Analytical Methods
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2024
·Vol. 16
·期 16
·2472-2477
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IF 2.7
Chenhao Zhao,Xingdan Wang,Jing Wu,Yeli Hu,Qiwei Zhang,Qi Zheng
Sialic acids are a family of monosaccharides that share a nine-carbon backbone and a carboxyl group. A recent derivatization method based on 3-nitrophenylhydrazine (3-NPH) provides a mild chemical labeling technique for biomolecules containing carbonyl or carboxyl groups. In this study, we utilized 3-NPH to label sialic acids via a two-step derivatization process. The derivatized species can produce a common reporter ion corresponding to C1–C3 with two labels, and a fragment differentiating between Neu5Ac, Neu5Gc, and KDN. This method is compatible with O-acetylated sialic acids and provides high sensitivity to Neu5Gc and KDN, and since the utilization of dual labeling significantly enhances the hydrophobicity of derivatives, it can effectively mitigate matrix effects when combined with parallel reaction monitoring technology. Negative-ion tandem mass spectrometry (MS/MS) analysis reveals a distinctive fragmentation profile for the 4-O-acetylated species, while the other sialic acids yield similar MS/MS spectra with a high abundance of reporter ions. Using the reporter ion as a transition, this analytical strategy is effective for analyzing complex biological samples. For example, it was successfully employed to quantify sialic acids in the intestinal tissues of several carp species, demonstrating its potential in sialylation research.
研究领域:
分析化学
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糖生物学
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PubMed
高影响因子
ACS Omega
·
2025
·
IF 4.3
Zhenghu Min,Xingdan Wang,Xiaoqiu Yang,Qiwei Zhang,Qi Zheng
Glycosylation profiling is an effective methodology for achieving a comprehensive understanding of glycoproteins and their alterations in a multitude of pathological conditions. However, in comparison to N-glycosylation, O-glycosylation presents significant challenges in terms of both qualitative and quantitative mass spectrometric analyses. A recently developed oxidative release protocol enables the selective formation of O-glycans containing a carboxyl group derived from the amino acid residue. In this study, 3-nitrophenylhydrazine was used to derivatize the common carboxyl group in a mild hydrophilic solution. Derivatization resulted in the generation of a series of report ions for serine, threonine, sialic acid, and O-acetylated sialic acid residues, thereby facilitating the identification of O-glycans and their attached amino acid residues, as well as the determination of the number of O-acetyl groups. A total of 65 O-glycans can be identified from bovine mucin. Furthermore, the analytical strategy revealed that O-acetylated N-acetylneuraminic acid (Neu5Ac)-containing O-glycans from horse serum exhibited distinctive fragmentation patterns in comparison to those from bovine mucin. Additionally, the presence of deaminoneuraminic acid (KDN)-containing O-glycans was successfully confirmed in fish intestinal tissue. These findings suggest that this method provides an economical and potentially valuable tool for large-scale O-glycosylation studies in complex biological samples.
研究领域:
生物化学
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蛋白质组学
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分析化学
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高影响因子
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
·
2024
·
IF 2.8
Qian Li,Yongfu Zeng,Lianfeng Ai,Maolin Wei,Xiaomeng Liu,Xiaochan Zhao,Haoran Zhang,Xiujuan Guo
A stable isotope dilution-liquid chromatography tandem mass spectrometry method based on a low-temperature derivatization strategy with 3-nitrophenylhydrazine (3-NPH) was developed for the determination of six volatile fatty acids (VFAs) in serum, urine, and feces. Ice acetonitrile was used to precipitate proteins and extract the target analytes. The extract was derivatized with 3-NPH methanol solution at 4 °C. BEH C8 (1.7 μm, 2.1 × 100 mm) column was used for chromatographic separation, and acetonitrile–water (both containing 0.01 % formic acid) were used as the mobile phase with a gradient elution of 10 min. Electrospray ionization source (ESI) in negative ion multiple reaction monitoring (MRM) mode were used for analyte detection. The regression coefficients R 2 of the calibration curves for the six VFAs were in the range of 0.9963–0.9994, and the LOQs were in the range of 0.02–0.5 μg mL −1 , with the recoveries in the range of 85.3–104.3 %, and the intra- and inter-day precision in the range of 1.8–9.1 %. The method is simple, accurate and reliable, and has been applied in the sensitive determination of VFAs in complex biological samples.
