Comparison of alkaloids in Aconiti Kusnezoffii Radix, Aconiti Radix, and Aconiti Lateralis Radix Praeparata based on UHPLC-Q-Exactive Orbitrap MS/MS / 中国中药杂志

UHPLC-Q-Exactive Orbitrap MS/MS was used to systematically analyze and compare the alkaloids in Aconiti Kusnezoffii Radix, Aconiti Radix, and Aconiti Lateralis Radix Praeparata. After the samples were pretreated in the solid-phase extraction cartridges, 0.1% ammonium hydroxide(A)-acetonitrile(B) was used for gradient elution. The LC-MS method for characterization of alkaloids in the three herbal medicines was established in ESI positive ion mode to collect high resolution MS data of reference substances and samples. On the basis of the information of reference substance cracking behavior, retention time, accurate molecular mass, and related literature, a total of 155 alkaloids were identified in Aconiti Kusnezoffii Radix, Aconiti Radix, and Aconiti Lateralis Radix Prae-parata. Specifically, 130, 127, and 92 alkaloids were identified in Aconiti Kusnezoffii Radix, Aconiti Radix, and Aconiti Lateralis Radix Praeparata, respectively. Monoester alkaloids and amino-alcohol alkaloids were dominant in the three herbal medicines, and the alkaloids in Aconiti Kusnezoffii Radix and Aconiti Radix were similar. This paper can provide a reference for elucidating the pharmacological effects and clinical application differences of the three herbal medicines produced from plants of Aconitum.

Rapid identification of components in Wuzhuyu Decoction using UHPLC Q-Exactive Orbitrap MS~n and molecular network technology / 中国中药杂志

Wuzhuyu Decoction, the classical formula recorded in the Treatise on Febrile Diseases(Shang Han Lun), has been included in the Catalogue of Ancient Classic Prescriptions(the First Batch). Consisting of Euodiae Fructus, Ginseng Radix et Rhizoma, Zingiberis Rhizoma Recens, and Jujubae Fructus, it is effective in warming the middle, tonifying deficiency, dispelling cold, and descending adverse Qi, and is widely applied clinically with remarkable efficacies. For a classical formula, the chemical composition is the material basis and an important premise for quantity value transfer. This study aimed to establish a rapid identification method of chemical components in Wuzhuyu Decoction by high-resolution mass spectrometry(HR-MS) and molecular network. AQUITY UPLC BEH C_(18) column(2.1 mm×100 mm, 1.7 μm) was used for sample separation, and acetonitrile-0.1% formic acid in water was used as mobile phases for gradient elution. Q-Exactive Orbitrap MS data were collected in positive and negative ion modes, and GNPS molecular network was plotted according to the similarity of MS/MS fragmentation modes. Cytoscape 3.6.1 was used to screen molecular clusters with similar structures. Finally, the chemical components of Wuzhuyu Decoction were rapidly identified according to the controls, as well as the information of retention time, accurate relative molecular weight of HR-MS, and MS/MS multistage fragments. A total of 105 chemical components were identified in Wuzhuyu Decoction. This study can provide data for the follow-up quality control, standard substance research, and pharmacodynamic material research on Wuzhuyu Decoction, as well as references for the rapid qualitative analysis of the chemical components of Chinese medicine.

Qualitative and quantitative study of constituents in Lysionoti Herba based on UHPLC-Q-Exactive Orbitrap HRMS and HPLC-UV / 中国中药杂志

