antioxidant activity, and provided new research ideas for the product development and utilization of FZSS.

In vitro impact of condensed tannins on the caecal metabolites of chickens / Impacto in vitro de taninos condensados nos metabólitos cecais de frangos de corte

Tannins are a diverse group of plant phenolic compounds. Condensed tannins (CTs) represent a major subgroup of tannins and were extracted from tilia (Tilia L.) flowers and black locust (Robinia pseudoacacia) leaves. These extracts were examined for their effects on the metabolic profile of chicken caeca. By using in vitro, a nuclear magnetic resonance (1H-NMR), which was combined with multivariate statistics, the current study was applied for the first time to investigate how three different CT compositions, procyanidins (PC) and/or prodelphinidins (PD) units influenced the metabolic end-products in caecal contents of chickens. In the presence of tannins, glutamate, leucine, lysine, pyroglutamate, phenylalanine, proline, and sarcosine were significantly decreased. CT extracts significantly influenced the fermentation, increasing the concentrations of some fatty acids such as acetate, butyrate, and propionate whereas. In contrast, lactate decreased between the treatments. This study identified the key structural features of CTs that contain either high molar proportions of PD or PC, which might be useful to improve the efficiency of feed utilization in chickens.(AU)

Taninos são um grupo diversificado de compostos fenólicos derivados de plantas. Os taninos condensados (TCs) representam o maior subgrupo de taninos extraídos das flores de tília (Tilia L) e de folhas negras (acácia-bastarda). Estes extratos foram examinados para a avaliação dos seus efeitos no perfil metabólico do ceco de frangos de corte. Com o emprego da ressonância magnética nuclear in vitro (1H-NMR) combinada com estatística multivariada, o presente trabalho foi aplicado pela primeira vez para investigar como três diferentes composições de TCs, unidades de procianidinas (PC) e/ou prodelfinidinas (PD) influenciariam os produtos metabólicos finais dos conteúdos cecais de frangos de corte. Na presença de taninos, houve um significativo decréscimo de glutamato, leucina, lisina, piroglutamato, fenilalanina, prolina e sarcosina. Os extratos de TCs influenciaram significativamente a fermentação, aumentando as concentrações de alguns ácidos graxos, tais como o acetato, butirato e propionato, enquanto em contraste, houve um decréscimo do lactato entre os tratamentos. Este trabalho identificou aspectos estruturais chave que os TCs contêm, tanto as altas proporções molares de PD como as de PC, as quais podem ser úteis para aumentar a utilização de alimentos em frangos de corte.(AU)

Anti-depression mechanisms of Xiaoyao Powder based on fecal metabolomics / 中草药

Objective: To characterize the endogenous metabolites and metabolic changes of feces of chronic unpredictable mild stress (CUMS) rats by 1H-NMR, evaluate the improvement effects of Xiaoyao Powder and investigate the underlying mechanisms. Methods: The depression model was established by CUMS procedure. 1H-NMR coupled with multivariate statistical analysis was employed to reveal the changes of fecal metabolic profiles of CUMS rats and identify potential bio-markers involved in CUMS-induced depression. Based on the potential bio-markers, the relevant metabolic pathways were constructed. Results: A total of 10 metabolites was identified as potential bio-markers in fecal samples for the CUMS model. Compared with the control group, the contents of asparagine, aspartate, lactate and propionic acid in the CUMS rats were significantly increased (P < 0.05, 0.01), while phenylalanine, tyrosine, glutamate, glutamine, alanine and proline were significantly decreased (P < 0.05, 0.01). The administration of Xiaoyao Powder could significantly increase the levels of phenylalanine, tyrosine, glutamate, glutamine and proline, whereas reduced the levels of asparagine, lactate and propionic acid. Compared with the control group, six metabolic pathways were recognized as the most influenced pathways associated with the CUMS-induced depression: (1) aminoacyl-tRNA biosynthesis, (2) alanine, aspartate and glutamate metabolism, (3) arginine and proline metabolism, (4) glutamine and glutamate metabolism, (5) phenylalanine metabolism and (6) pyruvate metabolism. Among them, Xiaoyao Powder significantly mediated abnormalities of five pathways of (2), (3), (4), (5) and (6). Conclusion: It is the first report to investigate the antidepressant-like effects and underlying mechanisms of Xiaoyao Powder from the perspective of fecal metabolites. The current results showed that the anti-depression mechanisms of Xiaoyao Powder might be related to regulating the amino acid metabolism, glucose metabolism and intestinal microbial metabolism. This study provides a solid basis for revealing the anti-depression mechanisms of Xiaoyao Powder comprehensively and deeply.

