Preparation method of flight time mass spectrum ready-to-use salmonella Stanley qualitative standard sample
A technology of time-of-flight mass spectrometry and standard samples, which is applied in the field of biological detection, can solve the problems of lack of positive reference samples and achieve good performance indicators
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Embodiment 1
[0032] Example 1. Screening of freeze-dried matrix.
[0033] The experimental screening of Salmonella cryopreservation conditions is mainly to confirm the composition of the freeze-dried matrix and the concentration relationship between the various substances of the freeze-dried matrix, so as to improve the survival rate and stability of the strain.
[0034] Because the previous freeze-dried matrix was milk powder, and the milk powder itself would affect the identification of Salmonella in flight mass spectrometry identification, the freeze-dried matrix was re-screened.
[0035] After research and development, 1640 cell culture medium was finally selected. Through the screening of the concentration relationship among the various substances in the dry matrix, the freeze-dried matrix was finally determined to be: 1640 cell culture medium, 5% sucrose (molecular weight: 342), and 8% dextran (molecular weight: 40,000).
Embodiment 2
[0036] Embodiment 2, preliminary preparation.
[0037] The first-generation fresh culture of Salmonella was streak-inoculated on the PCA plate, and cultured at 36°C for 24h. Use a sterile cotton swab to scrape the bacterial lawn from the plate and suspend it in the freeze-dried protective agent. Use an ultraviolet spectrophotometer to adjust the bacterial concentration, and use liquid nitrogen in a vial to prepare a standard sample.
Embodiment 3
[0038] Embodiment 3, the screening of cryopreservation process.
[0039] The prepared standard samples are placed in a vacuum freeze dryer and screened under different freezing conditions to obtain dry, ready-to-use standard samples with good vacuum tightness.
[0040] Through the screening of different freezing conditions, the freeze-drying conditions were finally determined as: pre-freezing program -20°C 200Min, -40°C 200Min; freeze-drying program -35°C 360Min, Vac 130mTorr; -30°C 360Min, Vac 130mTorr; ℃480Min, Vac 130mTorr; -20℃480Min, Vac 130mTorr; -10℃480Min, Vac 130mTorr; finally maintain at -20℃, Vac 100mTorr.
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