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19 pages, 2159 KiB

Open AccessFeature PaperArticle

Protein Extraction, Enrichment and MALDI MS and MS/MS Analysis from Bitter Orange Leaves (Citrus aurantium)

by Donatella Aiello, Carlo Siciliano, Fabio Mazzotti, Leonardo Di Donna, Roberta Risoluti and Anna Napoli

Molecules 2020, 25(7), 1485; https://doi.org/10.3390/molecules25071485 - 25 Mar 2020

Cited by 21 | Viewed by 3733

Abstract

Citrus aurantium is a widespread tree in the Mediterranean area, and it is mainly used as rootstock for other citrus. In the present study, a vacuum infiltration centrifugation procedure, followed by solid phase extraction matrix-assisted laser desorption ionization tandem mass spectrometry (SPE MALDI [...] Read more.

Citrus aurantium is a widespread tree in the Mediterranean area, and it is mainly used as rootstock for other citrus. In the present study, a vacuum infiltration centrifugation procedure, followed by solid phase extraction matrix-assisted laser desorption ionization tandem mass spectrometry (SPE MALDI MS/MS) analysis, was adopted to isolate proteins from leaves. The results of mass spectrometry (MS) profiling, combined with the top-down proteomics approach, allowed the identification of 78 proteins. The bioinformatic databases TargetP, SignalP, ChloroP, WallProtDB, and mGOASVM-Loc were used to predict the subcellular localization of the identified proteins. Among 78 identified proteins, 20 were targeted as secretory pathway proteins and 36 were predicted to be in cellular compartments including cytoplasm, nucleus, and cell membrane. The largest subcellular fraction was the secretory pathway, accounting for 25% of total proteins. Gene Ontology (GO) of Citrus sinensis was used to simplify the functional annotation of the proteins that were identified in the leaves. The Kyoto Encyclopedia of Genes and Genomes (KEGG) showed the enrichment of metabolic pathways including glutathione metabolism and biosynthesis of secondary metabolites, suggesting that the response to a range of environmental factors is the key processes in citrus leaves. Finally, the Lipase GDSL domain-containing protein GDSL esterase/lipase, which is involved in plant development and defense response, was for the first time identified and characterized in Citrus aurantium. Full article

(This article belongs to the Special Issue Advanced Mass Spectrometry in Vegetable, Animal, and Human Qualitative and Quantitative Metabolomics)

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11 pages, 2785 KiB

Open AccessArticle

Chemical Discrimination of Astragalus mongholicus and Astragalus membranaceus Based on Metabolomics Using UHPLC-ESI-Q-TOF-MS/MS Approach

by Yumei Wang, Lei Liu, Yukun Ma, Lina Guo, Yu Sun, Qi Liu and Jicheng Liu

Molecules 2019, 24(22), 4064; https://doi.org/10.3390/molecules24224064 - 09 Nov 2019

Cited by 32 | Viewed by 3603

Abstract

Astragalus mongholicus (MG) and Astragalus membranaceus (MJ), both generally known as Huangqi in China, are two perennial herbals widely used in variety diseases. However, there were still some differences in the chemical ingredients between MG and MJ. In this paper, metabolomics combined with [...] Read more.

Astragalus mongholicus (MG) and Astragalus membranaceus (MJ), both generally known as Huangqi in China, are two perennial herbals widely used in variety diseases. However, there were still some differences in the chemical ingredients between MG and MJ. In this paper, metabolomics combined with the ultra-high performance liquid chromatography coupled with electrospray ionization/quadrupole time-of-flight mass spectrometry (UHPLC-ESI-Q-TOF-MS/MS) was employed to contrastively analyze the chemical constituents between MG and MJ. As a result, principal component analysis showed that MG and MJ were separated clearly. A total of 53 chemical markers were successfully identified for the discrimination of MG and MJ. Of them, the contents of 36 components including Astragaloside I~III, Astragaloside IV, Agroastragaloside I, etc. in MJ were significantly higher than those in MG. On the contrary, the contents of 17 other components including coumaric acid, formononetin, sophoricoside, etc. in MG were obviously higher than those in MJ. The results showed that the distinctive constituents in MG and MJ were remarkable, and MJ may own stronger pharmacological activities than MG. In a word, MG and MJ may be treated as two different herbs. This paper demonstrated that metabolomics was a vitally credible technology to rapidly screen the characteristic chemical composition of traditional Chinese medicine. Full article

