Reversed phase ion pair HPLC (high performance liquid chromatography) method for rapid trace detection of TTX (tetrodoxin) in fresh puffer fish blood

Abstract

The invention which belongs to the technical field of the puffer fish toxin detection concretely relates to a reversed phase ion pair HPLC method for the rapid trace detection of TTX in the fresh puffer fish blood. The method comprises the following steps: 1, acquiring and pretreating fresh puffer fish blood samples; 2, preparing a mobile phase; 3, preparing standard samples; 4, setting chromatography conditions; and 5, carrying out sample introduction detection analysis and result determination. Compared with current national standard methods of the TTX detection, the method of the inventionhas the characteristics of simple sampling, simple sample preparation, time saving and rapidness, low detection cost, high trace, qualitative and quantitative accuracy, good reappearance, no need of mass spectrums or other expensive devices, and easy popularization and application to base detection centers, provides a new detection means and a scientific basis for the safe fresh puffer eating, and performs an important effect to the promotion of the legal puffer fish eating in China and the development of the puffer fish industry and the puffer fish diet culture.

Description

technical field [0001] The invention belongs to the technical field of puffer fish puffer toxin detection technology, and in particular relates to the application field of fresh puffer toxin micro-quantity rapid and high-efficiency detection technology and puffer fish safe eating technology. Background technique [0002] The puffer fish eaten in my country now belong to Takifugu, a genus of Tetrodantidae. There are more than 20 species of this genus in the world, most of which are distributed in the coastal waters of China and Japan. It has the reputation of "the king of fish". It is popular among consumers in coastal cities of my country, Japan, South Korea and other places, and has high economic value. It has become an important artificial breeding object in my country's coastal areas. However, because this fish contains "tetrodoxin TTX", under the temptation of its delicious taste, eating puffer fish is popular even in Japan, and the laws and regulations are strict. Ther...

AI Technical Summary

Problems solved by technology

Due to the detection method of liquid-mass spectrometry, mass spectrometry equipment is expensive, and the domestic penetration rate is not high, and the detection cost is very high (only 10 mg of TTX standard product is required, imported TTX is 4,500 yuan / 1mg, and 10 mg needs 45,000 yuan), which is difficult to popularize application

Enzyme-linked immunoassay has strong specificity and high detection sensitivity, but the detection process is complicated, the cycle is long (nearly 2 working days), difficult to master, the cost of the kit is high, and it is difficult to popularize and apply

Since the current detection method mainly uses puffer fish muscle and liver samples, the pretreatment process is complex, time-consuming and consumable, and the recovery rate of TTX is low, and the analysis and detection are often interfered by impurities in the samples, which can easily cause false positive results of TTX. Therefore, it still cannot meet the needs of practical applications.

[0004]Our country is rich in puffer fish resources. In order to ensure the safety of eaters, the state currently prohibits the consumption and sale of puffer fish, and only a few experimental areas have been opened in some areas. Food points, thus causing a great waste of this resource

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