研究领域:
分析化学
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生物医学
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PubMed
高影响因子
Journal of Analysis and Testing
·
2020
·Vol. 4
·期 4
·249-256
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IF 0
Zuo Jiali,Cai Runxian,An Yanpeng,Tang Huiru
Abstract Stereoisomeric hexoses are present in almost all biological organisms in the forms of aldoses and ketoses, with diverse physiological and pathophysiological functions. Accurate and simultaneous quantification is vital for understanding their functions individually. However, such analysis remains challenging owing to their highly similar behavior in chromatography and mass spectrometry. By combining the pre-column 3-nitrophenylhydrazine derivatization and ultrahigh performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS), here, we developed a method for simultaneous quantification of five important stereoisomeric hexoses including D-glucose, D-galactose, D-mannose, D-fructose and L-sorbose representing both aldoses and ketoses. The method achieved baseline-separation for all these five derivatized hexoses chromatographically and had high sensitivity (LOD, femtomole on column), excellent linearity ( R 2 > 0.995) and efficiency with stable-isotope dilution. With this method, we further quantified these hexoses in nine biological matrices including human biofluids (serum, urine and saliva), human cells, human and mouse feces, rat liver tissue, mung-bean seeds and peach pulp. The results provided quantitative data for these hexoses in multiple biological samples and showed significant concentration diversity for these hexoses in different biological samples, which demonstrated the applicability of the method for simultaneous quantification of these hexose phenotypes of biological systems. Graphic Abstract
研究领域:
分析化学
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生物化学
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医学
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JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
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2018
·
IF 2.831
Lan Luo,Congcong Gu,Mingxian Li,Xiangyuan Zheng,Feng Zheng
4-Nitrobenzaldehyde is the synthetic raw material and an important photodegradation product of chloramphenicol . With a structural “alert” of human genotoxic potential and reported mutagenicity, this compound should be controlled in drug substances as a potential genotoxic impurity. However, current analysis methods require complex pre-treatment processes and/or lack sufficient specificity and sensitivity. Nitrophenylhydrazine is a common carbonyl derivatization reagent used to determine the residual aromatic aldehydes in drug samples. In the present study, we report an unexpected advantage of 3-nitrophenylhydrazine hydrochloride as a derivatization reagent in the derivatization high-performance liquid chromatography-ultraviolet detection method to determine 4-nitrobenzaldehyde in chloramphenicol samples. Compared with other nitro-substituted phenylhydrazines , 3-nitrophenylhydrazine hydrochloride can minimize drug matrix and derivatization reagent interferences, since the maximum absorption wavelength of its derivative is significantly red-shifted to 397 nm. The derivatization conditions have been optimized in terms of reaction efficiency, including reaction temperature, time, and diluting solvent, through a design of experiments. As a result, after reaction with 500 μg mL −1 of 3-nitrophenylhydrazine hydrochloride in acetonitrile-water (70:30, v/v) at 60 °C for 30 min, the developed HPLC method could be used to determine 4-nitrobenzaldehyde with a limit of detection of 0.009 μg mL −1 . The method was then validated and applied for the determination of residual 4-nitrobenzaldehyde in chloramphenicol and its eye-drop samples.
研究领域:
药物分析
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毒理学
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制药工程
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高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
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2018
·
IF 2.831
Ying-Ying Jin,Zi-Qi Shi,Wen-Qi Chang,Lin-Xiu Guo,Jian-Liang Zhou,Jian-Qun Liu,Li-Fang Liu,Gui-Zhong Xin
Recent studies have demonstrated the important role of short-chain fatty acids (SCFAs) in the maintenance of homeostasis of respiratory immunity. However, there is still no report focus on the determination of SCFAs level in bronchoalveolar lavage fluid (BALF), the most common sample used for screening biomarkers of the pulmonary diseases. Herein, an ultra-high-performance liquid chromatography with LTQ-Orbitrap mass spectrometer (UHPLC-LTQ-Orbitrap) oriented 3-nitrophenylhydrazine (3-NPH)-based derivatization method was developed for the quantification of SCFAs in BALF. To achieve accurate quantitation, d4-acetate was used as internal standard to compensate for the matrix effects. Method validation showed a good linearity ( R 2 > 0.9992) with wide concentration range, and the intra-day and inter-day precision for determination of eight SCFAs in BALF samples was ≤ 14.79%. The quantitation accuracy, assessed by relative recoveries, ranged from 90% to 110% for target SCFAs at three concentration levels. Matrix effects ranged from 85% to 115%, and the lower limits of quantification of these targeted SCFAs were varied from 3 to 24 nmol/L. The SCFAs-targeted method was then applied to determine the changed levels in BALF samples from OVA-induced asthma mice and normal mice. In addition, the universality of our developed method was also demonstrated by determining the SCFAs concentrations in feces, serum and lung tissue samples from asthma and normal mice. These results indicate that 3-NPH derivatization based UHPLC-LTQ-Orbitrap provides accurate view of global SCFAs alternation in different samples, giving a support to deduce the origin of SCFAs in lung. The present study is of great importance for understanding the role of SCFAs in modulation of host metabolism and immunity.
