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产品定义
短链脂肪酸(Short-chain fatty acids , SCFAs)is一组由五pcs或以下of碳原子组成of饱and脂肪酸. 短链脂肪酸generallyis由厌氧bacteria进行糖酵解制造出来of,SCFA会rapid地in胃肠道of上皮cell被吸收, is肠道cellofmainly能量来源. 本方法utilizing靶向(GC-SIM)ofanalyze技术方法进行detect, 对粪便/内容物/血液样本中of7种短链脂肪酸进行绝对定量analyze.
技术路线
技术优势
1. 高Specificity, 高Accuracy: adoptingGC-SIM精确定性, 绝对定量
2. 专属kit: 专for短链脂肪酸develop
3. 严格质控: 内标+外标双重质控, standard人工校对, 标曲R2>0.99
Detection Indicators
编号 | 代谢物名称 | Metabolites |
1 | 乙酸 | Acetic acid |
2 | 丙酸 | Propanoic acid |
3 | 丁酸 | Butyric acid |
4 | 异丁酸 | Isobutyric acid |
5 | 戊酸 | valeric acid |
6 | 异戊酸 | Isovaleric acid |
7 | 己酸 | Hexanoic acid |
Data Analysis
应用领域
1. 疾病research: 肠道炎症, 结肠癌症etc.疾病ofresearch
2. 肠道菌群相关机制research:SCFAs对代谢调节作用机制
| Sample Type | 送样量 | 预processandstore |
| 动物tissue | ≥200 mg | PBS wash去除残留血液and污染物, 冲洗干净. 用tissue剪或手术刀将tissue剪切成 1cm3 左右of小块, 样品称重, etc.量分装. 液氮速冻, -80°C store, 干冰transport. |
| Serum | ≥300 μL | 收集好of全血室温静置 30-60 min, or 4°C 放置 2h, 3000 g centrifuge 10 min, 取上清, 液氮速冻, -80°C store, 干冰transport. |
| 血浆 | ≥300 μL | 收集好of全血加入抗凝剂 (推荐adopting肝素抗凝剂, 不recommendadopting柠檬酸or EDTA) , 室温静止 30 minutes, 1300g-2000g centrifuge 10min, 取上清, 液氮速冻, -80°C store, 干冰transport. |
| 尿液 | ≥200 μL | 5000×g 4°C centrifuge 30-60min, 取上清, -80°C store, 干冰transport. |
| 粪便 | ≥300 mg | 新鲜粪便样本收集好后, 立即用液氮速冻process15 min, store至-80℃冰箱 |
| 肠道内容物 | ≥300 mg | 新鲜肠道内容物样本收集好后, 立即用液氮速冻process15 min, store至-80℃冰箱 |
| 乳汁 | ≥200 μL | 收集母乳, -80°C store, 干冰transport. |
| 唾液 | ≥500 μL | 1000g-2000g 4°C centrifuge 10min, (或using 0.22μm 滤膜过滤) , 取上清, -80°C store, 干冰transport. |
| 淋巴液, 脑脊液, 羊水, 胆汁etc.体液 | ≥200 μL | 1000g-2000g 4°C centrifuge 10min, (或using 0.22μm 滤膜过滤) , 取上清, -81°C store, 干冰transport. |
| 穿刺样品 | ≥500 mg | 收集样品, adopting PBS wash去除样品表面泥土或污染物 (务必清洗干净! ! ) , 样品称重, etc.量分装, 液氮速冻, -80°C store, 干冰transport. |
| 植物种子, 果肉 | ≥200 mg | 收集样品, 样品称重, etc.量分装, 液氮速冻, -80°C store, 干冰transport. |
| 草本植物, 藻类, 蕨类 | ≥200 mg | 收集样品, adopting PBS wash去除样品表面泥土或污染物 (务必清洗干净! ! ) , 样品称重, etc.量分装, 液氮速冻, -80°C store, 干冰transport. |
| 植物花粉 | ≥50 mg | 收集样品, 样品称重, etc.量分装, 液氮速冻, -80°C store, 干冰transport. |
| 植物体液 | ≥300 μL | 5000×g 4°C centrifuge 30-60min, 取上清, -80°C store, 干冰transport. |
| 软体动物tissue | ≥500 mg | 收集样品, adopting PBS wash去除样品表面泥土或污染物 (务必清洗干净! ! ) , 样品称重, etc.量分装, 液氮速冻, -80°C store, 干冰transport. |
| 低etc.bacteriacell | ≥10^9 | centrifuge收集etc.量菌体 (保证每copies收集of菌体cell数量一致! ! ) , adopting PBS 清洗三times, 每times清洗后 5000g 4°Ccentrifuge 5 minutes, 完全弃去上清, 收集in 1.5ml centrifuge管中. 液氮速冻, -80°C store, 干冰transport. |
| 菌体 (fungus类) | ≥500 mg | 收集菌体 (保证每copies收集of菌体数量or重量一致! ! ) , adopting PBS 清洗三times, 每times清洗后 5000g 4°C centrifuge 5 minutes, 完全弃去上清, 收集in 1.5ml centrifuge管中. 液氮速冻, -80°C store, 干冰transport. |
| 悬浮culturecell | ≥10^7 | centrifuge收集悬浮cell (保证每copies收集ofcell数量一致! ! ) , adopting PBS 清洗两遍, 然后adopting生理盐水 (0.9%氯化钠溶液) 洗一遍, 每times清洗后adopting小于 1000g centrifuge力 4°C 低速centrifuge 3 minutes, 完全弃去上清 (上述整pcs过程需尽量迅速操作) , 收集in 1.5ml centrifuge管中. 液氮速冻后-80°C store, 干冰transport. |
| 贴壁culturecell | ≥10^7 | 收集culture好of贴壁cell (保证每copies收集ofcell数量一致! ! ) 去除干净Culture Medium, adopting预冷of PBS 洗两遍, 然后再times加入预冷ofPBS, 把culture容器中of所havecell都收集下来 (一定要刮取干净! ! ) , 收集in 1.5 mL centrifuge管中, 4°C 低速centrifuge, 完全弃去上清 (上述整pcs过程需尽量迅速操作) . cell沉淀in液氮速冻后-80°Cstore, 干冰transport. |
| Cell Culture上清 | ≥10 mL | 1000 g, 4°Ccentrifuge3 minutes, 取上清, -80°C store, 干冰transport. |
Sample Preparation: 确保样品is干燥of, 因for水分可能会影响GC-MSofanalyze结果. such as果样品中含have水分, 可能need进行干燥或去除水分ofprocess.
衍生化: 短链脂肪酸usuallyis羧酸, 不易挥发, 因此needthrough衍生化反应转化for易挥发of衍生物, 以便进行GC-MSanalyze. 常用of衍生化方法including酯化, 硅烷化etc..
instrument设置: in进行GC-MSanalyze时, need选择合适of色谱柱, detect器as well as相应ofanalyze条件. for example, 选择合适of色谱柱can实现对不同短链脂肪酸of分离; 选择合适ofdetect器 (such as电子捕获detect器或质谱detect器) can提高analyzeofSensitivityandSpecificity.
Quality Control: for了确保analyze结果ofAccuracy, need进行Quality Control. 这includingusingstandard进行定量analyze, 重复determine样品as well as检查instrument状态etc..
数据解析: GC-MSanalyze会产生大量of数据, need进行仔细of数据解析. 这including对色谱峰进行识别, 计算峰面积, 确定短链脂肪酸of种类and浓度etc..
