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Analysis method of triglyceride position isomer

A triglyceride and analysis method technology, applied in the direction of analysis materials, measurement devices, material separation, etc., can solve the problems of inaccurate quantification, lack of wide application, poor accuracy of results, etc., and achieve sample reproducibility and Good stability, less sample consumption, better qualitative and quantitative accuracy

Active Publication Date: 2022-07-01
JIANGNAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although people know the benefits of UPU triglycerides on infant health, infant formula products at home and abroad also pay more and more attention to adding functional structural lipid 1,3-dioleic acid 2-palmitic acid triglyceride (OPO), but Since there is no accurate quantitative method for the positional isomers of triglycerides, the OPO content is only determined by liquid chromatography with evaporative light detector (HPLC-ELSD) method, and sn-2 is determined by gas chromatography (GC) method. The content of palmitic acid is used as an index to evaluate this index of milk powder
The results cannot be quantified accurately due to the presence of the positional isomer OOP of OPO and other triglycerides with palmitic acid linked to the sn-2 position such as OPP
The method currently applied to the separation and identification of positional isomers in human milk fat is silver ion chromatography (Ag + -HPLC) or off-line two-dimensional chromatography, but it has only been applied in the related research of two groups of positional isomers of OPO and OOP and OPP and POP, and this method has the disadvantages of long time consumption and poor reproducibility, and is not suitable for a large number of The analysis of samples is not widely used at this stage
[0005] Adopted the detection method of liquid chromatography-mass spectrometry in the related art, carry out the analysis of triglyceride positional isomer by establishing the linear relationship between the content of triglyceride positional isomer and secondary mass spectrometry, can analyze relatively The content of triglyceride positional isomers in simple vegetable oils, but this method requires a large number of standard sample preparation, detection and data calculation, the process is cumbersome, the time cost is high, and the accuracy of the results is poor

Method used

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  • Analysis method of triglyceride position isomer
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  • Analysis method of triglyceride position isomer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1: Comparison of mass spectrometry modes

[0061] Taking O-P-O standard and breast milk samples as examples, MS E , DDA and SURVEY modes were used for detection, and the concentration of the standard was 5 μg / mL.

[0062] The result is as follows:

[0063] The MS spectra of the three mass spectrometry modes were similar. However, when analyzing complex samples, SURVEY mode is better than the other two, because the MSMS signal intensity of SURVEY mode is higher than that of DDA mode, while MS E The mode cannot ensure that the fragment ion information is specific to the same precursor ion. O-P-O Standards in MS E , DDA and SURVEY mode MS MS results see figure 1 (A)-(C), MS MS results of O-P-O of breast milk samples are shown in figure 1 (D)-(F).

Embodiment 2

[0064] Example 2: Screening of Concentration Range and Mass Spectrometry Conditions in SURVEY Mode

[0065] Determination of cone voltage: Triglyceride standard concentrations were set at 2, 5 and 8 μg / mL, cone voltage was set at 15, 20, 25, 30, 35, 40, 45, 50, 55 and 60 V, collision energy Set to 25eV.

[0066] Determination of collision energies: Triglyceride standard concentrations were set to 2, 5, and 8 μg / mL, cone voltage was set to 45 V, and collision energies were set to 6, 15, 20, 22, 25, 28, 30, and 35 eV.

[0067] Determination of the linear range of the standard concentration: the concentration of the triglyceride standard was set to 0.4, 0.8, 1, 2, 4, 5, 6, 8, 10 and 12 μg / mL, the cone voltage was set to 45 V, and the collision energy was set to 25eV.

[0068] The result is as follows

[0069] The total ion current diagram of ABC-type triglyceride standard products O-P-La, O-P-S, O-P-L is shown in figure 2 (A), see the total ion chromatogram of AAB-type trigl...

Embodiment 3

[0070] Example 3: Establishment of AAB-type triglyceride standard

[0071] Sn-A-A-B and sn-A-B-A triglyceride isomers were mixed in mass ratios of 1:0, 3:1, 1:1, 1:3 and 0:1, and the concentration of the mixed standard was 5 μg / mL.

[0072] Under the condition that the cone hole voltage is 45V and the collision energy is 25eV, the SURVEY mode is selected for detection. Taking O-P-O as an example, the results of the MS spectra of the ratios of sn-O-P-O:sn-O-O-P of 1:0, 1:1 and 0:1 are shown in Figure 5 (A)-(C).

[0073] The result is as follows

[0074] Taking the ratio of sn-A-B-A in the mixture as the abscissa, [R A-A ] + / [R A-A ] + +[R A-B ] + As the ordinate, perform linear fitting to obtain the fitted curve and R 2 . see linear curve Image 6 .

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Abstract

The invention provides an analysis method of a triglyceride position isomer, belongs to the field of lipid analysis, and adopts a supercritical fluid chromatography-mass spectrometry method to carry out qualitative analysis on the triglyceride position isomer. And carrying out quantitative analysis on the triglyceride position isomer by adopting an abundance method, specifically, obtaining the abundance of fragment ions by virtue of secondary mass spectrometry, and analyzing the relative content of the triglyceride position isomer by virtue of the abundance, the triglyceride comprises ABC type triglyceride and / or AAB type triglyceride. And the purpose of accurately and quickly analyzing the content of the triglyceride position isomer can be achieved.

Description

technical field [0001] The invention relates to the field of lipid analysis, in particular to an analysis method for positional isomers of triglycerides, which is especially suitable for the qualitative and quantitative determination of positional isomers of triglycerides such as natural lipids, biological samples, complex samples and structural lipids . Background technique [0002] The structures of triglycerides vary widely among species of different biological origins in nature. In vegetable oils, saturated fatty acids are generally located at the sn-1 / 3 position, and unsaturated fatty acids are located at the sn-2 position; in milk fat, especially human milk fat, saturated fatty acids are mainly located at the sn-2 position; in addition, in different species of milk There are also differences in the positional structure of fatty acids (FAs) on fatty triglycerides. [0003] The positional distribution of fatty acid types on triglycerides has an important influence on t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/72
CPCG01N30/02G01N30/06G01N30/72Y02P20/54
Inventor 韦伟张星河金俊金青哲王兴国
Owner JIANGNAN UNIV
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