TALANTA
·
2021
·
IF 6.057
Ning Sheng,Hongyi Zhao,Xiong Chen,Dongmei Wang,Menglin Li,Zhe Wang,Jinlan Zhang,Jiandong Jiang
Phosphate esters and anhydrides have great significance in the field of biochemical research and medical therapy. The genetic materials (DNA or RNA), most of the coenzymes , many intermediary metabolites, such as nucleotides and glycosyl phosphates in vivo are phosphodiesters, phosphoric acid or phosphates, respectively. It is important to monitor endogenous active phosphate metabolites for investigating many biological processes or drug mechanism. However, the detection and determination of those free active phosphate metabolites are challenged due to their unstable and easily hydrolyzed property and relatively low sensitivity, especially diphosphates and triphosphates. In the current study, we successfully developed a strategy by 3-aminomethyl pyridine (AMPy) derivatization coupled with hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) for simultaneous determination of multiple types of phosphate metabolites with good stability in 48 h and 29 to 126-fold improvement of the limit of detection (LOD). Based on the diagnostic fragment ions of different types of AMPy-derivatized phosphate metabolites, characteristic MRM ion pairs were successfully performed for global profiling of the phosphate metabolites in phosphate ester/anhydride metabolic network, including nucleotide/deoxynucleotide mono/di/triphosphates, glycosyl mono/diphosphates, and other key phosphates, such as 5-phosphoribosyl-1-pyrophosphate (PRPP), SAICARP and FAICARP in HPF, HUVEC and PBMCs cells without standards. The developed strategy greatly expanded the coverage of applying a single derivatization reaction to analyze active phosphate metabolites. Finally, the established method was performed to investigate the phosphate esters and anhydrides based on a glioma rat model. For the first time, phosphate metabolites were comprehensively characterized based on phosphate ester and anhydride metabolic network, covering nucleotide metabolism, glycolysis and pentose phosphate pathways, etc . The results demonstrated that the applicability of the method could be extended to a wider range of active phosphate compounds and could facilitate to related applications in the future studies.
研究领域:
生物化学
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医学治疗
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分析化学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF FOOD COMPOSITION AND ANALYSIS
·
2023
·
IF 4.3
Ran Xin,Meng Dong,Yu-Ying Zhang,Xu-Hui Huang,Xiu-Ping Dong,Lei Qin
Taste-active compounds are significant for the taste and nutrition of food, but their simultaneous quantification remains challenging due to the lack of efficient methods. The existing methods are difficult to simultaneously quantify many species and quantities of taste-active compounds by a single assay. In this study, a rapid and reliable method based on hydrophilic interaction chromatography coupled with triple quadrupole-tandem mass spectrometry (HILIC-MS/MS) was established for the simultaneous quantification and validation of 51 taste-active compounds in different food samples. The established method had a wide linear range (0.1–3000 ng/mL) and the limits of quantification were within the range from <0.1 to 100 ng/mL. This method demonstrated good quantification recovery in the range of 60–130 %. With the method proposed, the simultaneous determination of 51 taste-active compounds in 25 food samples was successfully applied. The quantitative and comparative results of taste-active compounds in different food products reveal the chemical characteristics and differences among different types of foods. This method showed good performance and a good application prospect for the simultaneous quantification of taste-active compounds in food samples. The analysis of key taste-active compounds in various food raw materials can enrich the diversification and individual flavor customization needs of food flavorings.