Ultra-high performance liquid chromatography-quadrupole-Exactive Orbitrap high resolution mass spectrometry(UHPLC-Q-Exactive Orbitrap HRMS) was employed to systematically analyze the chemical constituents in Lysionoti Herba, and high perfor-mance liquid chromatography-ultraviolet(HPLC-UV) to determine the content of main compounds. A Synergi~(TM) Hydro-RP 100 Å colu-mn(2 mm×100 mm, 2.5 μm) was used for gradient elution with acetonitrile-0.1% aqueous formic acid as the mobile phase at a flow rate of 0.2 mL·min~(-1) and a column temperature of 40 ℃. MS and MS/MS were conducted with electrospray ionization(ESI) in both positive and negative modes. The chemical components in Lysionoti Herba were identified by comparison with the retention time and mass spectra of reference compounds and the relevant mass spectral data reported in MS databases and relevant literature. Furthermore, the content of five constituents(neochlorogenic acid, chlorogenic acid, forsythoside B, acteoside, and nevadensin) in different Lysiono-ti Herba samples was simultaneously determined by HPLC-UV at the wavelength of 330 nm. A total of 84 compounds were identified in Lysionoti Herba, including 27 flavonoids, 20 phenylethanoid glycosides, 5 amino acids, 18 organic acids, 1 alkaloid, 6 nucleosides, and 7 others. The content of neochlorogenic acid, chlorogenic acid, forsythoside B, acteoside, and nevadensin showed good linear relationship(r>0.999) with the peak area within certain concentration ranges, which were 3.22-102.90, 12.84-410.82, 31.63-1 012.01, 25.00-800.11, and 4.08-130.51 μg·mL~(-1), respectively. The instrument precision, method repeatability, and solution stability all met requirement, and the average recovery rate was 97.31%-100.2%, with RSD ranging from 0.95% to 2.4%. The content of the five components varied among different Lysionoti Herba samples collected from different regions of Guizhou, and the average content of forsythoside B was the highest. The established qualitative method can rapidly and efficiently identify the chemical components of Lysionoti Herba, and the developed HPLC-UV method can simultaneously determine the content of five components in a simple, ra-pid, and accurate manner, providing a scientific basis for the quality evaluation of Lysionoti Herba.

Rapid identification of the chemical compounds in the leaves of Solanum nigrum L. based on UHPLC-Q Exactive MS and molecular network technology / 药学学报

The whole herb of Solanum nigrum L. can be used as the herbal drug. In this study, UHPLC-Q Exactive high resolution mass combined with GNPS molecular network was used for the rapid characterization of the components in the leaves of S. nigrum L. A total of 157 compounds were identified, including 30 steroid alkaloids, 61 steroid saponins, 35 flavonoids, and 31 other compounds (amino acids and organic acids), by comparison with the data reported in the literature, and mass fragmentation characteristics analysis, as well as the correlation of known and unknown nodes in the GNPS molecular network. Compared with the fruits and stems, the leaves of S. nigrum L was rich in a variety of steroidal saponins, steroidal alkaloids, and flavonoids, and the results lay the foundation for the precise resources utilization of S. nigrum L.

Neuroprotective effects of Longxue Tongluo Capsule on ischemic stroke rats revealed by LC-MS/MS-based metabolomics approach / 中草药·英文版

OBJECTIVE@#The present study aimed to evaluate the therapeutic effect and explore the underlying mechanisms of Longxue Tongluo Capsule (LTC) on ischemic stroke rats.@*METHODS@#Twenty-six rats were randomly divided into four groups, including sham group, sham + LTC group, MCAO group, and MCAO + LTC group. Ischemic stroke rats were simulated by middle cerebral artery occlusion (MCAO), and LTC treatment group were orally administrated with 300 mg/kg of LTC once daily for seven consecutive days. LTC therapy was validated in terms of neurobehavioral abnormality evaluation, cerebral infarct area, and histological assessments. The plasma metabolome comparisons amongst different groups were conducted by UHPLC-Q Exactive MS in combination with subsequent multivariate statistical analysis, aiming to finding the molecules in respond to the surgery or LTC treatment.@*RESULTS@#Intragastric administration of LTC significantly decreased not only the neurobehavioral abnormality scores but also the cerebral infarct area of MCAO rats. The interstitial edema, atrophy, and pyknosis of glial and neuronal cells occurred in the infarcted area, core area, and marginal area of cerebral cortex were improved after LTC treatment. A total of 13 potential biomarkers were observed, and Youden index of 11 biomarkers such as LysoPC, SM, and PE were more than 0.7, which were involved in neuroprotective process. The correlation and pathway analysis showed that LTC was beneficial to ischemic stroke rats via regulating glycerophospholipid and sphingolipid metabolism, together with nicotinate and nicotinamide metabolism. Heatmap and ternary analysis indicated the synergistic effect of carbohydrates and lipids may be induced by flavonoid intake from LTC.@*CONCLUSION@#The present study could provide evidence that metabolomics, as systematic approach, revealed its capacity to evaluate the holistic efficacy of TCM, and investigate the molecular mechanism underlying the clinical treatment of LTC on ischemic stroke.

