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Single-column two-dimensional liquid chromatography-mass spectrometry analysis method for triglyceride in edible oil and application of method

A technology of two-dimensional liquid chromatography and triglyceride, which is applied to the single-column two-dimensional liquid chromatography-mass spectrometry analysis of triglyceride in edible oil and its application field, which can solve the large error in quantitative analysis, cumbersome operation and high price To achieve high-throughput detection, ensure safety, and simple operation

Active Publication Date: 2015-06-17
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The advantage of the offline mode is that it is easy to operate, and the separation conditions of each dimension can be optimized independently, but the disadvantage is that the chromatographic column needs to be replaced between the two dimensions, and because the mobile phases used in the first and second dimensions are immiscible, replacement After the chromatographic column, it is necessary to repeatedly flush the pipeline with isopropanol, and then use normal phase or reverse mobile phase to balance the chromatographic column, resulting in cumbersome operation, waste of solvent and environmental pollution; while the online mode has a high degree of automation and reproducibility It has the advantages of better performance, but its device is more complicated. In addition, because the two-dimensional solvents are not miscible, there is also the problem of solvent incompatibility.
[0005] When mass spectrometry (MS) is used for the quantitative analysis of complex compounds of triglycerides, in theory any compound quantified by MS must accurately compare its own peak intensity with the peak intensity of the stable isotope internal standard compound or use a standard to make a standard curve. To achieve quantitative analysis, however, the composition of triglycerides in edible oils is very complex, and it is not feasible to use dozens or even hundreds of isotope internal standard compounds to analyze triglyceride mixtures, and because commercial TAG standards are expensive And the types are very limited, so it is impossible to use the standard curve of all triglycerides in vegetable oils to achieve quantitative analysis
The number of double bonds and acyl chain length of triglycerides will affect the sensitivity of triglycerides in MS. Therefore, only considering the chromatographic peak area and ignoring the difference in mass spectrometry response may lead to large errors in quantitative analysis.

Method used

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  • Single-column two-dimensional liquid chromatography-mass spectrometry analysis method for triglyceride in edible oil and application of method
  • Single-column two-dimensional liquid chromatography-mass spectrometry analysis method for triglyceride in edible oil and application of method
  • Single-column two-dimensional liquid chromatography-mass spectrometry analysis method for triglyceride in edible oil and application of method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1: Separation and detection of three kinds of triglycerides with the same ECN value and different numbers of double bonds in oil samples

[0069] Separation and detection of three kinds of triglycerides with the same ECN value and different numbers of double bonds in oil samples: Agilent Zorbax Eclipse Plus phenyl-hexyl (4.6mm×250mm i.d.) with both hydrophobic interaction and π-π complexation is used as Separation column, the mobile phase is 100% methanol, the flow rate is 1mL / min, the injection volume is 10μL, the ultraviolet detection wavelength is 205nm, and the column temperature is 35°C for liquid chromatography separation. figure 1 It is the liquid chromatogram of three kinds of triglycerides (PPO, OOP, SOL) with the same ECN value and different numbers of double bonds in the separated oil samples. The concentration of the samples used is 1mg / ml.

Embodiment 2

[0070] Example 2: Separation and detection of 6 kinds of triglycerides with different ECN values ​​in oil samples

[0071] Separation and detection of 6 kinds of triglycerides with different ECN values ​​in oil and fat samples: Agilent Zorbax Eclipse Plus phenyl-hexyl (4.6mm×250mm i.d.) with both hydrophobic interaction and π-π complexation was used as the separation column, mobile phase It is 100% acetonitrile, the flow rate is 1mL / min, the injection volume is 10μL, the ultraviolet detection wavelength is 205nm, and the column temperature is 35°C, for liquid chromatography separation. figure 2 It is the liquid chromatogram of 6 kinds of triglycerides (LLL, LLP, POL, SOL, OOS, OOA) with different ECN values ​​in the separated oil samples, and the concentration of the samples used is 1mg / ml.

Embodiment 3

[0072] Example 3: Simultaneous separation and detection of triglycerides with different ECN values ​​and 12 triglycerides with the same ECN value but different numbers of double bonds in oil samples

[0073] Simultaneous separation and detection of triglycerides with different ECN values ​​and triglycerides with the same ECN value but different numbers of double bonds in oil samples: the chromatographic column is Agilent Zorbax Eclipse Plus phenyl-hexyl (4.6mm×250mm i.d.), and the mobile phase is A : Acetonitrile, B: Methanol (containing 0.4% ammonia water, v / v), gradient elution: 0-7.5min, 50%A; 7.5-8.0min, 50-28%A; 8.0-12.9min, 28%A ; 12.9-13.0min, 28-50%A; 13.0-26.0, 50%A; flow rate 0.8mL / min, injection volume 10μL, UV detection wavelength 205nm, column temperature 35℃, for liquid chromatography separation. image 3 It is used to simultaneously separate triglycerides with different ECN values ​​and 12 triglycerides with the same ECN value but different numbers of double bon...

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Abstract

The invention relates to a single-column two-dimensional liquid chromatography-mass spectrometry analysis method for triglyceride in edible oil. The single-column two-dimensional liquid chromatography-mass spectrometry analysis method for the triglyceride in the edible oil is characterized by comprising the following steps: 1) a qualitative method for the triglyceride in the edible oil: (1) performing online single-column two-dimensional liquid chromatography-mass spectrometry separation, and (2) performing offline single-column two-dimensional liquid chromatography-mass spectrometry separation; 2) a quantitative method for the triglyceride in the edible oil; 3) application of the single-column two-dimensional liquid chromatography-mass spectrometry analysis method in the fidelity of the edible oil: (1) by adopting the methods of the step 1) and the step 2), extensively testing the edible oil of different brands of the same variety and doped samples to obtain liquid chromatography-mass spectrometry data of the triglyceride under the same conditions, and (2) analyzing, identifying and distinguishing the types of the edible oil and identifying the doped edible oil by combining principal components. The method is easy to operate; high-efficiency and high-throughput detection of the triglyceride can be realized.

Description

technical field [0001] The invention relates to a single-column two-dimensional liquid chromatography-mass spectrometry analysis method for triglyceride in edible oil and its application in oil identification and doping detection (in oil preservation). Background technique [0002] As people pay more and more attention to the safety and nutrition of edible oil, eating good oil and eating healthy oil has become a new demand for Chinese consumers to pay attention to their own health and ensure normal nutritional intake. However, the phenomenon of unscrupulous traders mixing fakes with real ones and shoddy ones still occurs. Effectively monitoring the quality of edible oil is an important link and means to ensure the safety of edible oil. [0003] Triacylglycerols (TAGs) are formed by the condensation of one glycerol molecule and three fatty acid molecules, accounting for more than 95% of edible oil components, and have an important impact on the physicochemical and nutritiona...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/88
Inventor 魏芳胡娜董绪燕陈洪吕昕
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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