研究领域:
分析化学
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食品科学
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质谱分析
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DOI
高影响因子
Journal of Pharmaceutical Analysis
·
2026
·
IF 8.9
Yangrui Zhang,Yizhen Liu,Yangyang Chen,Chen Guo,Fengting Ou,Junhuan Lin,Hanmeng Guo,Tao Ke,Lushan Yu
The molecular complexity of active pharmaceutical ingredients (APIs) has increased over recent decades, with production largely relying on microbial processes such as fermentation. However, these processes introduce non-therapeutic protein impurities. Accurate quantification of protein impurities in APIs is critical, as underestimation risks safety and efficacy, while overestimation leads to costly over-purification. Here, we evaluated five conventional protein assays with diverse standards and found their accuracy compromised by protein structural diversity. To deconstruct the mechanism of this limitation, we examined the Bradford assay as a case study. Machine learning models trained on data from over 140 standards revealed that, beyond the known roles of basic and aromatic residues, other amino acids exert positive or negative effects on the Bradford assay response—as predicted by Lasso regression and validated with five designed peptides. Together with molecular docking and interaction analysis, which identified electrostatic interactions as the primary driving force, these findings collectively account for the inherent bias of such structure-dependent assays. The systematic evaluation and mechanistic insight prompted us to develop a hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry (HILIC-MS/MS) method for amino acid analysis, enabling accurate protein quantification at ppm levels in matrices. Applied to eight commercial APIs, the method revealed substantial variation in protein residue levels (9.53–5570 ppm). By providing precise measurement of protein impurities in small-molecule APIs, this study helps ensure drug safety and efficacy at no excessive purification costs, while also offering insights into the molecular mechanisms underlying the Bradford assay.
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DOI
高影响因子
ANALYTICAL SCIENCES
·
2023
·
IF 1.6
Lai Liang,Zhang Mengyun,Liu Chusheng,Qu Jiahuan,Xu Dongsheng,Jiang Zhengjin
Rapid and effective separation of nucleotides (NTs) and their derivatives is crucial for studying their physiological functions. In this work, we comprehensively evaluated the separation ability of a zwitterionic hydrophilic monolith, i.e., poly(N,N-dimethyl-N-(3-methacrylamidopropyl)-N-(3-sulfopropyl)ammonium betaine- co –N,N’-methylenebisacrylamide) (poly(SPP- co -MBA)) for NTs analysis, including its selectivity, chemical stability under extremely basic condition and compatibility with hydrophilic interaction liquid chromatography (HILIC) coupled with mass spectrometry (HILIC-MS). The poly(SPP- co -MBA) monolith exhibited excellent chemical stability, as evidenced by the low relative standard deviation of retention time (0.16–1.05%) after 4000 consecutive injections over one month under strong alkaline elution condition (pH 10). After optimizing the separation conditions, including buffer pH and concentration, organic solvent content and column temperature, four nucleoside triphosphates, five nucleoside diphosphates and five nucleoside monophosphates were baseline separated within 7 min. Additionally, the mixtures containing one nucleoside and its corresponding mono-, di-, and triphosphates were baseline separated within only 3 min, respectively. It is good HILIC-MS compatibility was also confirmed by the satisfactory peak shape and high response of nine NTs. Overall, the proposed poly(SPP- co -MBA) monolith exhibited good mechanical stability and compatibility of HILIC-MS, making it a promising technique for NTs analysis. Graphical Abstract
研究领域:
分析化学
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生物化学
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DOI
PubMed
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
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2021
·
IF 3.935
Ya Zhang,Qian Zhou,Xin Ding,Haibo Wang,Guangguo Tan
Doxorubicin (DOX) is widely used against cancer but carries the risk of a progressive cardiomyopathy. Astragalus polysaccharides (ASP) is the main active ingredient of Astragalus membranaceus Bunge. It has been proved to be effective against DOX-induced cardiomyopathy. However, its therapeutic mechanism is not yet well explored. In this study, a metabolomics approach based on hydrophilic interaction liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (HILIC-Q-TOFMS) was developed to characterize the metabolic fluctuations associated with DOX cardiomyopathy and elucidate the underlying mechanisms of the protective effects of ASP. By the combination of HILIC-Q-TOFMS and multivariate and univariate data analysis, we identified 22 polar serum metabolites associated with DOX cardiomyopathy, 12 of which were significantly reversed when the animals were co-treated with ASP through two main metabolic pathways, i.e., sphingolipid and glycerophospholipid metabolism. Furthermore, it was found that ASP could alleviate DOX-induced cardiomyopathy by decreasing the levels of acid sphingomyelinase, acid ceramidase and phospholipase A2 and increasing the levels of sphingomyelin synthase to regulate the sphingolipid and glycerophospholipid metabolic disorder. These results revealed that sphingolipid and glycerophospholipid metabolism may be significantly responsible for DOX cardiomyopathy, which is also a major mechanism for the action of ASP against DOX cardiomyopathy.