Comparative chemical characters of Pseudostellaria heterophylla from geographical origins of China / 中草药·英文版

OBJECTIVE@#Pseudostellaria heterophylla has been paid more attention in recent years, mainly as a medicine food homology plant. The content determination of P. heterophylla is not specified in the Chinese Pharmacopoeia (version 2020). The environmental conditions in different production areas could exert an influence on the quality of P. heterophylla. The purpose of this study is to discriminate P. heterophylla collected from different geographical origins of China.@*METHODS@#In this study, the content of polysaccharide in 28 batches of P. heterophylla was determined using phenol-sulfuric acid. HPLC fingerprints were established under optimised HPLC-PDA methods. Subsequently, the similarity analysis (SA) and the quantification of heterophyllin B were analyzed. The metabolites of P. heterophylla were identified and evaluated using UHPLC-Q Exactive HF orbitrap MS system. Principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), hierarchical cluster analysis (HCA) and orthogonal PLS-DA (OPLS-DA) were performed based on all peak areas.@*RESULTS@#The polysaccharide content in Guizhou and Jiangsu was higher than that of other production areas, which varied significant from different origins. While the content of heterophyllin B in Anhui and Jiangsu was high. The correlation coefficients of HPLC fingerprints for 28 batches samples ranged from 0.877 to 0.990, and the characteristic map can be used to identify and evaluate the quality of P. heterophylla. The samples from Fujian, Guizhou, Jiangsu provinces can be relatively separated using multivariate statistical analysis including PCA, PLS-DA, HCA, OPLS-DA, indicating that their metabolic compositions were significantly different. Ultimately, a total of 15 metabolites which were filtrated by a VIP-value > 1 and a P-value < 0.05 associated with the separation of different origins were identified.@*CONCLUSION@#HPLC fingerprint was established to evaluate the quality and authenticity of P. heterophylla. The present work showed that the difference of geographic distributions had an influence on the internal chemical compositions. A sensitive and rapid untargeted metabolomics approach by UHPLC-Q Exactive HF orbitrap MS was utilized to evaluate P. heterophylla from different origins in China for the first time. Overall, this study provides insights to metabolomics of P. heterophylla and supplies important reference values for the development of functional foods.

Study on effective components of Ganoderma applanation and its mechanism of action in preventing COVID-19 based on UHPLC-Q- Exactive-Orbitrap-MS integrated network pharmacology / 中国药理学通报

Aim To elucidate the effective components of Ganoderma applanatum and its mechanism of preventing the coronavirus disease 2019(COVID-19).Methods To begin with, UHPLC-Q-Exactive-Orbitrap-MS was established to identify the main chemical constituents of G.applanatum.Then, the predicted targets of G.applanatum were selected by Swiss Target Prediction.GO analysis and KEGG analysis of core target genes were performed using the DAVID database.Finally, to explore the potential mechanism of G.applanatum against COVID-19, core functional components-core target-metabolism path network diagram was constructed using Cytoscape 3.8.0, and molecular docking was used to analyze the binding force of the core effective compounds with angiotensin-converting enzyme II(ACE2)and three SARS CoV-2 proteins, nonstructural protein-15 Endoribonuclease(NSP15), the receptor-binding domain of spike protein(RBD of S protein), and main protease(Mpro/3CLpro).Results Sixty-two components were identified from G.applanatum by UHPLC-Q-Exactive-Orbitrap-MS study; 30 active components were closely associated with 32 core targets including IL6, PTGS2, and MAPK1; KEGG analysis showed that it might treat COVID-19 through signaling pathways, such as PI3K-Akt signaling pathway, TNF signaling pathway, tuberculosis, and so on; molecular docking analysis showed that 1,4-Dihydroxy-2-naphthoic acid, parthenolide, 7,8-Dihydroxycoumarin, and other vital compounds had a certain degree of affinity with ACE2 and three SARS CoV-2 proteins.Conclusion This study clarifies the chemical composition and the potential mechanism of G.applanatum, providing a scientific basis for screening the effective ingredients of G.applanatum.

