Hao Gang, Yang Lan, Mazsaroff Istvan, Lin Melanie
Altus Pharmaceuticals, Cambridge, Massachusetts 02139, USA.
J Am Soc Mass Spectrom. 2007 Sep;18(9):1579-81. doi: 10.1016/j.jasms.2007.05.019. Epub 2007 Jun 13.
We developed a novel in vitro lipase assay based on the quantitation of fatty acids by liquid chromatography-mass spectrometry. Oleic acids enzymatically released from triolein substrates were isolated from the reaction mixture by reverse-phase chromatography, ionized in negative mode electrospray mass spectrometry and quantitated with the aid of [(13)C]-oleic acid internal standard. The enzymatic activity was measured by monitoring oleic acid productions at multiple time points. This method overcomes the substrate and pH limitations of conventional techniques and thus serves as a generic lipase activity assay.
我们开发了一种基于液相色谱 - 质谱法对脂肪酸进行定量的新型体外脂肪酶测定方法。通过反相色谱从反应混合物中分离出从三油酸甘油酯底物酶促释放的油酸，在负模式电喷雾质谱中进行离子化，并借助[（13）C] - 油酸内标进行定量。通过在多个时间点监测油酸生成来测量酶活性。该方法克服了传统技术的底物和pH限制，因此可作为一种通用的脂肪酶活性测定方法。