研究领域:
药理学
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心脏病学
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代谢组学
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DOI
PubMed
高影响因子
FITOTERAPIA
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2024
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IF 2.5
He Sun,Jia Lou,Bo-Xue Chen,Jia-Qi Huang,Qi-Long Wang,Shao-Fei Song,Zi-Yue Jia,Rong Miao,Shi-Yu Wang,Xue Li,Wen-Zhi Yang
Prunella vulgaris (PV) is a medicine and food homologous plant, but its quality evaluation seldom relies on the polysaccharides (PVPs). In this work, we established the multi-level fingerprinting and in vitro anti-inflammatory evaluation approaches to characterize and compare the polysaccharides of P. vulgaris collected from the major production regions in China. PVPs prepared from 22 batches of samples gave the content variation of 5.76–24.524 mg/g, but displayed high similarity in the molecular weight distribution. Hydrolyzed oligosaccharides with degrees of polymerization 2–14 were characterized with different numbers of pentose and hexose by HILIC-MS. The tested 22 batches of oligosaccharides exhibited visible differences in peak abundance, which failed to corelate to their production regions. All the PVPs contained Gal, Xyl, and Ara, as the main monosaccharides . Eleven batches among the tested PVPs showed the significant inhibitory effects on NO production on LPS-induced RAW264.7 cells at 10 μg/mL, but the exerted efficacy did not exhibit correlation with the production regions. Conclusively, we, for the first time, investigated the chemical features of PVPs at three levels, and assessed the chemical and anti-inflammatory variations among the different regions of P. vulgaris samples.
研究领域:
生药学
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分析化学
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药理学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
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2022
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IF 3.571
Zhenyao Lu,Sijia Li,Nan Aa,Yuanmao Zhang,Ran Zhang,Chen Xu,Shize Zhang,Xiangqing Kong,Guangji Wang,Jiye Aa,Yue Zhang
Purine and pyrimidine metabolism are vital metabolic pathways in the development, proliferation or repairment of cells or tissues associated with various diseases. Here, a simple, all-in-one injection hydrophilic interaction liquid chromatography-tandem mass spectrometry method was developed for simultaneous determination of 20 metabolites: adenine , adenosine, deoxyadenosine , adenosine 5′-monophosphate, cyclic adenosine monophosphate, hypoxanthine , xanthine , inosine , deoxyinosine , xanthosine , xanthosine 5′-monophosphate and uric acid , which are products of purine metabolism; uridine , deoxyuridine , uridine 5′-monophosphate and uracil , are products of pyrimidine metabolism; and corticosterone , methionine , acetylcholine and serotonin. To minimize interference of endogenous molecules in sample matrixes, a combination of activated carbon adsorption and a serum substitute matrix (5% bovine serum albumin in phosphate buffered saline) was utilized and jointly applied. The sensitivity, linearity, stability, precision, accuracy and extraction recovery were evaluated, and the method was demonstrated to be accurate, sensitive and reliable. An analytical strategy was successfully applied to quantitatively determine 20 metabolite levels in the serum and hippocampus of mice with chronic social defeat stress-induced depression. The results showed greatly perturbed purine metabolism in the depressed mice, which was primarily characterized by dramatic increases in hypoxanthine, xanthine and inosine in serum and reduced levels of adenine, adenosine and adenosine 5′-monophosphate in the hippocampus. These findings suggest that this novel strategy can facilitate the quantitative analysis of adenine and other purine and pyrimidine metabolites in tissue and serum and exhibits great potential in the exploration of metabolism-related mechanisms of relevant diseases.
