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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

Active Publication Date: 2019-12-20
河北省食品检验研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is currently no in-flight mass spectrometry database for food microorganisms, and there is a lack of positive reference samples, especially the comprehensive reference system for ready-to-use mass spectrometry identification. Standard reference samples have not yet been localized. Influencing factors

Method used

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  • Preparation method of flight time mass spectrum ready-to-use salmonella Stanley qualitative standard sample
  • Preparation method of flight time mass spectrum ready-to-use salmonella Stanley qualitative standard sample
  • Preparation method of flight time mass spectrum ready-to-use salmonella Stanley qualitative standard sample

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses a preparation method of a flight time mass spectrum ready-to-use salmonella Stanley qualitative standard sample. The preparation method comprises the following steps of adopting a specified freeze-dried matrix with the following configuration: 1640 bacteria culture medium, adding 3-7%w / w of saccharose and 6-10% of w / w of glucan. The prepared standard frozen-dried product prepared through the preparation method disclosed by the invention is excellent in performance indexes through qualitative inspection, uniformity inspection, transportation stability inspection, storagecondition stability inspection, and flight mass spectrum stability inspection, the blank of our country on this technology is filled, and the domestication of a standard reference sample of the ready-to-use spectrum identification comprehensive reference system is realized, and a flight mass spectrum database is further established for the food microorganism, thereby promoting the domestic food microorganism spectrum detection to realize qualitative development.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a method for preparing a ready-to-use qualitative standard sample of Salmonella Stanley by time-of-flight mass spectrometry. Background technique [0002] At present, the common detection methods of Salmonella in food generally use traditional routine culture physiological and biochemical detection methods, which have many disadvantages. The traditional routine culture physiological and biochemical method uses the morphology and routine physiological and biochemical characteristics of microorganisms to detect, usually including the evaluation of microbial morphology and culture characteristics. It has no special equipment requirements, and the method is classic and reliable, with the advantages of accuracy, intuition, good stability and low false positive. However, the detection cycle is long, time-consuming and labor-intensive, and there is a missed detection rate,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/62G01N1/28G01N1/42
CPCG01N27/62G01N1/28G01N1/42
Inventor 周巍张雅伦史国华张岩张翠侠张涛崔生辉
Owner 河北省食品检验研究院
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