UHPLC-Q-Exactive Orbitrap MS/MS-based rapid identification of chemical components in substance benchmark of Kaixin San / 中国中药杂志

Ultra-performance liquid chromatography-quadrupole-electrostatic field Orbitrap mass spectrometry(UHPLC-Q-Exactive Orbitrap MS/MS) was used for rapid identification of the chemical components in Kaixin San substance benchmark. The gradient elution was performed through a Waters ACQUITY~(TM) BEH C_(18) column(2.1 mm×150 mm, 1.7 μm) with water-acetonitrile as mobile phase, a column temperature of 30 ℃, a flow rate of 0.3 mL·min~(-1), and a sample size of 1 μL. The scanning was performed in the negative ion mode. The complex component groups in Kaixin San substance benchmark were quickly and accurately identified and clearly assigned based on the comparison of the retention time and MS data with those of the reference substance as well as the relative molecular weight of the same or similar components in the mass spectrum database and literature. A total of 77 compounds were identified, including 26 saponins, 13 triterpenoid acids, 20 oligosaccharide esters, 5 xanthones, and 13 other compounds. The qualitative method established in this study can systematically, accurately, and quickly identify the chemical components in Kaixin San substance benchmark, which can provide a basis for the further analysis of its active components in vivo and the establishment of its quality control system.

Chemical analysis of classical prescription Qianghuo Shengshi Standard Decoction by UHPLC-Q Exactive Orbitrap MS / 中国中药杂志

A UHPLC-Q Exactive Orbitrap MS method was used to analyze the chemical constituents of the classical prescription Qianghuo Shengshi Standard Decoction(QHSS). UHPL conditions were as follows: Waters~(TM) UPLC~(TM) HSS T3 C_(18) column(2.1 mm×100 mm, 1.7 μm) and mobile phase of acetonitrile-0.1% formic acid aqueous solution. Mass spectrometry data of QHSS, each herb extract, and negative sample were collected in both positive and negative ion modes. The chemical constituents of QHSS were identified or tentatively identified based on the accurate molecular weight, retention time, MS fragmentation, comparison with reference substances, and literature reports. A total of 141 compounds were identified, including 18 amino acids, oligosaccharides, oligopeptides, and their derivatives, 19 phenolic acids, 44 coumarins, 18 flavonoids and chromones, 13 saponins, 17 phthalides, and 12 other components. This study comprehensively characterized the chemical constituents of QHSS, laying an experimental basis for the in-depth research on the material basis and quality control of QHSS.

Analysis on Chemical Constituents in Rhizomes of Bergenia scopulosa by UHPLC-Q Exactive Focus MS/MS / 中国实验方剂学杂志