研究领域:
生物化学
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医学
·
神经科学
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DOI
高影响因子
Foods
·
2024
·Vol. 13
·期 5
·635
·
IF 4.7
Aiping Li,Chen Wang,Zhenghao Wu,Yingying Liu,Zhenxia Hao,Chengyin Lu,Hongping Chen
Ningnanmycin is a widely used antibiotic in agricultural production that effectively controls fungal and viral diseases in tea trees and chrysanthemums. The polarity characteristic of ningnanmycin has posed limitations on the development of robust detection methods, thereby hindering effective monitoring and control measures. By combining cation exchange solid phase extraction (SPE) with hydrophilic interaction chromatography tandem mass spectrometry (HILIC-MS/MS), we have effectively tackled the issue pertaining to the separation and retention of ningnanmycin. The average recoveries of ningnanmycin in green tea, black tea, and chrysanthemum were 77.3–82.0%, 80.1–81.5%, and 74.0–80.0%, respectively. The intraday and interday relative standard deviations (RSDs) were below and equal to 7.7%. Good linearity was observed in the concentration range of 1–1000 μg/L (R2 > 0.998). The limits of detection (LODs) ranged from 1.1 μg/kg to 7.1 μg/kg, and the limits of quantification (LOQs) ranged from 3.6 μg/kg to 23.7 μg/kg for ningnanmycin. These results indicate the good accuracy, repeatability, reproducibility, and sensitivity of the method. It is suitable for detecting ningnanmycin in tea and chrysanthemum.
研究领域:
分析化学
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食品科学
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植物保护
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产品关联
DOI
PubMed
高影响因子
CARBOHYDRATE POLYMERS
·
2023
·
IF 11.2
Zhenhua Li,Xiaoqin Wang,Xiaoli Deng,Jiansen Song,Tong Yang,Yujie Liao,Guiping Gong,Linjuan Huang,Yu Lu,Zhongfu Wang
Glycosphingolipids (GSLs) in human milk regulate the immune system, support intestinal maturation, and prevent gut pathogens. The structural complexity and low abundance of GSLs limits their systematic analysis. Here, we coupled the use of monosialoganglioside 1–2-amino-N-(2-aminoethyl) benzamide (GM1-AEAB) derivatives as internal standards with HILIC-MS/MS to qualitatively and quantitatively compare GSLs in human, bovine, and goat milk. One neutral glycosphingolipid (GB) and 33 gangliosides were found in human milk, of which 22 were newly detected and three were fucosylated. Five GB and 26 gangliosides were identified in bovine milk , of which 21 were newly discovered. Four GB and 33 gangliosides were detected in goat milk, 23 of them newly reported. GM1 was the main GSL in human milk; whereas disialoganglioside 3 (GD3) and monosialogangloside 3 (GM3) were dominant in bovine and goat milk, respectively; N -acetylneuraminic acid (Neu5Ac) was detected in >88 % of GSLs in bovine and goat milk. N -hydroxyacetylneuraminic acid (Neu5Gc)-modified GSLs were 3.5 times more abundant in goat than in bovine milk; whereas GSLs modified with both Neu5Ac and Neu5Gc were 3 times more abundant in bovine than in goat milk. Given the health benefits of different GSLs, these results will facilitate the development of custom-designed human milk-based infant formula.
