Method for detecting carboxy methyl lysine (CML) in soybean sauce
A carboxymethyl lysine and detection method technology, which is applied in the field of food detection, can solve the problems of inability to obtain the CML content of the soy sauce system completely, there is no pre-treatment method for the detection of free CML, and the inability to completely precipitate and separate low-molecular-weight polypeptides, etc., to achieve detection The mass spectrometry response signal is good, the detection accuracy is improved, and the cost is low.
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Embodiment 1
[0053] The pre-processing of CML detection is performed on different brands of Japanese soy sauce, different brands of Chinese soy sauce and mixed soy sauce, including the following steps:
[0054] a. Pre-processing of free CML, including the following steps:
[0055] (1) Take 0.1mL soy sauce sample, dilute to 10mL with 10% methanol solution, and vortex to mix;
[0056] (2) Take 1mL of diluent and use activated C 18 Clean up the solid phase extraction column, add 5mL of 10% methanol solution to elute, and collect the eluate;
[0057] (3) Take 1 mL of the eluate and pass it through a 0.22 μm microfiltration membrane to obtain the desired sample 1 to be tested.
[0058] At the same time as the mixed soy sauce sample is detected, the impurity removal effect analysis is carried out, which specifically includes the following steps:
[0059] Using a UV-visible spectrophotometer (50Conc, VARIAN, USA), measure the absorbance value of the mixed soy sauce sample after the above pretr...
Embodiment 2
[0099] a. Pre-detection treatment of free CML, the specific steps are:
[0100] (1) Take 0.1mL soy sauce sample, dilute to 10mL with 10% methanol solution, and vortex to mix;
[0101] (2) Take 1mL of diluent and use activated C 18 Clean up the solid-phase extraction column, add 8 mL of 10% methanol solution to elute, and collect the eluate;
[0102] (3) Take 1 mL of the eluate and pass it through a 0.22 μm microfiltration membrane to obtain the desired sample 1 to be tested.
[0103] b. Pre-detection treatment of bound CML, the specific steps are:
[0104] (1) Take 7.5mg of soy sauce with equivalent peptide nitrogen content, add boric acid buffer solution (0.5mol / L, pH9.2), after adding, make the final molar concentration of sodium borate in the system reach 0.2mol / L, and add sodium borohydride Solution (2 mol / L sodium borohydride solution dissolved in 0.1 mol / L sodium hydroxide solution), after adding to make the final concentration of sodium borohydride in the system reac...
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