(This article belongs to the Special Issue Advanced Mass Spectrometry in Vegetable, Animal, and Human Qualitative and Quantitative Metabolomics)

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21 pages, 5154 KiB

Open AccessArticle

Identification of Auxin Metabolites in Brassicaceae by Ultra-Performance Liquid Chromatography Coupled with High-Resolution Mass Spectrometry

by Panagiota-Kyriaki Revelou, Maroula G. Kokotou and Violetta Constantinou-Kokotou

Molecules 2019, 24(14), 2615; https://doi.org/10.3390/molecules24142615 - 18 Jul 2019

Cited by 10 | Viewed by 4626

Abstract

Auxins are signaling molecules involved in multiple stages of plant growth and development. The levels of the most important auxin, indole-3-acetic acid (IAA), are regulated by the formation of amide and ester conjugates with amino acids and sugars. In this work, IAA and [...] Read more.

Auxins are signaling molecules involved in multiple stages of plant growth and development. The levels of the most important auxin, indole-3-acetic acid (IAA), are regulated by the formation of amide and ester conjugates with amino acids and sugars. In this work, IAA and IAA amide conjugates with amino acids bearing a free carboxylic group or a methyl ester group, along with some selected IAA metabolites, were studied in positive and negative electrospray ionization (ESI) modes, utilizing high-resolution mass spectrometry (HRMS) as a tool for their structural analysis. HRMS/MS spectra revealed the fragmentation patterns that enable us to identify IAA metabolites in plant extracts from eight vegetables of the Brassicaceae family using a fast and reliable ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QToF-MS) method. The accurate m/z (mass to charge) ratio and abundance of the molecular and fragment ions of the studied compounds in plant extracts matched those obtained from commercially available or synthesized compounds and confirmed the presence of IAA metabolites. Full article

(This article belongs to the Special Issue Advanced Mass Spectrometry in Vegetable, Animal, and Human Qualitative and Quantitative Metabolomics)

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11 pages, 960 KiB

Open AccessArticle

Development and Validation of a Method for the Analysis of Bisoprolol and Atenolol in Human Bone

by Lucia Fernandez-Lopez, Manuela Pellegrini, Maria Concetta Rotolo, Aurelio Luna, Maria Falcon and Rosanna Mancini

Molecules 2019, 24(13), 2400; https://doi.org/10.3390/molecules24132400 - 29 Jun 2019

Cited by 10 | Viewed by 2793

Abstract

A method based on gas chromatography–mass spectrometry (GC–MS) is described for the determination of bisoprolol and atenolol in human bone. After the addition of lobivolol as internal standard, pulverized samples were incubated in acetonitrile for 1 h under ultrasounds. After adjusting the pH [...] Read more.

A method based on gas chromatography–mass spectrometry (GC–MS) is described for the determination of bisoprolol and atenolol in human bone. After the addition of lobivolol as internal standard, pulverized samples were incubated in acetonitrile for 1 h under ultrasounds. After adjusting the pH of the samples to 6, they were centrifuged, and the supernatants were subjected to solid phase extraction. Elution was achieved by using 3 mL of 2% ammonium hydroxide in 80:20 dichloromethane:isopropanol solution. Eluted samples were evaporated and derivatized. Chromatography was performed on a fused silica capillary column and analytes were determined in the selected-ion-monitoring (SIM) mode. The assay was validated in the range 0.1–0.3 ng/mg (depending on the drug) to 150 ng/mg, the mean absolute recoveries were 60% for bisoprolol and 106% for atenolol, the matrix effect was 69% for bisoprolol and 70% for atenolol and process efficiency was 41% for bisoprolol and 80% for atenolol. The intra- and inter-assay accuracy values were always better than 12%. The validated method was then applied to bone samples from two real forensic cases in which toxicological analysis in blood were positive for atenolol in the first case (0.65 µg/mL) and bisoprolol in the second case (0.06 µg/mL). Atenolol was found in bone samples from the corresponding case at the approximate concentration of 148 ng/mg and bisoprolol was found at 8 ng/mg. Full article

(This article belongs to the Special Issue Advanced Mass Spectrometry in Vegetable, Animal, and Human Qualitative and Quantitative Metabolomics)

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9 pages, 1497 KiB

Open AccessArticle

Dissipation Dynamics and Dietary Risk Assessment of Kresoxim-Methyl Residue in Rice

by MingNa Sun, Lu Yu, Zhou Tong, Xu Dong, Yue Chu, Mei Wang, TongChun Gao and JinSheng Duan

Molecules 2019, 24(4), 692; https://doi.org/10.3390/molecules24040692 - 15 Feb 2019

Cited by 5 | Viewed by 2477

Abstract

Kresoxim-methyl is a high-efficiency and broad-spectrum fungicide used for the control of rice fungal diseases; however, its residues after application potentially threaten human health. Investigations on the dissipation of kresoxim-methyl residue in rice field systems and dietary risk assessment of kresoxim-methyl in humans [...] Read more.

