scholarly journals Isolation of chloroplasts from marine microalgae Isochrysis galbana Parke suitable for organelle lipid composition analysis

2020 ◽  
Author(s):  
Ye Zhong ◽  
Yanrong Li ◽  
Jilin Xu ◽  
Jiayi Cao ◽  
Chengxu Zhou ◽  
...  
Keyword(s):  
Lipid A ◽  
Lipid Synthesis ◽  
Isochrysis Galbana ◽  
Fatty Acyl ◽  
Marine Microalgae ◽  
Composition Analysis ◽  
Biochemical Assays ◽  
Liquid Chromatography Tandem Mass ◽  
Fatty Acid Proportion ◽  
Ultrahigh Performance Liquid Chromatography

Abstract Background: Marine microalgae, Isochrysis galbana Parke, is an important diet microalgal species with high nutritional value. Different from other unicellular microalgae, its cell contains two chloroplasts which are the major sites for lipid synthesis. Results: Here, we optimized a chloroplast isolation approach suitable for I. galbana Parke, and evaluated the purity and integrity of the isolated chloroplasts by microscopic observations and biochemical assays. The chloroplast lipids were sequenced by ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Glycolipids were the main chloroplast lipids, and DGDG, MGDG, and SQDG were the most abundant glyceroglycolipids. DGMG and SQMG were not observed among the chloroplast lipids. In addition, DG was the most abundant neutral lipid. A part of fatty acyl R1/R2 with MGMGs, DGDGs, MGDGs, SQDGs, PEs, and PCs were not found in chloroplasts. The fatty acid proportion of chloroplast lipids were increased, decreased, or remained unchanged compared with the whole-cell. Conclusions: This newly developed isolation approach was a simple and reliable method to isolate chloroplasts with high integrity and purity. Collectively, our findings show that such an isolation approach may be used in studies on many different aspects of chloroplast biology, and offers a useful reference for isolation of chloroplast from other marine microalgae.

scholarly journals Dilution of QuEChERS Extracts Without Cleanup Improves Results in the UHPLC-MS/MS Multiresidue Analysis of Pesticides in Tomato

2021 ◽  
Author(s):  
Fabiane M. Stringhini ◽  
Lucila C. Ribeiro ◽  
Graziela I. Rocha ◽  
Juliana D. de B. Kuntz ◽  
Renato Zanella ◽  
...  
Keyword(s):  
Matrix Effect ◽  
Practical Method ◽  
Analysis Method ◽  
Multiresidue Analysis ◽  
Limit Of Quantification ◽  
Chromatography Tandem Mass Spectrometry ◽  
The Matrix ◽  
Liquid Chromatography Tandem Mass ◽  
Ultrahigh Performance Liquid Chromatography ◽  
Positive Results

AbstractTomato is well-known to be one of the most cultivated and consumed vegetables worldwide and frequently contain pesticide residues. Therefore, a simple multiresidue method was established and validated to determine 129 pesticides and metabolites in tomato samples using a modified acetate QuEChERS without cleanup for sample preparation and determination by ultrahigh-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). Dilution of the raw extract in different proportions of mobile phase was evaluated and a dilution of 10 times presented adequate results improving analysis performance while minimizing the matrix effect. Validation performed according to SANTE guideline presented satisfactory results. Practical method limit of quantification was 0.01 mg kg−1 for most compounds. Recoveries between 70 and 120% with precision ≤ 20% were found for most compounds and spike levels evaluated. Matrix effect results were not significant for most part of compounds. Method proved to be simple, robust, and effective to be applied in routine analysis. Method applicability was performed by analysis of samples commercialized in Brazil and positive results were found demonstrating the importance of the proposed method.

scholarly journals Development and Application of an UHPLC-MS/MS Method for Comparative Pharmacokinetic Study of Eight Major Bioactive Components from Yin Chen Hao Tang in Normal and Acute Liver Injured Rats

2018 ◽  
Vol 2018 ◽  
pp. 1-12 ◽  
Author(s):  
Yun Wang ◽  
Xinrui Xing ◽  
Yan Cao ◽  
Liang Zhao ◽  
Sen Sun ◽  
...  
Keyword(s):  
Lower Limit ◽  
Limit Of Detection ◽  
Injury Model ◽  
Relative Standard ◽  
Limit Of Quantitation ◽  
Liquid Chromatography Tandem Mass ◽  
Bioactive Ingredients ◽  
Aloe Emodin ◽  
Plasma Pharmacokinetics ◽  
Ultrahigh Performance Liquid Chromatography