Objective:Ultra-high performance liquid chromatography coupled with quadrupole/electrostatic field orbital trap high resolution mass spectrometry （UHPLC-Q Exactive Focus MS/MS） was developed to rapidly analyze and identify the chemical components in the rhizomes of <italic>Bergenia scopulosa</italic>. Method:The 75% methanol extract of <italic>B</italic>.<italic> scopulosa</italic> rhizomes was analyzed on a Thermo Accucore aQ RP18 column （2.1 mm×150 mm， 2.6 μm） with the mobile phase of methanol （A） and 0.1% formic acid aqueous solution （B） for gradient elution （0-40 min 5%-95%A， 40-45 min 95%A）， the flow rate was 0.3 mL·min^-1 and the column temperature was at 30 ℃. The information of the chemical constituents was acquired in positive and negative ion modes by heated electrospray ion source （HESI）， and the scanning range was <italic>m</italic>/<italic>z</italic> 80-1 200. Result:A total of 66 chemical constituents were identified， including 2 free amino acids， 7 bergenin derivatives， 15 flavonoids， 15 organic acids， 25 glycosides， and 2 others. Conclusion:The chemical constituents in the rhizomes of <italic>B</italic>.<italic> scopulosa</italic> can be identified systematically， accurately and rapidly by this method. Among them， 8 compounds were unambiguously identified by comparing with reference substances （succinic acid， arbutin， gallic acid， protocatechuic acid， bengenin， catechin， chlorogenic acid and caffeic acid）， 51 compounds were found from <italic>B</italic>.<italic> scopulosa</italic> for the first time and 28 compounds were found from the genus <italic>Bergenia</italic> for the first time. This paper can provide an important basis for the further material basis clarification and quality assessment of <italic>B</italic>.<italic> scopulosa</italic>.

Analysis of Metabolites and Metabolism Pathway of Tetrahydroxystilbene Glucoside in Rats with UHPLC-MS and Mass Defect Filter Method / 中国药房

OBJECTIVE：To analyze the metabolites of tetrahydroxystilbene glucoside （THSG）and speculate its metabolism pathway in rats. METHODS ：Male SD rats were randomly divided into plasma group （n＝3），urine group （n＝3），bile group （n＝3），and tissue group （n＝9）. Each group was given single dose of THSG 200 mg/kg intragastrically. Plasma samples 10，30 min and 1，1.5，2，4 h after medication ，the unrine 0-6 h after medication ，the bile 0-4 h after medication ，the tissue of heart ， liver，spleen，lung，kidney and stomach 30 min and 1，2 h after medication （3 at each time point ）were collected respectively.After precipitated with methanol ，the metabolites of samples were analyzed and identified by UHPLC-Q-Exactive Orbitrap MS and mass loss filtration （MDF）. Its metabolism pathway was speculated. RESULTS：In the blood ，urine，bile，heart，liver，spleen， lung，kidney，stomach samples ，6，7，11，1，5，1，3，4，4 metabolites were detected ，including two phase Ⅰ（hydrolysis， hydrogenation and hydroxylation ）metabolites，18 phase Ⅱ（glucuronic acid binding and sulfation ）metabolites. There were 12 glucuronic acid binding products. CONCLUSIONS：Most of the metabolites of THSG are found in bile ，mainly glucuronic acid binding products of phase Ⅱ metabolite THSG ； main metabolic pathways involve glucose hydrolysis ， hydrogenation， hydroxylation，glucuronic acid binding and sulfation.

UHPLC-Q-Exactive-Obitriap MS for quantification of ten components in Fuzi Lizhong Decoction / 中草药

Objective: To establish a method for quantifying ten components in Fuzi Lizhong Decoction (FLD). Methods: Ultra high performance liquid chromatography-Q-Exactive Obitriap mass spectrometry was used to detect components under optimized conditions as follows: Hypersil Gold C18 column (50 mm × 2.1 mm, 1.9 μm), column temperature: 30 ℃, mobile phase: methanol (A)-water (B), gradient elution conditions: 0-6 min, 25%-80% A, 6-7 min, 80%-95% A, 7-8 min, 95%-100% A, 8-9 min, 100% A, flow rate: 0.3 mL/min; Ion source: electrospray, scan mode: full scan (positive, negative), scan range: m/z 200-700 (positive), 200-500 (negative), resolution: 17 500, capillary temperature: 300 ℃, spray voltage: +4.0 kV, -3.5 kV, sheath gas flow rate: 35 L/h, S-lens voltage: 50 V. Results: Satisfactory linearity with correlation coefficient (r2) higher than 0.995 was achieved for each compound. The content of benzoyl aconitine, benzoylmesaconine, atractylenolide I, atractylenolide II and lobetyolin scanned under positive mode in FLD was 2.52, 0.11, 0.46, 1.75 and 5.8 μg/mL respectively, and that of emodin, glycyrrhizin, liguiritigenin, 8-gingerol, 10-gingerol scanned under negative mode in FLD was 0.35, 2.52, 0.98, 6.65 and 2.71 μg/mL, respectively. Conclusion: The developed liquid chromatography-mass spectrometry method was fastwith low limit of detection, which could provide methodological basis for quality control of FLD as well as quantification of these compounds.