Kresoxim-methyl is a high-efficiency and broad-spectrum fungicide used for the control of rice fungal diseases; however, its residues after application potentially threaten human health. Investigations on the dissipation of kresoxim-methyl residue in rice field systems and dietary risk assessment of kresoxim-methyl in humans are limited. The present study employed the QuEChERS-GC-MS/MS method for residue analysis of kresoxim-methyl in rice plants, brown rice, and rice husks. The samples were extracted with acetonitrile and purified by PSA, C₁₈ column, and GCB. The average recovery of the spiked target compounds in the three matrices was between 80.5% and 99.3%, and the RSD was between 2.1% and 7.1%. The accuracy and precision of the method is in accordance with the requirements of residue analysis methods. Dissipation dynamic testing of kresoxim-methyl in rice plants indicated a half-life within the range of 1.8–6.0 days, and a rapid dissipation rate was detected. Dietary intake risk assessment showed that the national estimated daily intake (NEDI) of kresoxim-methyl in various Chinese subpopulations was 0.022–0.054 μg/(kg bw·days), and the risk quotient (RQ) was 0.0000055–0.00014%. These findings indicate that the risk for chronic dietary intake of kresoxim-methyl in brown rice is relatively low. The present study provides information and theoretical basis for guiding the scientific use of kresoxim-methyl in rice fields and evaluating its dietary risk in brown rice. Full article

(This article belongs to the Special Issue Advanced Mass Spectrometry in Vegetable, Animal, and Human Qualitative and Quantitative Metabolomics)

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13 pages, 2350 KiB

Open AccessArticle

A Microbial Transformation Model for Simulating Mammal Metabolism of Artemisinin

by Yue Ma, Peng Sun, Yifan Zhao, Kun Wang, Xiaoqiang Chang, Yue Bai, Dong Zhang and Lan Yang

Molecules 2019, 24(2), 315; https://doi.org/10.3390/molecules24020315 - 16 Jan 2019

Cited by 16 | Viewed by 3243

Abstract

Artemisinin (ART) is a highly effective antimalarial agent isolated from the traditional Chinese herb Qinghao. Metabolism of ART and its derivatives in the body is one of the most pressing issues for pharmaceutical scientists. Herein, an efficient in vitro microorganism model for simulation [...] Read more.

Artemisinin (ART) is a highly effective antimalarial agent isolated from the traditional Chinese herb Qinghao. Metabolism of ART and its derivatives in the body is one of the most pressing issues for pharmaceutical scientists. Herein, an efficient in vitro microorganism model for simulation of metabolism of ART in vivo was developed employing Cunninghamella elegans. Metabolites in the microbial transformation system and plasma of mice pre-administrated ART orally were analyzed by ultra-performance liquid chromatography (UPLC)-electrospray ionization (ESI)-quadrupole time-of-flight (Q-TOF)-mass spectrometry (MS^E) combined with UNIFI software. Thirty-two metabolites were identified in vitro and 23 were identified in vivo. After comparison, 16 products were found to be common to both models including monohydroxylated ART, dihydroxylated ART, deoxyartemisinin, hydroxylated deoxyartemisinin, hydroxylated dihydroartemisinin (DHA), and hydroxylated deoxy-DHA. These results revealed that C. elegans CICC 40250 functioned as an appropriate model to mimic ART metabolism in vivo. Moreover, an overall description of metabolites of ART from C. elegans CICC 40250 has been provided. Notably, DHA was detected and identified as a metabolite of ART in mouse plasma for the first time. Full article

(This article belongs to the Special Issue Advanced Mass Spectrometry in Vegetable, Animal, and Human Qualitative and Quantitative Metabolomics)

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21 pages, 3602 KiB

Open AccessArticle

Characterization and Quantification of Polyphenols and Triterpenoids in Thinned Young Fruits of Ten Pear Varieties by UPLC-Q TRAP-MS/MS

by Liqiong Sun, Shutian Tao and Shaoling Zhang

Molecules 2019, 24(1), 159; https://doi.org/10.3390/molecules24010159 - 03 Jan 2019

Cited by 64 | Viewed by 4741

Abstract

Large quantities of thinned young pears, a natural source of bioactive compounds, are abandoned as agricultural by-products in many orchards. Hence, ten thinned young pear varieties were systematically investigated in terms of their chemical composition and antioxidant potential. Through ultra-performance liquid chromatography coupled [...] Read more.