Effects of Renshenjian Decoction on blood and urine metabonomics in type 2 diabetes rats based on ~1H-NMR / 中国中药杂志

Proton nuclear magnetic resonance(~1H-NMR) is used to investigate the effect of Renshenjian Decoction on serum and urine metabolism of type 2 diabetic rats with insulin resistance induced by high-sugar and high-fat diet combined with low-dose streptozotocin(STZ). After the successful establishment of the insulin resistance model of type 2 diabetes, administration for 35 days, the serum and urine of rats were taken. Once the ~1H-NMR data have been collected and processed, PCA and OPLS-DA were used to analyze them. The results show that: compared with the blank group, the contents of methionine, taurine, α-glucose and β-glucose in the serum of the model group increased significantly(P<0.001), while the contents of 3-hydroxybutyric acid, lactic acid and unsaturated fatty acids decreased significantly(P<0.01). In the model group, the contents of trimethylamine oxide, glycine, α-glucose, β-glucose, taurine and phosphocholine in urine increased significantly(P<0.05), while the contents of creatine, lactic acid, acetic acid and citric acid decreased significantly(P<0.05). Compared with the model group, the contents of 3-hydroxybutyric acid and unsaturated fatty acids in serum of rats in the treatment group increased significantly(P<0.05), while the contents of taurine, α-glucose and β-glucose decreased significantly(P<0.01). In the treatment group, the contents of lactic acid, taurine and creatine in urine increased significantly(P<0.05), while the contents of trimethylamine oxide, glycine, α-glucose, β-glucose and phosphocholine decreased significantly(P<0.01). The results show that Renshenjian Decoction can regulate metabolic disorder and promote the metabolic phenotype to return to the normal range. It displayed therapeutic effect on type 2 diabetic rats with insulin resistance and provided a certain scientific basis for the biological basic research of Renshenjian Decoction by improving insulin resistance in diabetes mellitus.

Analysis on Chemical Characteristics of Lycii Fructus in Qinghai Province Based on 1H-NMR Plant Metabolomics / 中国实验方剂学杂志

Objective:The chemical differences of Lycii Fructus samples from Qinghai, Ningxia, Gansu, Xinjiang and Inner Mongolia provinces were compared based on proton nuclear magnetic resonance (1H-NMR) plant metabolomics. Method:A total of 97 Lycii Fructus samples from five provinces were collected, including 61 samples in Qinghai, and extracted by 50% methanol for detecting. 1H-NMR spectra were obtained and compared by multivariate statistical analysis for investigating the chemical differences of samples from Qinghai and other production areas. And the content of Lycii Fructus polysaccharides in all samples was determined with the wavelength of 490 nm (calculated by anhydrous glucose). Result:A total of 32 chemical components were detected in the Lycii Fructus extract by 1H-NMR. The multivariate statistical analysis revealed that there was no significant difference among the samples from five provinces. The difference between Lycii Fructus from Qinghai and Ningxia, as well as the samples among the six regions of Qinghai province were relatively small. The similarity values of the majority of samples were >0.85. Univariate analysis showed that no significant difference was observed for the most metabolites in Lycii Fructus collected from five provinces, except for sucrose, glucose, proline and so on. There was no significant difference in the content of Lycii Fructus polysaccharides between Qinghai and other provinces. And the correlation coefficient between the content of Lycii Fructus polysaccharides and the small molecular compounds identified by 1H-NMR was -0.2-0.4. Conclusion:In this study, chemical characteristics of Lycii Fructus in Qinghai province are analyzed from the holistic view by 1H-NMR plant metabolomics, in combination of polysaccharide determination, and the results show that there is no significant difference between samples from Qinghai and other four provinces. The quality evaluation method based on 1H-NMR established in this study can provide scientific basis for improving quality control level and selecting planting areas of Lycii Fructus.

