Multichannel mass spectrum derivative reagent for detecting sphingosine glucoside and sphingosine galactoside as well as preparation method and application of multichannel mass spectrum derivative reagent
A technology of sphingosine galactoside and sphingosine, which can be used in measurement devices, instruments, scientific instruments, etc., can solve the problems of high detection limit, low analytical throughput, poor versatility and the like
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Embodiment 1
[0044] Chromatographic separation and qualitative and quantitative analysis of GlcS and GalS spiked samples in delipidated plasma:
[0045] GlcS and GalS were prepared in acetonitrile to obtain 1×10 -6 Mo / L standard stock solution, and then prepared with fat-free plasma to obtain concentrations of 0.02 / 0.02, 0.2 / 0.2, 2.0 / 2.0, 20.0 / 20.0, 50.0 / 50.0, 100.0 / 100.0, 400.0 / 400.0, 800.0 / 800.0 The concentration range of the mixed standard solution of GlcS and GalS in nmol / L covers the concentration range of GlcS / GalS in the plasma of patients with Gaucher disease or Krabbe disease. CH 3 / CH 2 D / CHD 2 / 13 cd 3 / CD 3 / C 2 h 5 / C 2 h 3 D. 2 / C 2 h 2 D. 3 / C 2 D.5 -ALSF was dissolved in acetonitrile to give 2.5×10 -8 mol / L derivatization reagent acetonitrile solution; take 50 μL mixed standard solution from 8 parts of delipidated plasma mixed standard solution and a single part of acetonitrile mixed standard solution in a 1.5 mL centrifuge tube, add 300 μL H 3 BO 3 -Na ...
Embodiment 2
[0050] The detection and analysis of GlcS and GalS in the serum of a simulated Gaucher patient includes the following steps:
[0051] Take 8 healthy human serum samples from a local hospital, mix them evenly in a centrifuge tube, and centrifuge quickly at 4°C, take 500 μL of serum samples and add 500 μL methanol to a 1.5 mL centrifuge tube, vortex for 1.5 minutes and centrifuge to remove For protein precipitation, take 40 μL and place it in a 1.5 mL centrifuge tube, prepare 8 copies, and then add 10 μL of GlcS diluted with acetonitrile at a concentration of 25.0, 50.0, 100.0, 300.0, 500.0, 1000.0, 4000.0 nmol / L to each of the seven copies Standard solution, the concentration range of the solution after adding the standard covers the range of GlcS content in the serum of patients with Gaucher disease. Add 10 μL of acetonitrile solution to the remaining portion as a control. In addition, take 50 μL of the mixed standard acetonitrile solution (from a concentration of 1×10 -8 mo...
Embodiment 3
[0053] The detection of GlcS and GalS in the plasma of a simulated Gaucher patient includes the following steps:
[0054] Take 8 healthy human plasma samples from a local hospital and mix them evenly in test tubes containing ethylenediaminetetraacetic acid (EDTA), and centrifuge them quickly at 4°C. Take 2 mL of plasma samples and add 8 mL of methanol into a 15.0 mL centrifuge tube. Vortex for 2 minutes and centrifuge to remove protein precipitates. Take 40 μL and put it in a 1.5 mL centrifuge tube. Prepare 8 copies, and then add 10 μL of 25.0, 50.0, 100.0, 300.0, 500.0, 1000.0 to seven of them. 4000.0nmol / L acetonitrile diluted GlcS standard solution, the concentration range of the solution after adding the standard covers the range of GlcS content in the plasma of patients with Gaucher disease, and the remaining part is added with 10 μL acetonitrile solution as a control. In addition, take 50 μL of mixed standard acetonitrile solution (from a concentration of 1 × 10 -8 mol / ...
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