Large quantities of thinned young pears, a natural source of bioactive compounds, are abandoned as agricultural by-products in many orchards. Hence, ten thinned young pear varieties were systematically investigated in terms of their chemical composition and antioxidant potential. Through ultra-performance liquid chromatography coupled with electrospray ionization triple quadrupole mass spectrometry (UPLC-Q TRAP-MS/MS), 102 polyphenols and 16 triterpenoids were identified and individually quantified within a short time using multiple reaction monitoring (MRM). Subsequently, the antioxidant capacities of these pears were determined with DPPH assays, and the correlation between total antioxidant activity and each component was analyzed. The results indicated that the bioactive compound content and antioxidant capacity in thinned pears were considerably high. Regarding chemical composition, chlorogenic acid, quinic acid and arbutin were the primary polyphenols and ursolic acid was the predominant triterpenoid, whereas 27 polyphenolic compounds, especially chlorogenic acid and most of the flavan-3-ols, were the main antioxidants in young pears. These findings should provide a scientific basis for the further use of pear fruit by-products. Full article

(This article belongs to the Special Issue Advanced Mass Spectrometry in Vegetable, Animal, and Human Qualitative and Quantitative Metabolomics)

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16 pages, 4739 KiB

Open AccessArticle

Identification and Pharmacokinetic Studies on Complanatuside and Its Major Metabolites in Rats by UHPLC-Q-TOF-MS/MS and LC-MS/MS

by Yu-Feng Yao, Chao-Zhan Lin, Fang-Le Liu, Run-Jing Zhang, Qiu-Yu Zhang, Tao Huang, Yuan-Sheng Zou, Mei-Qi Wang and Chen-Chen Zhu

Molecules 2019, 24(1), 71; https://doi.org/10.3390/molecules24010071 - 25 Dec 2018

Cited by 7 | Viewed by 3306

Abstract

The metabolic and pharmacokinetic studies on complanatuside, a quality marker of a Chinese materia medicatonic, Semen Astragali Complanati, were carried out. The UHPLC-Q-TOF/MS (ultra-high performance liquid chromatography coupled with electrospray ionization tandem quadrupole-time-of-flight mass spectrometry) method was applied to identify the metabolites [...] Read more.

The metabolic and pharmacokinetic studies on complanatuside, a quality marker of a Chinese materia medicatonic, Semen Astragali Complanati, were carried out. The UHPLC-Q-TOF/MS (ultra-high performance liquid chromatography coupled with electrospray ionization tandem quadrupole-time-of-flight mass spectrometry) method was applied to identify the metabolites of complanatuside in rat plasma, bile, stool, and urine after oral administration at the dosage of 72 mg/kg. Up to 34 metabolites (parent, 2 metabolites of the parent drug, and 31 metabolites of the degradation products) were observed, including processes of demethylation, hydroxylation, glucuronidation, sulfonation, and dehydration. The results indicated glucuronidation and sulfonation as major metabolic pathways of complanatuside in vivo. Meanwhile, a HPLC-MS method to quantify complanatuside and its two major metabolites—rhamnocitrin 3-O-β-glc and rhamnocitrin—in rat plasma for the pharmacokinetic analysis was developed and validated. The T_max (time to reach the maximum drug concentration) of the above three compounds were 1 h, 3 h, and 5.3 h, respectively, while the C_max (maximum plasma concentrations)were 119.15 ng/mL, 111.64 ng/mL, and 1122.18 ng/mL, and AUC(0-t) (area under the plasma concentration-time curve) was 143.52 µg/L·h, 381.73 µg/L·h, and 6540.14 µg/L·h, accordingly. The pharmacokinetic characteristics of complanatuside and its two metabolites suggested that complanatuside rapidly metabolized in vivo, while its metabolites—rhamnocitrin—was the main existent form in rat plasma after oral administration. The results of intracorporal processes, existing forms, and pharmacokinetic characteristics of complanatuside in rats supported its low bioavailability. Full article

(This article belongs to the Special Issue Advanced Mass Spectrometry in Vegetable, Animal, and Human Qualitative and Quantitative Metabolomics)

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