Mechanism Analysis of Lung Injury Induced by Water Elution Section of Siegesbeckiae Herba on Rats by 1H-NMR Metabolomics / 中国实验方剂学杂志

Objective:Metabolomics was used to analyze the dynamic changes of endogenous metabolites in serum and lung tissue of rats treated with water elution section of Siegesbeckiae Herba (SWES), and to explore the possible mechanism of lung injury and to search for the sensitive markers in serum and lung tissue. Method:SD rats were randomly divided into three groups, namely the normal group (normal saline), SWES high-dose group and SWES low-dose group (0.500, 0.125 g·kg-1·d-1). SWES was given for 4 weeks and stopped for 2 weeks. The weight and pathological examination were regarded as observation parameters. Serum and lung tissue samples were detected by nuclear magnetic resonance hydrogen spectrum (1H-NMR). The metabolites in rats were analyzed by partial least squares-discriminant analysis (PLS-DA) and orthogonal partial least squares-discriminant analysis (OPLS-DA). Result:Lung inflammation was shown in SWES high-dose group and returned to normal after withdrawal for 2 weeks. The metabolic spectrum of SWES high-dose group was significantly different from the normal group. There were 11 metabolites were identified by 1H-NMR metabolomics, four differential metabolites were identified in serum [acetate, trimethylamine oxide (TMAO), glycine, myo-inositol] and nine differential metabolites were identified in lung tissue [lactate, acetate, phosphatidylcholine (PC), pyruvate, dimethylamine, glutamate, glycerophosphocholine (GPC), glycine, xanthine]. Lung injury was related to the disorder of pyruvate metabolism, glutamate metabolism and other pathways. Most of the metabolites in lung tissue had obviously levels of callback after drug withdrawal, which coincided with the pathological examination. Conclusion:The lung is one of the damaged organs caused by SWES, and the lung injury is reversible and may be related to energy metabolism and oxidative stress.

1H-NMR Metabolomic Analysis of Brain Tissue of CUMS-Induced Depressed Mice Intervened by Iridoid Part of Valerianae Jatamansi Rhizoma et Radix / 中国实验方剂学杂志

Objective:Metabolomics was used to analyze the brain tissue samples of model mice with chronic unpredictable mild stress (CUMS) depression, in order to find out the differential metabolites related to depression and to explore the possible antidepressant mechanism of iridoid part of Valerianae Jatamansi Rhizoma et Radix (IEFV). Method:Forty-two Kunming mice were randomly divided into 6 groups, including the normal group, the model group, the fluoxetine group (2.5 mg·kg-1) and the IEFV low, medium, and high dose groups (doses were 5.73, 11.47, 22.94 mg·kg-1, respectively). The behavioral and biochemical indicators of CUMS model mice were used for pharmacodynamic evaluation with IEFV and a positive drug (fluoxetine) as the intervention drugs. Then, the effect of IEFV on endogenous substances of the brain tissue in CUMS model mice were analyzed by nuclear magnetic resonance hydrogen spectrum (1H-NMR) metabolomics, and multivariate statistical analysis was used to identify the differential metabolites and to enrich the metabolic pathways involved in the differential metabolites. Result:After modeling, the immobility time of the model mice increased significantly, their sucrose preference rate and the excitatory neurotransmitters [5-hydroxytryptamine (5-HT) and norepinephrine (NE)] decreased significantly, indicating the success of modeling. The depression was relieved after IEFV administration, mainly manifested by the recovery of the immobility time, sucrose preference rate and the excitatory neurotransmitters (5-HT and NE). Principal component analysis (PCA) of endogenous metabolites in brain tissue showed that the model group could be significantly separated from the normal group, while the IEFV groups and fluoxetine group all showed a trend of deviating from the model group to the normal group, which was consistent with the behavioral results. The results of orthogonal partial least squares discriminant analysis (OPLS-DA) showed that there were 16 different metabolites between the model group and the normal group, including 12 water-soluble metabolites and 4 liposoluble metabolites. Seven potential metabolism pathways were obtained through MetPA analysis, including metabolism of phenylalanine, metabolism of phenylalanine, tyrosine and tryptophan, metabolism of taurine and hypotaurine acid, metabolism of alanine, aspartic acid and glutamic acid, biosynthesis of valine, leucine and isoleucine, metabolism of D-glutamine and D-glutamate and tricarboxylic acid cycle (TCA). IEFV-high dose group could significantly recall 11 differential metabolites. Conclusion:IEFV may play an antidepressant role mainly by affecting energy metabolism, amino acid metabolism and neurotransmitter levels, which provides a reference for further study on the antidepressant mechanism of IEFV.