研究领域:
分析化学
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呼吸医学
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免疫学
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质谱技术
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高影响因子
Foods
·
2025
·Vol. 14
·期 21
·
IF 5.1
Junxia Ma,Yingshan Jiang,Yue Teng,Ting Ren,Yanchao Xing,Aoyun Li,Zhongmei He,Weijia Chen,Ying Zong,Rui Du
Rheumatoid arthritis (RA) is a chronic autoimmune inflammatory disease, and current treatments are limited by significant side effects. Deer bone, which is rich in proteins and various active compounds, possesses anti-inflammatory and bone-health-promoting properties. However, its fermented product’s effects on RA treatment remain unexplored. In this study, we evaluated the therapeutic effects of probiotic-fermented deer bone aqueous extract (BbF) in an adjuvant arthritis (AA) rat model, combined with LPS-stimulated RAW264.7 macrophage models. In vivo experiments showed that BbF significantly reduced paw swelling, arthritis index, and improved bone mineral density. BbF also alleviated synovial hyperplasia and inflammatory cell infiltration. It suppressed pro-inflammatory cytokines (TNF-α, IL-1β, etc.) and inhibited macrophage migration and invasion. Transcriptomic analysis revealed significant enrichment of the Notch signaling pathway, and Western blot confirmed the downregulation of Notch3, Notch4, DLL4, and Jagged1 proteins. BbF also restored gut microbiota homeostasis, increasing beneficial bacteria such asFirmicutesandClostridia, while decreasing potential pathogens likeProteobacteria,Gammaproteobacteria, andEscherichia-Shigella. Furthermore, BbF enhanced short-chain fatty acids (SCFCs) production, including butyrate and caproic acid. These results indicate that BbF alleviates RA by inhibiting the Notch signaling pathway and regulating gut microbiota, providing new insights for the development of functional foods with immune-modulatory properties.
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Journal of Inflammation Research
·
2025
·
IF 4.1
Yuan Zhang,Longbin Huang,Ningning Yue,Zhiliang Mai,Chen Kong,Chengmei Tian,Dao-ru Wei,Jun Yao,Lisheng Wang,Defeng Li
Background Although cytokines have been implicated in the development of ulcerative colitis (UC), the potential mediating role of metabolite levels in this association remains unclear.Methods Utilizing data from genome-wide association studies (GWAS) encompassing 91 circulating cytokines, 1400 blood metabolites, and 178,689 UC cases, we performed a two-sample Mendelian randomization (MR) analysis to investigate the effect of metabolites mediated cytokines on the development of UC. A two-step MR analysis was conducted to quantitatively evaluate the mediation effect. Additionally, dextran sodium sulfate (DSS)-induced colitis mice were used to further confirm our results.Results Mendelian randomization (MR) analysis indicated that macrophage colony-stimulating factor (M-CSF) had a causal and positive relationship with aconitate (OR: 1.10, 95% CI: 1.00–1.20, p = 0.043, IVW beta 1 = 0.095). Moreover, MR analysis revealed that high level of aconitate were associated with reduced risk of UC (OR: 0.44, 95% CI: 0.24–0.80, p = 0.008, IVW beta 2 = −0.818). In addition, MR analysis showed M-CSF had an inverse correlation with the disease onset of UC (OR: 0.31, 95% CI: 0.15–0.80, p = 0.002, IVW beta all = −1.16). Furthermore, the mediation effect of aconitate mediated M-CSF on the risk of UC was −0.0777 (95% CI: −0.154 to −0.0018, p = 0.045), accounting for 6.69% of the total effect, and indicating a modest contribution to the protective effect of M-CSF against UC. Subsequently, as an in vivo validation model, DSS-induced colitis was employed to demonstrate that M-CSF treatment significantly ameliorated weight loss, disease activity index (DAI) scores, colon shortening, and histological damage. Additionally, M-CSF treatment also significantly reduced M1 macrophage infiltration, elevated levels of aconitate as well as itaconate, and decreased the levels of pro-inflammatory cytokines in colitis. These results demonstrated that aconitate inhibited the expression of pro-inflammatory cytokines through its enzymatic conversion into the immunometabolite itaconate by aconitate decarboxylase 1 (Acod1), and downregulated the levels of M1 macrophages, thereby ameliorating colitis.Conclusion These findings suggest that M-CSF is an important anti-inflammatory cytokine in UC, which may be a promising therapeutic target in the treatment of UC.