Yin Chen Hao Tang (YCHT) is one of the most famous hepatoprotective herbal formulas in China, but its pharmacokinetic investigation in model rats has been rarely conducted. In this study, the hepatic injury model was caused by intraperitoneal injections of carbon tetrachloride (CCl4), and YCHT was orally administered to the model and normal rats. An ultrahigh performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) method was established to analyze the plasma pharmacokinetics of eight major bioactive ingredients from YCHT in both the normal and liver injured rats. The calibration curves presented good linearity (r > 0.9981) in the concentration range. The relative standard deviation (RSD%) of inter- and intraday precision was within 9.55%, and the accuracy (RE%) ranged from -10.72% to 2.46%. The extraction recovery, matrix effect, and stability were demonstrated to be within acceptable ranges. The lower limit of detection (LLOD) and lower limit of quantitation (LLOQ) were around 0.1 ng/mL and 0.5 ng/mL, respectively, which were much lower than those in other related researches. Results reveal that there are significant differences in the pharmacokinetics of scoparone, geniposide, rhein, aloe-emodin, physcion, and chrysophanol in hepatic injured rats as compared to those in control except for scopoletin and emodin. Our experimental results provide a meaningful reference for the clinical dosage of YCHT in treating liver disorders, and the improvement of LLOD and LLOQ can also broaden the range of our method’s application, which is very suitable for quantitating these eight compounds with low levels.

scholarly journals Interactions between gastric microbiota and metabolites in gastric cancer

2021 ◽  
Vol 12 (12) ◽  
Author(s):  
Daofeng Dai ◽  
Yan Yang ◽  
Jieqing Yu ◽  
Tianfeng Dang ◽  
Wenjing Qin ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Amino Acids ◽  
16S Rrna Gene Sequencing ◽  
Rrna Gene ◽  
Metabolome Analysis ◽  
Tumor Tissues ◽  
Liquid Chromatography Tandem Mass ◽  
Ultrahigh Performance Liquid Chromatography ◽  
Rrna Gene Sequencing ◽  
Gastric Microbiota

AbstractThe development and progression of gastric cancer (GC) is greatly influenced by gastric microbiota and their metabolites. Here, we characterized the gastric microbiome and metabolome profiles of 37 GC tumor tissues and matched non-tumor tissues using 16s rRNA gene sequencing and ultrahigh performance liquid chromatography tandem mass spectrometry, respectively. Microbial diversity and richness were higher in GC tumor tissues than in non-tumor tissues. The abundance of Helicobacter was increased in non-tumor tissues, while the abundance of Lactobacillus, Streptococcus, Bacteroides, Prevotella, and 6 additional genera was increased in the tumor tissues. The untargeted metabolome analysis revealed 150 discriminative metabolites, among which the relative abundance of the amino acids, carbohydrates and carbohydrate conjugates, glycerophospholipids, and nucleosides was higher in tumor tissues compared to non-tumor tissues. The targeted metabolome analysis further demonstrated that the combination of 1-methylnicotinamide and N-acetyl-D-glucosamine-6-phosphate could serve as a robust biomarker for distinction between GC tumors and non-tumor tissues. Correlation analysis revealed that Helicobacter and Lactobacillus were negatively and positively correlated with the majority of differential metabolites in the classes of amino acids, carbohydrates, nucleosides, nucleotides, and glycerophospholipids, respectively, suggesting that Helicobacter and Lactobacillus might play a role in degradation and synthesis of the majority of differential metabolites in these classes, respectively. Acinetobacter, Comamonas, Faecalibacterium, Sphingomonas, and Streptococcus were also significantly correlated with many differential amino acids, carbohydrates, nucleosides, nucleotides, and glycerophospholipids. In conclusion, the differences in metabolome profiles between GC tumor and matched non-tumor tissues may be partly due to the collective activities of Helicobacter, Lactobacillus, and other bacteria, which eventually affects GC carcinogenesis and progression.