Comparison of Chemical Constituents Differences between Angelicae dahuricae with and without Sulphur- fumigation and Selection of Chemical Markers Based on 1H-NMR Metabolomics / 中国药房

OBJECTIVE：To study the diff erences in the overall chemical constituents of Angelicae dahuricae with and without sulphur-fumigation，and to select chemical markers that can be used for identification. METHODS ：Each 16 batches of A. dahuricae with and without sulphur-fumigation were selected as samples. Deuterated methanol-deuterium oxide （1 ∶ 0.2，V/V）was used as extraction solvent ，and 1H-NMR metabolomics was used to detect the primary and secondary metabolites in A. dahuricae . In addition ，visual analysis ，principal component analysis （PCA）and t-test were used to compare the component differences of A. dahuricae by two kinds of pretreatment methods. RESULTS ：A total of 19 chemical constituents such as coumarins ，amino acids and sugars were simultaneously detected and identified from two kinds of A. dahuricae . Visual analysis showed that the chemical profiles of the two kinds of A. dahuricae were similar ，but their coumarins and carbohydrates were quite different in 1H-NMR signal intensity. PCA analysis showed that all samples could be divided into two categories according to different treatment methods. The results of t-test showed that the contents of oxypeucedanin ，imperatorin，glucose and sucrose of A. dahuricae were decreased significantly after sulphur fumigation （P＜0.01 or P＜0.001），while the contents of alanine and leucine were significantly increased （P＜0.01 or P＜0.001）. There was no statistical significance in the contents of other 13 kinds of components （P＞0.05）. CONCLUSIONS：There are obvious differences in the 6 chemical compounds of oxypeucedanin ，imperatorin，glucose，sucrose， alanine and leucine in A. dahuricae with and without sulphur-fumigation ，which can be used as chemical markers for the identification of sulphur-fumigated A. dahuricae in the market.

Study on Antitumor Mechanism of Periplaneta americana Extract C Ⅱ-3 on MFC Tumor-bearing Mice Based on 1H-NMR Metabonomics / 中国药房

OBJECTIVE：To preliminarily study the antitumor mechanism of Periplaneta americana extract C Ⅱ-3 on MFC tumor-bearing mice. METHODS ：Balb/c mice were randomly divided into model group （normal saline 20 mL/kg）and C Ⅱ-3 group （200 mg/kg），with 6 mice in each group. MFC cell suspension （0.2 mL）was injected under the right armpit of mice. On the next day，mice were given relevant medicine intragastrically ，once a day ，for consecutive 10 d. 24 h after the last administration ，Based on the measurement of tumor size ， 1H-NMR technology combined with unsupervised PCA ，supervised PLS-DA and OPLS-DA were used to compare metabolic spectrum of liver tissue from tumor-bearing mice of 2 groups，to analyze differential metabolites and to explore the potential antitumor mechanis m of C Ⅱ -3. RESULTS ：Compared with model group ，the tumor body was significantly reduced in tumor-bearing mice of C Ⅱ-3 group. There were differences in 1H-NMR spectra between the 2 No.81960712）； groups. According to unsupervised PCA ，supervised PLS-DA and OPLS-DA ，totally six potential differential metabolites ，as glycogen （increased），pyruvate （decreased），arginine （de- creased），hydroxyproline （increased），inosine （increased） and niacinamide （increased），were identified in the liver tissue，which were mainly attributed to the metabolism of arginine ，energy and nucleic acid. CONCLUSIONS：The anti tumor effect of C Ⅱ-3 may be related to the regulation of arginine metabolism ，energy metabolism and nucleic acid metabolism.

