An analytical method was established for the determination of 24 food safety risk factors(18 heterocyclic aromatic amines and six mycotoxins)in synthetic meat and synthetic protein samples,and four microcystins in mic...An analytical method was established for the determination of 24 food safety risk factors(18 heterocyclic aromatic amines and six mycotoxins)in synthetic meat and synthetic protein samples,and four microcystins in microalgae samples using ultra-high-performance liquid chromatographyquadrupole/Orbitrap high-resolution mass spectrometry(UPLC-Q/Orbitrap HRMS).The novel synthetic food was extracted using 50%methanol solution and purified with 3 mL of n-hexane saturated with 50%(volume fraction)methanol solution.For the microalgae samples,extraction was performed with 80%(volume fraction)acetonitrile solution and purified using a PRiME HLB solid-phase extraction column.An ACQUITY HSS T3 column(100 mm×2.1 mm,1.8μm)was used for analyte separation,with 5 mmol/L ammonium acetate containing 0.01%(volume fraction)formic acid and methanol selected as the mobile phase.Gradient elution was adopted.Data acquisition was conducted in both positive and negative modes in full-scan data-dependent secondary scan mode(full MS-ddMS2).Matrix-matched standards were employed for quantification in the external standard method.Additionally,mzVault and TraceFinder were used to establish a screening database for 28 food safety risk factors.The results showed that the relative deviation of the exact mass of 28 food safety risk factors was less than 5×10^(-6),with good linearity in the range of 1-100μg/kg,and a correlation coefficient(r^(2))of≥0.9978.The detection limit ranged from 0.5 to 4μg/kg,and the quantification limit ranged from 1 to 10μg/kg.The method recovery ranged from 70.0%to 119.6%with relative standard deviations ranging from 0.4%to 14.6%(n=6).The method is simple,sensitive,accurate,and suitable for rapid screening of 28 food safety risk factors in novel foods.展开更多
目的利用高分辨质谱鉴定银丹心脑通软胶囊的化学成分,与网络药理学研究结合,探讨银丹心脑通软胶囊“心脑同治”的活性成分及其潜在分子作用机制。方法基于UHPLC Q-Exactive Plus Orbitrap HRMS鉴定出的银丹心脑通软胶囊化学成分;通过TC...目的利用高分辨质谱鉴定银丹心脑通软胶囊的化学成分,与网络药理学研究结合,探讨银丹心脑通软胶囊“心脑同治”的活性成分及其潜在分子作用机制。方法基于UHPLC Q-Exactive Plus Orbitrap HRMS鉴定出的银丹心脑通软胶囊化学成分;通过TCMSP和SwissADME数据库,以口服利用度≥30%、类药性≥0.18和胃肠道吸收得分为“High”、类药性至少通过3个“Yes”来筛选活性成分,再检索活性成分的潜在作用靶点;通过OMIM、DisGeNET、GeneCards、TTD和PharmGKB数据库获取与心脑血管疾病相关靶基因。使用Venny软件获得两者的交集靶点,采用Cytoscape3.9.1分析软件结合STRING数据库对交集靶点进行蛋白互作网络分析和“药物-活性成分-靶点-疾病”网络构建,确定核心成分与靶点。利用DAVID数据库对银丹心脑通软胶囊治疗心脑血管疾病的潜在靶基因进行基因本体(GO)分析及京都基因与基因组百科全书富集分析(KEGG)预测其作用机制。最后将度值排名前5的靶点与主要活性成分进行分子对接模拟以验证网络药理学结果。结果通过UHPLC Q-Exactive Plus Orbitrap HRMS技术鉴定出银丹心脑通软胶囊中82个化学成分,主要包括黄酮类、有机酸类、菲醌类和二萜内酯类化合物;经TCMSP、SwissADME数据库筛选得到活性成分34个,如槲皮素、异樱花亭、二氢丹参酮I等,相对应的靶点389个,进一步筛选出治疗心脑血管疾病相关的潜在靶基因249个。GO和KEGG富集分析显示,银丹心脑通软胶囊的活性成分可能主要通过脂质与动脉粥样硬化通路、流体剪切应力与动脉粥样硬化、AGE-RAGE、IL-17、PI3K-Akt、HIF-1、TNF等通路来发挥治疗心脑血管疾病的作用。分子对接证实STAT3、HSP90AA1等与二氢丹参酮I、山柰酚、木犀草素等具有较高的亲和力。结论本研究初步预测了银丹心脑通软胶囊“心脑同治”其作用机制与动脉粥样硬化、炎症因子、细胞凋亡和氧化应激等方面有关,为后续作用机制的深度分析及临床应用提供科学依据。展开更多
基金supported by the National Key Research and Development Program for Young Scientists of China(No.2022YFF1102900).