Analysis of carnosic acid metabolites in rats by UHPLC-Q-Exactive MS / 中国中药杂志

A method of ultra-high performance liquid chromatography coupled with quadrupole/electrostatic field Obitrap high-resolution mass spectrometry(UHPLC-Q-Exactive MS) was established to comprehensively identify the metabolites of carnosic acid in rats. After oral gavage of carnosic acid CMC-Na suspension in rats, urine, plasma and feces samples were collected and pretreated by solid phase extraction(SPE). Acquity UPLC BEH C_(18 )column(2.1 mm×100 mm, 1.7 μm) was used with 0.1% formic acid solution(A)-acetonitrile(B) as the mobile phase for the gradient elution. Biological samples were analyzed by quadrupole/electrostatic field Obitrap high-resolution mass spectrometry in positive and negative ion mode. Based on the accurate molecular mass, fragment ion information, and related literature reports, a total of 28 compounds(including carnosic acid) were finally identified in rat samples. As a result, the main metabolic pathways of carnosic acid in rats are oxidation, hydroxylation, methylation, glucuronide conjugation, sulfate conjugation, S-cysteine conjugation, glutathione conjugation, demethylation, decarbonylation and their composite reactions. The study showed that the metabolism of carnosic acid in rats could be efficiently and comprehensively clarified by using UHPLC-Q-Exactive MS, providing a reference for clarifying the material basis and metabolic mechanism of carnosic acid.

Chemical profiling and tissue distribution study of Jingyin Granules in rats using UHPLC-Q-Exactive Orbitrap HR-MS / 中国中药杂志

In this study, the chemical profiling of Jingyin Granules and the tissue distribution of nine major constituents in this Chinese medicine were performed after oral administration of Jingyin Granules to rats, by using UHPLC-Q-Exactive Orbitrap HR-MS. An Acquity UPLC BEH C_(18) chromatographic column(2.1 mm×100 mm, 1.7 μm) was used as solid phase, while the mobile phase was methanol and 0.1% formic acid water for gradient elution. The major constituents in this Chinese medicine were quickly and accurately identified, via comparison with the retention times and MS/MS spectra of the standards. A total of 106 chemicals were identified from Jingyin Granules, including 24 kinds of organic acids, 47 kinds of flavonoids, 10 kinds of iridoids, and 21 kinds of saponins and 4 kinds of other compounds. After oral administered Jingyin Granules to rats, 48, 30, 25, 23, 45, 34, 39, 26, 19 prototype compounds were identified in serum, heart, liver, spleen, lung, kidney, brain, fat, and testicles, respectively. Meanwhile, an LC-MS based analytical method was established for simultaneous determination of chlorogenic acid, swertiamarin, caffeic acid, sweroside, liquiritin, prim-O-glucosylcimifugin, arctiin, 5-O-methylvisammioside and arctigenin in biological samples. The tissue distribution(serum, liver and lung) of these nine aim constituents in rats after oral administration of Jingyin Granules were investigated. It was found that these nine constituents could be quickly absorbed into circulation system and then distributed to liver and lung tissues. Except arctigenin, the exposure of other eight aim constituents to serum and lung was peaked at 1 h. At 1 h, the exposure of these components to lung tissue were ranked as follows: swertiamarin [(75 191.0±3 483.21) ng·g~(-1)]>arctiin [(2 716.5±36.06) ng·g~(-1)]>5-O-methylvisammioside [(585.1±0.71) ng·g~(-1)]>arctigenin [(437.45±3.18) ng·g~(-1)]>chlorogenic acid [(308.1±5.66) ng·g~(-1)]>prim-O-glucosylcimifugin [(211.35±2.19) ng·g~(-1)]>sweroside [(184.3±9.05) ng·g~(-1)]>caffeic acid [(175.95±2.05) ng·g~(-1)]>liquiritin [(174.78±153.34) ng·g~(-1)]. In summary, an UHPLC-Q-Exactive Orbitrap HR-MS method has been established for rapid and accurate identification of the constituents in Jingyin Granules, while the tissue distribution of nine major absorpted constituents were investigated in rats following oral administration of Jingyin Granules. These findings provided key information and guidance for further studies on pharmacodynamic substances and clinical applications of Jingyin Granules.