研究领域:
营养学
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生物化学
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代谢组学
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DOI
PubMed
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
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2020
·
IF 9.038
Fei Long,Mei Zhang,Jiahao Yuan,Juan Du,Ande Ma,Jialiang Pan
The measurement of trace hydrophilic toxins in complex aqueous-rich matrices is a daunting challenge. To address this analytical bottleneck, pulse diffusion focusing (PDF), a novel sample injection technique for hydrophilic interaction chromatography-tandem mass spectrometry (HILIC-MS/MS), was developed. Theoretical and experimental investigations of the mechanism and key parameters revealed that the pulse-injection approach, assisted by solvent diffusion, efficiently solved the volume overload problem. This milliliter-level-injection HILIC-MS/MS technique was reported for the first time herein, and provided a significant enhancement in sensitivity compared to the conventional injection method, in addition to being an efficient approach to address the solvent incompatibility of off-line sample preparation and HILIC . The automated PDF-HILIC-MS/MS system was obtained by slightly modifying a commercial LC–MS/MS instrument in an easy and economical manner. The efficiency of the system was demonstrated through the detection of trace tetrodotoxin contents in plasma and urine samples. Low limits of detection (i.e., 0.65 and 2.2 ng·mL −1 ) were obtained using the simplified sample preparation method. The recoveries were in the range 91–113.3 % with intra-day and inter-day precisions of ≤9.6 %. Further experimental results proved the method to be versatile for various hydrophilic toxins.
研究领域:
分析化学
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毒理学
·
医学检测
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DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2026
·
IF 6.7
Xiaoqin Wang,Yujie Liao,Xiaoxiao Guo,Daohang Xing,Xiang Chen,Qian Wang,Linjuan Huang,Zhongfu Wang
Bovine lactoferrin (bLf) is an essential additive in infant formula, with its bioactivity closely related to N-glycosylation. Currently, the production cost of lactoferrin remains high. This study compared eight ion-exchange resins and identified DoWex and 732 as optimal for bLf purification. Yields reached 4.46% for DoWex and 5.25% for 732, with purities exceeding 85% for both resins. Isomer-specific N-glycan analysis using HILIC-MS/MS revealed 58 distinct N-glycans in mature bLf, including 28 neutral and 30 acidic glycans, with the acidic N-glycans predominantly modified by α2,6-linked Neu5Ac. The bLf prepared by 732 resin shared >97% of the neutral and >93% of the acidic glycans with that of DoWex. Notably, the price of 732 resin is only one-18th that of DoWex, representing a significant reduction in production costs and making it promising in future large-scale manufacturing. This work facilitates efficient lactoferrin production and provides further studies on the relationships between glycans and their functions.
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DOI
高影响因子
MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Chao-Ping Hou,Ke Huang,Qiang Wang,Jin-Lan Yang,Hui-Bing Chen,Ying-Xia Zhang,Qi Liu,Zhi-Guang Li,Hui-Qing Li,Xu-Feng Wang
Tetrodotoxin (TTX), one of the most potent neurotoxins, poses a significant threat to consumer health. This study presents an advanced analytical method combining ion-paired reagent-assisted dispersive solid-phase extraction (ion-paired-dSPE) with hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS), for the quantification of TTX residues in puffer fish. Edible tissue samples were extracted twice using a 1 % formic acid–methanol solution and purified via optimized ion-paired-dSPE procedures. TTX adsorbed onto the magnetic nano-adsorbents was subsequently eluted with an acetonitrile–water–formic acid solution (60:40:1, v/v/v). Detection was achieved through electrospray ionization in positive mode, with quantification performed in multiple reaction monitoring (MRM) mode. Correlation coefficients ( r ) in matrix-matched external standard curves ranged from 0.9995 to 0.9997. The limits of detection (LOD) and quantification (LOQ) in puffer fish tissues were 0.1 μg/kg and 0.3 μg/kg, respectively. Spiked recoveries at three concentrations in negative tissue samples ranged from 73.4 % to 87.5 %, with precision under 10.6 %. The method was validated in accordance with 2002/657/EC guidelines and successfully applied to TTX detection in cultured puffer fish samples.
研究领域:
食品科学
·
分析化学
·
毒理学
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产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）