研究领域:
免疫学
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胃肠病学
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遗传学
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代谢组学
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炎症研究
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生物医学研究
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PubMed
高影响因子
ACS Nano
·
2024
·
IF 15.8
Yanling Wu,Zhenping Bao,Songhao Zhang,Rui Liu,Yuan Ping,Min Ma,Yuxia Gao,Chengyi He,Tianyue Wu,Yongqiang Ma,Chenhui Zhang,Fengpei Du
Agrichemical losses are a severe threat to the ecological environment. Additionally, some agrichemical compounds contain abundant salt, which increases the instability of formulations, leading to a lower agrichemical utilization and soil hardening. Fortunately, the biological amphiphilic emulsifier sodium deoxycholate alleviates these problems by forming stable Janus core–shell emulsions through salinity-driven interfacial self-assembly. According to the interfacial behavior, dilational rheology, and molecular dynamics simulations, Janus-emulsion molecules are more closely arranged than traditional-emulsion molecules and generate an oil–water interfacial film that transforms into a gel film. In addition, at the same spray volume, the deposition area of the Janus emulsion increased by 37.70% compared with that of the traditional emulsion. Owing to the topology effect and deformation, the Janus emulsion adheres to rice micropapillae, achieving better flush resistance. Meanwhile, based on response of the Janus emulsion to stimulation by carbon dioxide (CO2), the emulsion lost to the soil can form a rigid shell for inhibiting the release of pesticides and metal ions from harming the soil. The pyraclostrobin release rate decreased by 50.89% at 4 h after the Janus emulsion was exposed to CO2. The Chao1 index of the Janus emulsion was increased by 12.49% as compared to coconut oil delivery in soil microbial community. The Janus emulsion ingested by harmful organisms can be effectively absorbed in the intestine to achieve better control effects. This study provides a simple and effective strategy, which turns waste into treasure, by combining metal ions in agrichemicals with natural amphiphilic molecules to prepare stable emulsions for enhancing agrichemical rainfastness and weakening environmental risk.
研究领域:
农业化学
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环境科学
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材料科学
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生物技术
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PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2025
·
IF 8.5
Nan Ma,Rong Li,Gui-Fang Zhang,Ruo-Han Gao,Dong-Jie Zhang
Quinoa β-glucan (QBG) has shown potential benefits in treating type 2 diabetes mellitus (T2DM); however, comprehensive evaluations of its effects remain limited. This study investigates the impact of QBG—derived from hot water extraction (Q-) and microbial fermentation enrichment (Q+)—on serum glucose levels, lipid profiles, appetite-regulating hormones, fecal short-chain fatty acids (SCFAs), and gut microbiota composition and function in streptozotocin/high-fat diet (STZ/HFD)-induced T2DM mice. The results indicate that QBG treatment significantly reduced fasting blood glucose, insulin levels, triglycerides (TG) and total cholesterol (TC), while concurrently increasing high-density lipoprotein cholesterol (HDL C) levels. Additionally, liver and pancreatic function improved, as evidenced by decreased levels of malondialdehyde (MDA), aspartate transaminase (AST), and alanine transaminase (ALT). SCFA levels were significantly higher in QBG-treated groups compared to MC group. QBG treatment also reduced the abundance of Firmicutes and Patescibacteria, along with the Firmicutes/Bacteroidota ratio, while increasing levels of Bacteroidota and Actinobacteria. These findings suggest that QBG can regulate the dysbiosis of SCFAs production in T2DM mice and may indirectly modulate the secretion of appetite-regulating hormones by influencing gut microbiota composition. Furthermore, PICRUSt analysis revealed that QBG treatment, particularly Q + _H, could enhance disrupted metabolism and improve gut microbiota functions, helping restore normal physiological function.
研究领域:
营养学
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微生物学
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代谢生物学
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医学
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PubMed
高影响因子
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