文摘An analytical method was established for the determination of 24 food safety risk factors(18 heterocyclic aromatic amines and six mycotoxins)in synthetic meat and synthetic protein samples,and four microcystins in microalgae samples using ultra-high-performance liquid chromatographyquadrupole/Orbitrap high-resolution mass spectrometry(UPLC-Q/Orbitrap HRMS).The novel synthetic food was extracted using 50%methanol solution and purified with 3 mL of n-hexane saturated with 50%(volume fraction)methanol solution.For the microalgae samples,extraction was performed with 80%(volume fraction)acetonitrile solution and purified using a PRiME HLB solid-phase extraction column.An ACQUITY HSS T3 column(100 mm×2.1 mm,1.8μm)was used for analyte separation,with 5 mmol/L ammonium acetate containing 0.01%(volume fraction)formic acid and methanol selected as the mobile phase.Gradient elution was adopted.Data acquisition was conducted in both positive and negative modes in full-scan data-dependent secondary scan mode(full MS-ddMS2).Matrix-matched standards were employed for quantification in the external standard method.Additionally,mzVault and TraceFinder were used to establish a screening database for 28 food safety risk factors.The results showed that the relative deviation of the exact mass of 28 food safety risk factors was less than 5×10^(-6),with good linearity in the range of 1-100μg/kg,and a correlation coefficient(r^(2))of≥0.9978.The detection limit ranged from 0.5 to 4μg/kg,and the quantification limit ranged from 1 to 10μg/kg.The method recovery ranged from 70.0%to 119.6%with relative standard deviations ranging from 0.4%to 14.6%(n=6).The method is simple,sensitive,accurate,and suitable for rapid screening of 28 food safety risk factors in novel foods.
文摘目的利用高分辨质谱鉴定银丹心脑通软胶囊的化学成分,与网络药理学研究结合,探讨银丹心脑通软胶囊“心脑同治”的活性成分及其潜在分子作用机制。方法基于UHPLC Q-Exactive Plus Orbitrap HRMS鉴定出的银丹心脑通软胶囊化学成分;通过TCMSP和SwissADME数据库,以口服利用度≥30%、类药性≥0.18和胃肠道吸收得分为“High”、类药性至少通过3个“Yes”来筛选活性成分,再检索活性成分的潜在作用靶点;通过OMIM、DisGeNET、GeneCards、TTD和PharmGKB数据库获取与心脑血管疾病相关靶基因。使用Venny软件获得两者的交集靶点,采用Cytoscape3.9.1分析软件结合STRING数据库对交集靶点进行蛋白互作网络分析和“药物-活性成分-靶点-疾病”网络构建,确定核心成分与靶点。利用DAVID数据库对银丹心脑通软胶囊治疗心脑血管疾病的潜在靶基因进行基因本体(GO)分析及京都基因与基因组百科全书富集分析(KEGG)预测其作用机制。最后将度值排名前5的靶点与主要活性成分进行分子对接模拟以验证网络药理学结果。结果通过UHPLC Q-Exactive Plus Orbitrap HRMS技术鉴定出银丹心脑通软胶囊中82个化学成分,主要包括黄酮类、有机酸类、菲醌类和二萜内酯类化合物;经TCMSP、SwissADME数据库筛选得到活性成分34个,如槲皮素、异樱花亭、二氢丹参酮I等,相对应的靶点389个,进一步筛选出治疗心脑血管疾病相关的潜在靶基因249个。GO和KEGG富集分析显示,银丹心脑通软胶囊的活性成分可能主要通过脂质与动脉粥样硬化通路、流体剪切应力与动脉粥样硬化、AGE-RAGE、IL-17、PI3K-Akt、HIF-1、TNF等通路来发挥治疗心脑血管疾病的作用。分子对接证实STAT3、HSP90AA1等与二氢丹参酮I、山柰酚、木犀草素等具有较高的亲和力。结论本研究初步预测了银丹心脑通软胶囊“心脑同治”其作用机制与动脉粥样硬化、炎症因子、细胞凋亡和氧化应激等方面有关,为后续作用机制的深度分析及临床应用提供科学依据。
