Method for rapidly identifying and detecting tropomyosin based on intermediate infrared spectrum
A tropomyosin, infrared spectroscopy technology, applied in the field of rapid identification of allergen myosin based on mid-infrared spectroscopy, rapid identification and detection of allergen myosin in crustaceans and shellfish, to reduce The effect of cost and operation difficulty, strong universality, and simple processing method
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Embodiment 1
[0041] The aquatic product samples of Penaeus vannamei were processed to extract tropomyosin.
[0042] The pretreatment method for aquatic products is specifically to remove the shells and heads of the shrimp and remove the shrimp line, extract the shrimp meat with acetone until it is colorless, and then add 20mM Tris-HCl buffer solution (containing 0.5mol / L NaCl, pH 9.2), homogenate for 30s to obtain muscle homogenate; centrifuge at 10000r / min low temperature for 20min to obtain crude tropomyosin extract.
[0043] The tropomyosin in the crude tropomyosin extract is enriched by conventional experimental methods such as ammonium sulfate precipitation and isoelectric point precipitation, and the allergen is purified by conventional desalting means, namely dialysis, and then purified The final tropomyosin was freeze-dried to obtain tropomyosin (Tm) freeze-dried powder.
[0044] Using Spectrum GX Fourier Transform Infrared Spectrometer (Perkin Elmer Company), and solid measuremen...
Embodiment 2
[0052] Get Penaeus vannamei, grass prawns, base prawns, prawns, pipi shrimps, stone crabs, swimming crabs, clams and razor clams respectively, use the method described in Example 1 to extract its tropomyosin freeze-dried powder, and use the method of Example 1 Perform mid-infrared spectroscopy data collection.
[0053] Infrared absorption spectra of tropomyosin in different shrimp, shellfish and crab aquatic products (1800-1000cm -1 )Such as image 3 and Figure 5 shown.
[0054] Depend on image 3 and Figure 4 It can be seen that the amide I bands of the tropomyosin maps in each aquatic product are all at 1638-1647cm -1 Within the range; the amide II bands are all at 1536-1543cm -1 within range.
[0055] Depend on Figure 5 and Figure 6 It can be seen that the amide Ⅰ band and amide Ⅱ band of the tropomyosin map of stone crab and portunus crab are also within this range.
[0056] The second-order derivative is further carried out on the obtained infrared spectrum,...
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