scholarly journals Phosphoglycolipid Profiling of Bacterial Endotoxins

2018 ◽  
Vol 46 (1) ◽  
pp. 7-11 ◽  
Author(s):  
Anikό Kilár ◽  
Ágnes Dornyel ◽  
Viktor Sándor ◽  
Ferenc Kilár ◽  
Béla Kocsis
Keyword(s):  
Lipid A ◽  
De Novo ◽  
Clinical Symptoms ◽  
Phosphorylation Site ◽  
Reversed Phase ◽  
Fatty Acyl ◽  
Structural Elucidation ◽  
Bacterial Endotoxins ◽  
Product Ions ◽  
Tandem Mass Spectrometric

Abstract Much interest is at present focused on bacterial endotoxins, also known as lipopolysaccharides (LPS), as they are responsible for the development of clinical symptoms of Gram-negative sepsis which is the leading cause of death in intensive care units. Endotoxicity is associated with the special phosphoglycolipid part of LPS, termed lipid A. Main challenges in the structural elucidation of lipid A arise from its amphiphilic character and inherent heterogeneity. A mass spectrometrybased de novo method combined with reversed-phase liquid chromatography for the detailed structural characterization of complex lipid A mixtures (obtained by mild acid hydrolysis of LPS) from different bacterial sources has been developed. Tandem mass spectrometric measurements were performed with an electrospray-ionisation quadrupole time-of-flight (ESI-Q-TOF) mass spectrometer in both negative- and positive-ionization modes in order to explore fragmentation pathways. It was found that characteristic product ions in the positive-ion mode could be used for the unambiguous assignment of the phosphorylation site, whereas the use of both ionization modes provided consistent and/or complementary information about the fatty acyl distribution between the two glucosamine moieties of lipid A. Since the immunostimulatory (advantageous) vs. proinflammatory (endotoxic) effect of the lipid A is closely related to the fine chemical structure, our relatively simple structural elucidation strategy could offer great potential in the bioanalysis of native lipid A samples and lipid A-based vaccines

Effect of TiO 2 nanoparticle aggregation on marine microalgae Isochrysis galbana

2018 ◽  
Vol 66 ◽  
pp. 208-215 ◽  
Author(s):  
Ji Hu ◽  
Jianmin Wang ◽  
Shuxia Liu ◽  
Zhechao Zhang ◽  
Haifeng Zhang ◽  
...  
Keyword(s):  
Isochrysis Galbana ◽  
Marine Microalgae ◽  
Nanoparticle Aggregation

scholarly journals Comparative Study on Pharmacokinetics of Four Active Compounds in Rat Plasma after Oral Administration of Raw and Wine Processed Chuanxiong Rhizoma

Molecules ◽  
2019 ◽  
Vol 25 (1) ◽  
pp. 93 ◽  
Author(s):  
Yan Ning ◽  
Ke Pei ◽  
Gang Cao ◽  
Hao Cai ◽  
Xiao Liu ◽  
...  
Keyword(s):  
Oral Administration ◽  
Negative Ions ◽  
Rat Plasma ◽  
Phase System ◽  
Exact Mass ◽  
Active Compounds ◽  
Liquid Chromatography Tandem Mass ◽  
Ultrahigh Performance Liquid Chromatography ◽  
Switching Mode ◽  
T Values

In Chinese medicine, the effect of promoting blood circulation and removing stasis could be enhanced after Chuanxiong Rhizoma is processed by wine. However, the relevant mechanism remains unclear. In this manuscript, a rapid and sensitive quantification method employing ultrahigh performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was established and validated to simultaneously determine butylidenephthalide, ligustilide, senkyunolide A and ferulic acid in rat plasma after oral administration of raw Chuanxiong Rhizoma (RCR) and wine-processed Chuanxiong Rhizoma (WCR) respectively. All analytes were extracted from plasma by proteins precipitation with methanol. Chromatographic separation was carried out on a Hypersil GOLD C18 column by using a gradient mobile phase system of acetonitrile and water with 0.01% formic acid, the flow rate was 0.3 mL/min. For exact mass detecting, quick switching mode was used, positive and negative ions could be detected in one injection. The pharmacokinetic profiles of four components in the two groups were evaluated and compared. The results showed that, compared to the RCR group, the Vd and AUC0→t values of four active compounds were increased and decreased respectively in WCR group, which revealed the effect of wine processing to Chuanxiong Rhizoma: the stronger the effect, the wider the distribution.

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