UHPLC-HRMS metabonomics to study effect of sulfur-fumigated Ophiopogonis Radix on endogenous metabolites in rats / 中国中药杂志

The project was launched to analyze the effects of sulfur-fumigated Ophiopogonis Radix on endogenous metabolites in rats by metabonomics. The preparation method of sulfur-fumigated Ophiopogonis Radix in laboratory was established. Then the blood samples of SD rats in blank group,Ophiopogonis Radix extract group and sulfur-fumigated Ophiopogonis Radix extract group were investigated by UHPLC-Q-Exactive. The differential metabolites were screened and identified by PCA(principal component analysis),OPLSDA(orthogonal partial least squares discriminant analysis) and variable importance projection(VIP),and the metabolic pathways were analyzed. Finally,a total of 15 potential biomarkers were identified. Compared with the samples of Ophiopogonis Radix extract group,sulfur-fumigated Ophiopogonis Radix mainly affected the biosynthesis and metabolism of amino acids in normal rats. Its mechanism may be related to the biosynthesis of phenylalanine,tyrosine,tryptophan and aminoacyl-tRNA as well as the metabolism of phenylalanine and tryptophan. Based on UHPLC-HRMS metabonomics,this paper discussed the effects of sulfur-fumigated Ophiopogonis Radix on endogenous metabolites in rats,which provided an idea for the metabolic study of other sulfur-fumigated traditional Chinese medicines.

Phospholipidomics study of Jiegeng Decotion for LPS-induced acute lung injury in mice based on UHPLC-Q-Exactive Orbitrap/MS / 药学学报

Using the lipidomics method based on UHPLC-Q-Exactive Orbitrap/MS, the change of phospholipid metabolism in lung tissue of mice induced by lipopolysaccharide (LPS)-induced acute lung injury was analyzed to observe the regulation of abnormal lipids by Jiegeng Decoction and to explore the regulation effect of Jiegeng Decoction on LPS-induced acute lung injury. The lung tissue samples from control group, model group, dexamethasone (positive drug) group, and Jiegeng Decoction group were collected and the lipid components of the sample were extracted. All procedures over mice were performed in accordance with the Guidelines for Care and Use of Laboratory Animals of Nanjing University of Chinese Medicine, and the experiments were approved by the Animal Ethics Committee of our university. The lipidomics technique of UHPLC-Q-Exactive Orbitrap/MS was used to study change of phospholipids in lung tissue of each group. LPS induced acute lung injury in mice with metabolic abnormalities of phospholipids, the specific performance of the PC was significantly upregulated, phosphatidyl ethanolamine (PE), phosphatidyl glycerol (PG), phosphatidyl serine (PS),phosphatidylinositol (PI) and other metabolic disorders, Jiegeng Decoction have a certain role in these phospholipids. LPS-induced acute lung injury caused disturbances of phospholipid in vivo, and Jiegeng Decoction regulates metabolic phospholipids.

