High-flux mass spectral method for detecting multiple amino acids in human body blood plasma
A technology for the detection of human body and amino acids, applied in the field of chemical analysis and detection, can solve the problems such as the inability to meet the needs of clinical large-scale sample detection and complexity, and achieve the effects of avoiding sample loss and human error, reducing labor costs, and improving detection efficiency.
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Embodiment 1
[0020] 1. Plasma sample pretreatment steps:
[0021] In a 96-well plate, 50 μL of plasma sample to be tested, 5 μL of internal standard, 50 μL of water and 900 μL of acetonitrile were added to each well. The above mixed solution was vortexed for 3 minutes, then stood in a refrigerator at 4°C for 30 minutes, the treated solution was centrifuged at 3000r / min for 10 minutes, and the supernatant was transferred to a 96-well sample plate with a row gun. Among them, the 17 kinds of amino acids to be tested and their internal standards are shown in the following table:
[0022] serial number
abbreviation
MW
isotopic internal standard
1
met
150
D3
2
Leu
137
D3
3
Asn
132
D3
4
Ala
90.2
D4
5
Tyr
181
13 C6
6
Phe
165.3
13 C6
7
Thr...
Embodiment 2
[0037] Determination of actual samples
[0038] According to the sample pretreatment method in Example 1, 96 normal human plasma samples (provided by Beijing Aviation General Hospital) were subjected to protein precipitation and amino acid extraction, and the amino acid extract was transferred to a 96-well sample plate, according to Example 1 Samples were analyzed and data processed under optimized mass spectrometry and liquid phase conditions, and the concentration of each amino acid was calculated according to the internal standard method, 5 of which are listed in the table below. figure 2 It is the high performance liquid chromatogram of 17 kinds of amino acids in sample 1.
[0039]
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