High-performance liquid chromatography method and application for simultaneous determination of various taste nucleotides in fresh meat
A high-performance liquid chromatography and nucleotide technology, applied in measuring devices, instruments, scientific instruments, etc., can solve the lack of nutrition and flavor evaluation standards for chilled meat, lack of product brands and corporate brands, and doubts about the flavor of chilled meat and other problems, to achieve the effect of fast and accurate detection method, simplify the detection procedure and save the detection cost
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Embodiment 1
[0062] The establishment of embodiment 1 detection methodology
[0063] First, determine the detection conditions of high performance liquid chromatography as follows:
[0064] Columns for HPLC: ZORBAX SB-C 18 (4.6×250mm, 5μm);
[0065] Mobile phase: mobile phase A: methanol and mobile phase B: 0.04mol / L PB buffer;
[0066] The elution method adopts gradient elution, and the gradient elution program is shown in Table 1;
[0067] Table 1 Mobile phase gradient table
[0068]
[0069] Detection wavelength: 254nm;
[0070] Column temperature: 30°C;
[0071] Mobile phase flow rate: 1mL / min;
[0072] Injection volume: 10 μL.
[0073] Under the above-mentioned chromatographic conditions, the six adenosine (I), inosinic acid (IMP), hypoxanthine (Hx), adenosine monophosphate (AMP), adenosine diphosphate (ADP), and adenosine triphosphate (ATP) The standard sample is separated and analyzed to obtain the chromatographic peak figure of the standard sample under the chromatograph...
Embodiment 2
[0075] Embodiment 2 Precision experiment
[0076] The same chicken breast tissue sample was taken in 5 parts, and the retention time and content reproducibility of inosinic acid, hypoxanthine, and inosine were analyzed. The detection method included the following steps:
[0077] (1) Sample pretreatment: Grind and mix with a meat grinder, weigh 1g each, add 4mL 0.6mol / L perchloric acid into a 15mL plastic centrifuge tube, shake evenly with an oscillator, and shake the homogeneous Place the slurry in a sonicator for 10 minutes, then centrifuge at a speed of 10,000 r / min for 10 minutes, transfer 4 mL of the supernatant to a new 15 mL centrifuge tube; add 2 mL of 0.6 mol / L perchloric acid to the lower sediment, repeat In the previous step, oscillate evenly, sonicate for 10min, centrifuge at 10000r / min for 10min, transfer supernatant 2mL, combine the two supernatants; add 2.5mL 0.6mol / L Na 2 CO 3 Make the pH close to 6.0, then add 1.5mL of LPB buffer to mix, and filter with a 0.4...
Embodiment 3
[0088] Embodiment 3 recovery rate experiment
[0089] Adopt the same sample pretreatment method as in Example 2 to prepare the sample extract, take two parts of 1mL of the test solution from the same sample extract, one of which is fixed to 10mL with 5% methanol aqueous solution, and the other part is added to 2.5 mL of 0.24 mg / mL inosinic acid, 0.012 mg / mL hypoxanthine and 0.12 mg / mL inosine standard solution, then dilute to 10 mL with 5% aqueous methanol. Finally, the solution before and after adding the standard was analyzed and detected by high performance liquid chromatography. The content of each flavor nucleotide was calculated according to the formula (3) in Example 2, and the recovery rate of each component was obtained, and the parallel determination was performed 6 times. The results are shown in Table 3.
[0090] Table 3 The recovery rate experiment of several flavor nucleotides
[0091]
[0092] The experimental results show that under the chromatographic con...
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