黑米水提物功能活性成分分析

Analysis of the Functional Active Ingredients in the Water Extract from Black Rice

[目的]建立基于UHPLC-QE-MS非靶向代谢组学方法分析黑米水提物的功能活性成分。[方法]采用Waters ACQUITY UPLC BEH Amide(2.1 mm×100 mm,1.7μm)液相色谱柱;液相色谱A相为水相,含25 mmol/L乙酸铵和25 mmol/L氨水,B相为乙腈;梯度洗脱,流速为0.5 m L/min。Thermo Q Exactive HFX质谱仪在采集软件(Xcalibur,Thermo)的控制下以信息依赖模式采集质谱/质谱数据,并持续评估全扫描质谱。ESI离子源,正负离子模式下采集数据;利用一级质谱化合物的相对分子质量、分子式、二级质谱碎片离子信息等,并结合相关数据库、文献、对照品的信息等对其化学成分进行鉴定。[结果]从黑米水提物中鉴定出539种可信度很高(阈值>0.9)的化学成分,包括3种倍半萜类、3种维生素类、6种甾体类、7种有机酸类、19种黄酮类、41种生物碱类、109种氨基酸或多肽(包括8种必需氨基酸)和其他类化合物。部分化合物具有抗氧化、抗炎、抗癌、抗衰老及预防动脉硬化、降血糖和控制血压及减少患心脑血管疾病发生等功能。[结论]UHPLC-QE-MS非靶向代谢组学方法可以准确快速地分析黑米水提物的功能化学成分,为黑米的功能食品药效物质基础,质量控制以及临床应用奠定基础。

[Objective]To determine the active ingredients in the water extract from black rice with non-targeted metabolomics based on UHPLC-QE-MS.[Methods]LC-MS/MS analyses were performed using an UHPLC system(Vanquish,Thermo Fisher Scientific)with a UPLC BEH Amide column(2.1 mm×100 mm,1.7μm)coupled to Q Exactive HFX mass spectrometer(Orbitrap MS,Thermo).The mobile phase consisted of 25 mmol/L ammonium acetate and 25 ammonia hydroxide in water(A)and acetonitrile(B).The analysis was carried with elution gradient;the QE HFX mass spectrometer was used for its ability to acquire MS/MS spectra on information-dependent acquisition(IDA)mode in the control of the acquisition software(Xcalibur,Thermo).ESI ion source was used.Data were collected in both positive and negative ion mode.The relative molecular mass,molecular formula,and fragment ion information of the first-level mass spectrometry compound were used to identify their chemical constituents via combining information from related databases,literatures,and reference materials.[Results]Up to 539 chemical compositions with high reliability(cut-off value>0.9)were identified in the water extract from black rice,including 3 sesquiterpenoids,3 vitamins,6 steroids,7 organic acids,19 flavonoids,41 alkaloids,109 amino acids or polypeptides(containing 8 essential amino acids)and other compounds.Some compounds have the functions of anti-oxidation,anti-inflammation,anti-cancer,anti-aging,preventing arteriosclerosis,lowering blood sugar,controlling blood pressure and reducing the occurrence of cardiovascular and cerebrovascular diseases.[Conclusion]Non-targeted metabolomics based on UHPLC-QE-MS can be performed to effectively separate the chemical components of black rice,laying the foundation for therapeutic material basis as functional food,quality control and clinical application of black rice.