Effect of Different Processed Products of Crataegi Fructus on Dyspepsia by 1H-NMR Metabonomics / 中国实验方剂学杂志

Objective:The effect of processing on promoting digestion of Crataegi Fructus was investigated by the method of serum metabonomics based on 1H-NMR and the analysis of traditional biochemical indexes. Method:The dyspepsia model was induced by high protein and calorie diet. The small intestine propulsive rate in mice and the hormone level in rat gastrointestinal tract were used as pharmacodynamic indexes. After administration of Crataegi Fructus, fried and charred Crataegi Fructus, the changes of serum metabolites were analyzed by 1H-NMR technique combined with multivariate statistical analysis, so as to compare the therapeutic effects of each processed products on rats with dyspepsia model. Result:Each processed products group of Crataegi Fructus could improve the ability of intestinal propulsion and gastric emptying, especially in charred Crataegi Fructus group, caused by dyspepsia. The serum metabonomics proved that there were significant metabolic differences between the model group and the blank group. A total of 13 biomarkers related to dyspepsia, covering 3-hydroxybutyric acid, glycerophosphoryl choline (GPC), N-acetylglycoprotein, O-acetylglycoprotein, trimethylamine oxide (TMAO), alanine, acetic acid, glutamic acid, glutamine, creatine, leucine, lactic acid and glucose, were screened on the grounds of VIP value of S-plot, single factor ANOVA and area under receiver operating characteristic curve (ROC curve) (AUC). The metabolite composition of each administration group was the same except that the callback tendency of O-acetylglycoprotein, glutamine and GPC in charred Crataegi Fructus group was closer to that in the blank group. Conclusion:There are differences in the effect of promoting digestion to eliminate stagnation among processed products of Crataegi Fructus, charred Crataegi Fructus is more effective in improving dyspepsia, which may play a role in regulating gastrointestinal motility and energy metabolism.

Effect of Dry-cold Environment on Metabolic Components in Urine and Plasma of Rats / 中国实验方剂学杂志

Objective:Nuclear magnetic resonance hydrogen spectrum(1H-NMR) was adopted to investigate the effect of dry-cold environment and diet on urine and plasma metabolites of rats,in order to find possible biomarkers and their related metabolic pathways. Method:The dry-cold environment and diet were used as a stressors to intervene rats for 3 weeks,urine and plasma samples were collected.1H-NMR combined with multivariate data analysis were used to identify relevant metabolic markers and analyze their metabolic pathways. Result:The dry-cold environment and diet resulted in significant changes of 23 metabolites in urine and 15 metabolites in plasma;these metabolites were closely related to changes of various metabolic pathways,namely the tricarboxylic acid(TCA) cycle,pyruvate metabolism,glycolysis or gluconeogenesis,valine,leucine and isoleucine biosynthesis,glycine,serine and threonine metabolism,histidine metabolism;glyoxylic acid and dicarboxylic acid metabolism,alanine,aspartic acid and glutamate metabolism. Conclusion:Dry-cold environment and diet conditions can lead to the changes of various metabolic pathways in the body.The metabolomics based on 1H-NMR has the advantage of explaining the pathophysiological dynamics and overall changes of the body,which is of great significance for exploring the pathogenesis of non-deterministic disease factors.