Identification of major chemical constituents in Ixeris chinensis and Sonchus brachyotus by UHPLC-Q Exactive Orbitrap-HRMS / 中草药

Objective: To identify the similarities and differences of the main chemical constituents between Ixeris chinensis and Sonchus brachyotus by UHPLC-Q exactive orbitrap-HRMS. Methods: The analysis was performed on an Agilent Poroshell 120 reverse phase column (100 mm × 3 mm, 2.7 μm). The mobile phase consisted of methanol and 0. 5% acetic acid, which was used for gradient elution, and the flow rate was 0.3 mL/min. Q exactive orbitrap-HRMS spectrometry was applied for the qualitative analysis under positive and negative ion modes, and ESI ion source was used for mass spectra. Results: The results indicated that nine compounds from the ethanol extract of I. chinensis and ten compounds from the ethanol extract of S. brachyotus had been identified by direct comparison in both positive and negative ion mass data, the element compositions analysis, and the data of the literature. Among them, there were nine compounds are the same, which were seven organic acids and two flavonoids compounds. Conclusion: The efficient separation ability of UHPLC and high sensitive detection of MS were used in this study, which will provide the evidences for evaluating the quality of I. chinensis and S. brachyotus and stabilizing the curative effect in clinic.

Identification of chemical constituents in Naozhenning granule by UHPLC coupled with hybrid quadrupole-orbitrap MS / 药学学报

Naozhenning granule, which is composed of 11 herbal drugs, is mainly used in the treatment of concussion, cerebral post-traumatic syndrome. As the chemical composition of Naozhenning was complex, the ultrahigh performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry was applied to identify the chemical constituents rapidly in the Naozhenning granule. By analyzing the retention behaviors, accurate molecular weight, the fragmentation pathway, and comparison with the MS data from the standards and references, as well as the automatic identification by the Compound Discover 2.0, a total of 161 compounds were identified or tentatively deduced, and the sources of constituents were also confirmed. The compounds identified in Naozhenning granule included 9 iridoids, 8 butylphthalides, 26 flavonoids and flavonoid glycosides, 8 phenolic acids, 8 monoterpenoid glycosides and 9 alkaloids, as well as the common compounds in the herbal drugs, such as organic acids, amino acids, and sugars. The chemical composition of Naozhenning was studied for the first time, which provides a scientific basis for the quality control and document of effective materials of Naozhenning granule.

Rapid identification of chemical constituents in Huangqi Jianzhong Tang by UHPLC coupled with hybrid quadrupole-orbitrap MS / 药学学报

This study was designed to establish an ultra-liquid chromatography-mass spectrometry method for the reliable identification the multiple chemical components in Huangqi Jianzhong Tang (HQJZ). The ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry method was applied to identify the chemical constituents in the HQJZ rapidly. A total of 71 compounds including two major categories of saponins and flavonoids were identified or tentatively deduced on the basis of their retention behaviors, fragments of multistage mass spectrometry or by comparing with reference substances and literatures. Among them, 20 compounds were from Astragali Radix, 14 were from Paeoniae Radix Alba, 37 were from Glycyrrhizae Radix et Rhizoma Praeparata cum Melle, 3 were from Rhizoma Zingiberis Recens, 2 were from Cinnamomi Ramulus and Jujubae Fructus, respectively. The LC-MS method was used to qualitatively analyze the chemical constituents of HQJZ, which provides a scientific basis for the quality control of HQJZ.

Integrated strategy of UHPLC-Q Exactive Orbitrap-HRMS and HPLC-CL to study with constituents in Ziziphi Spinosae Folium / 中草药

Objective: To integrate UHPLC-Q Exactive Orbitrap-HRMS and HPLC-CL to study the major chemical constituents in Ziziphi Spinosae Folium (the leaves of Ziziphus jujuba var. spinosa) and to trace the active constituents on line. Methods: A UHPLC-Q Exactive Orbitrap-HRMS method was developed for the identification of multi-constituents in Ziziphi Spinosae Folium. The fingerprint for the multi-component assay in the leaves was established by HPLC-UV. The biological activity fingerprint of Ziziphi Spinosae Folium scavenging H2O2 and O2∙- was simultaneously identified for eight different areas in Shanxi. Results: Ten compounds in the leaves were identified or tentatively characterized. Among them, six flavones have scavenging activities for free radicals. Thirteen common peaks were calibrated and the correlation coefficients were greater than 0.90. Conclusion: A large number of flavones in Ziziphi Spinosae Folium have antioxidant activities. Ziziphi Spinosae Folium provides a novel, green, and sustainable source as antioxidant for the market.