Spectrophotometric method for quantifying total lipid content and application thereof
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A spectrophotometry, lipid content technology, applied in the field of instrument analysis, can solve the problems of lack of colorimetric reagents, immaturity, inaccurate colorimetric results, etc., to achieve accurate analysis results, overcome bottlenecks, and avoid solvent interference.
Active Publication Date: 2019-07-02
TIANJIN AGRICULTURE COLLEGE
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Problems solved by technology
Although research on the quantification of total lipids by spectrophotometry has been carried out, the accuracy of quantitative results is limited by colorimetric reagents, which usually cannot completely react with lipids, resulting in inaccurate colorimetric results
Therefore, the traditional spectrophotometric technology is still immature for the quantification of total lipid content, and the biggest limitation is the lack of available colorimetric reagents
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Embodiment 1
[0016] One of the establishment of linear relationship: use petroleum ether at 60-90°C as solvent, prepare tetra-sec-octyloxyphthalocyanine nickel solution by weighing, dissolving and diluting, the concentration is 1.2×10 -5 mol L -1 . Prepare a series of solutions of peanut oil 1:0, 1:1, 1:3, 1:7, 1:15, 1:31, 1:63 (V 花生油 :V 石油醚 ), the corresponding mass volume concentrations are 0.9216, 0.4608, 0.2304, 0.1152, 0.0576, 0.0288, 0.0144g·mL -1 . The volume of the detection solution in the cuvette is constant at 3mL. Use a pipette to add 2.5mL of the above-mentioned phthalocyanine solution to the cuvette, then add 0.5mL of petroleum ether, and cover the cuvette mouth with a small piece of plastic wrap. Press the mouth of the dish with the thumb, and press the bottom of the dish with the index finger, and vibrate 3 times, as a test reference sample. Using a pipette, add 2.5mL of the above-mentioned phthalocyanine solution to the cuvette, then add 0.5mL of the peanut oil soluti...
Embodiment 2
[0019] The second establishment of linear relationship: operate as in Example 1, if the concentration of the phthalocyanine solution is prepared as 2.4 × 10 - 5 mol L -1 , after mixing with the oil sample, its concentration in the test solution was diluted to 2.0×10 -5 mol L -1 , other operations are the same as in Example 1, and the result can be equation (4).
[0020] ΔA=4.92511c l +0.01478 (4)
Embodiment 3
[0022] The third establishment of linear relationship: operate as in Example 1, if the concentration of the phthalocyanine solution is prepared as 4.8 × 10 - 5 mol L -1 , after mixing with the oil sample, its concentration in the test solution was diluted to 4.0×10 -5 mol L -1 , other operations are the same as in Example 1, and the result can be equation (5).
[0023] ΔA=6.38683c l +0.02763 (5)
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Abstract
The invention discloses a spectrophotometric method for quantifying the total lipid content and the application thereof. According to the depolymerization mechanism of lipid on a phthalocyanine aggregate, tetra-sec-octyloxy nickel phthalocyanine is used as a colorimetric material, and petroleum ether is used as a solvent to accurately quantify the total lipid content. The method comprises the steps that (1) according to standard curve preparing, a phthalocyanine solution of specific concentration is prepared; squeezed peanut oil is used as the standard lipid, and an oil-free solution is used as a reference; analyzing is carried out at 675 nm; coordinate points between the absorbance and the peanut oil concentration are fitted to acquire a linear equation; (2) according to lipid extraction,a sample to be analyzed is weighed, extracted, distilled and weighed, and lipid is recovered; and (3) according to total lipid quantifying, the phthalocyanine solution of 2.5 mL is taken and mixed with the recovered lipid sample of 0.5 mL; a lipid-free sample solution is used as a reference; analyzing is carried out at a characteristic wavelength; the absorption of the lipid sample of the same volume is removed; substituting into a linear equation is carried out; the lipid concentration, the corresponding lipid quality and the total lipid content are calculated. Compared with a traditional extraction / weighing method, the method is more accurate and can avoid the influence of a residual solvent in the recovered sample.
Description
technical field [0001] The invention belongs to the field of instrument analysis, and relates to a spectrophotometric analysis method for the total lipid content of a sample. The phthalocyanine colorimetric material in the method has stable properties, characteristic absorption in the visible spectrum, and is generally not easily disturbed by an analysis system. The experimental operation is simple and easy, the linear standard curve and linear equation are easy to obtain, and the calculation is convenient. The analysis result is more accurate. Compared with the simple weight analysis method, it can effectively avoid the interference of the residual solvent in the lipid sample. Background technique [0002] Lipid is an energy storage molecule widely distributed in biological tissue cells and is one of the structural components of biological cells. In the food industry, lipid content is an important nutritional indicator of food. In biochemical research and environmental pr...
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