Maternal amniotic fluid phenylalanine detection method for phenylketonuria (PKU) antenatal diagnosis
A technology for phenylketonuria and phenylalanine, which is applied in the field of detection of phenylalanine in maternal amniotic fluid, and can solve problems such as inability to diagnose fetuses
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Embodiment 1
[0017] Instruments and materials
[0018] Equipment: tandem quadrupole liquid mass spectrometry (LC-MS / MS), cryogenic refrigerator (-80°C), ultrasonic crusher, high-speed refrigerated centrifuge (2,5000 rpm), micropipette.
[0019] Instrument working conditions
[0020] Chromatographic conditions: tandem quadrupole liquid mass spectrometry (LC-MS / MS) 6410B (Agilent, USA), chromatographic conditions: high performance liquid chromatography HP1260 (Agilent, USA), fast analysis column: ZORBAX SB-C 18 (50mm*2.1mm I, d, 1.8μm); mobile phase A: water (containing 0.1% formic acid) 85%, B: acetonitrile 15%; flow rate: 0.2ml / min, column oven temperature 37°C.
[0021] Mass spectrometry conditions: ion source: ESI, positive ion mode; detection method: MRM; gas flow rate: 6L / min; gas temperature: 300°C; Fragmentor: 135; Skimmer: 15; CE: 10V. Quantitative analysis of ion p-phenylalanine (m / z 166-120), labeled phenylalanine (m / z 174-128).
[0022] Reagent materials: pure water, formic ac...
Embodiment 2
[0034] Instrument materials and working conditions are the same as in Example 1.
[0035] Inspection steps:
[0036] A) Collect maternal amniotic fluid in accordance with the relevant regulations stipulated in the "Law of the People's Republic of China on Maternal and Infant Health Care", and store the collected maternal amniotic fluid in a -100°C low-temperature refrigerator. Before the experiment, the samples were thawed in a 5°C incubator, or the collected maternal amniotic fluid was directly frozen to 5°C, and then ultrasonically treated in an ultrasonic cleaner for 10 minutes.
[0037] B) Take 1ml of maternal amniotic fluid after ultrasonic treatment, keep the temperature at 5°C, use a low-temperature high-speed centrifuge at 20,000 rpm / centrifuge for 15 minutes, take 500 μl of maternal amniotic fluid supernatant after centrifugation, filter it with a 0.22 μm filter membrane, and take the filtered After the liquid, the amniotic fluid filtrate to be tested was obtained. ...
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