1H-NMR based metabolomic study of sedative and hypnotic effect of Ziziphi Spinosae Semen in rats / 中草药

Objective: 1H-NMR based metabolomic approach has been employed to study the intervention effects of Ziziphi Spinosae Semen (ZSS) on p-chlorophenylalanine (PCPA)-induced insomnia using a rat model. Methods 1H-NMR analysis was used to identify the various compounds of ZSS, and the quantitative data were further processed by principal component analysis (PCA), partial least squares discrimination analysis (PLS-DA), and orthogonality PLS-DA to find potential biomarkers. In addition, MetPA analysis was used for pathway analysis. Results:Multivariate analysis revealed that 11 perturbed endogenous metabolites in the serum could be reversed by ZSS at dosage of 15 g/kg and 30 g/kg, and MetPA analysis revealed that the anti-hypnotic effect of ZSS was probably related with regulation of glutamate and glutamite metabolism, taurine and taurine metabolism, alanine, aspartic acid and glutamate metabolism, glyoxylic acid, and dicarboxylic acid metabolism, glycine, serine and threonine metabolism, pyruvate metabolism, tryptophan metabolism, and tricarboxylic acid cycle. ZSS at dosage of 15 g/kg and 30 g/kg both can significant callback 11 potential biomarkers. Conclusion: These metabolic changes indicated that ZSS facilitated recovery from insomnia by regulating the amino acid metabolism and energy metabolism, which provides the basis for elucidating therapeutic mechanisms underlying the efficiency of ZSS for insomnia treatment.

Effects of different processed products of Crataegi Fructus on hyperlipidemia rat model by 1H-NMR metabolomics / 中草药

Objective: To investigate the difference of lipid-lowering effect of raw and processed Crataegi Fructus, and to select the optimal antihyperlipidemic product. Methods The hyperlipidemia model of SD rats was induced by high fat diet. After administration of Crataegi Fructus (1.5 g/kg), fried and coked Crataegi Fructus (1.5 g/kg), the changes of serum metabolites were analyzed by 1H-NMR technique combined with multivariate statistical analysis. Results :Different processing products of Crataegi Fructus can significantly reduce the levels of TC, TG, LDL-C in hyperlipidemia rats, among which Crataegi Fructus had best hypolipidemic effects. The serum contents of lactate, pyruvate, lysine, creatine, glycerol, ornithine, TMAO, α-ketoisovaleric acid, O-acetylglycoprotein, N-acetylglycoprotein, leucine and isoleucine were increased in model group, while choline, acetone, threonine, scyllitol, glutamine, glutamic acid, MUFA, PUFA, GPC, and glucose were decreased compared with control group. After administration of different processing products of Crataegi Fructus, endogenous metabolites were recovered significantly. The effect of Crataegi Fructus on hyperlipidemia was superior to those of fried and coked Crataegi Fructus according to the degree and number of the metabolites recovered. Conclusion: There are differences between the raw and processed Crataegi Fructus, and raw Crataegi Fructus showed the best effect on hyperlipidemia, the effect may be achieved through regulating oxidative stress, energy metabolism, amino acid metabolism, and gut microbiota.

Chemical Comparison of the Stems and Leaves of Tussilago farfara L. Using NMR-based Metabolomics / 中国药学杂志

OBJECTIVE: To compare the chemical compositions of the stems (STL) and leaves (LTL) of Tussilago farfara L. using NMR-based metabolomic approach. METHODS: The STL and LTL were analyzed by NMR, then the differential components were determined by multivariate statistical METHODS:, including PCA, PLS-DA, OPLS-DA, and univariate analysis. RESULTS: Fifty compounds were tentatively identified in the NMR spectra of STL and LTL. Multivariate coupled with univariate analysis revealed that some metabolites, such as valine,leucine,isoleucine,proline,chlorogenic acid,3,5-O-dicaffeoylquinic acid, 3,4-O-dicaffeoylquinic acid, and tussilagone,were present at higher levels in the LTF, while the STF contained higher levels of α-glucose and β-glucose.For some metabolites, such as malic acid, sucrose and choline, no significant differences were observed between STF and LTF. CONCLUSION: This study reveals the chemical differences between STF and LTF in a holistic way, and lays the scientific foundation for the resource utilization of the stems and leaves of T. farfara L.