CN110554107B - Method for analyzing triglyceride components in edible oil by high performance liquid chromatography-mass spectrometry - Google Patents

Method for analyzing triglyceride components in edible oil by high performance liquid chromatography-mass spectrometry Download PDF

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CN110554107B
CN110554107B CN201910750059.5A CN201910750059A CN110554107B CN 110554107 B CN110554107 B CN 110554107B CN 201910750059 A CN201910750059 A CN 201910750059A CN 110554107 B CN110554107 B CN 110554107B
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edible oil
liquid chromatography
high performance
performance liquid
isopropanol
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CN110554107A (en
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许旭
陆湘婷
张世鼎
龚灿
鲁彦
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Shanghai Institute of Technology
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Abstract

The invention relates to a method for analyzing triglyceride components in edible oil by high performance liquid chromatography-mass spectrometry, which comprises the steps of dissolving an edible oil sample in an organic solvent, taking n-octane-isopropanol or n-heptane-isopropanol as a mobile phase, taking a porous graphite carbon column as a stationary phase, and adopting a high performance liquid chromatography-mass spectrometer for separation and analysis. Compared with the prior art, the invention has the advantages of low mobile phase toxicity, low ultraviolet background, simple and convenient operation, capability of separating isomers and high accuracy of separation and detection results. The method can be used for analyzing various triglyceride components in vegetable oil and animal oil, can be applied to identification of adulterated vegetable oil containing animal oil components, and is a direct and effective edible oil analysis method.

Description

Method for analyzing triglyceride components in edible oil by high performance liquid chromatography-mass spectrometry
Technical Field
The invention relates to the field of analysis and detection, in particular to a method for analyzing triglyceride components in edible oil by high performance liquid chromatography-mass spectrometry.
Background
Edible oils, including vegetable oils and animal oils, are composed primarily of various Triglyceride (TAGs) components and lower amounts of diglycerides, free Fatty Acids (FAs), phospholipids and other minor components. Of these, triglycerides, also known as Triacylglycerols (TAGs), are the most important nutrients, accounting for approximately 95% to 98%. They constitute an important part of the human diet and their uncontrolled intake may lead to coronary heart disease, dyslipidemia and obesity. On the other hand, healthy fat intake may also reduce the incidence of cardiovascular disease.
Currently, some manufacturers mix low cost edible oils into high value edible oils for profit, and gain corresponding benefits through adulteration. For example, olive oil is blended with a certain amount of soybean oil or peanut oil, corn oil is blended with animal oil, etc. In order to obtain huge profits, some lawbreakers also use the illegal cooking oil to impersonate the edible oil or mix the illegal cooking oil with the edible oil, which can bring extremely adverse threats to the health of people.
Different edible oils and adulterated edible oils can be identified by utilizing the difference of triglyceride compositions in different edible oils. This requires separate analysis of the triglycerides in the edible oil, and in particular the identification of triglyceride components with different isomers is key to the detection of adulterated edible oils.
Several analytical techniques are currently available for qualitative and quantitative triglyceride determination, ranging from classical spectroscopic methods such as infrared, raman and Nuclear Magnetic Resonance (NMR) to the more recent high temperature gas chromatography-mass spectrometry, High Performance Liquid Chromatography (HPLC), and high temperature gas-mass spectrometry (GC-MS) and liquid-mass spectrometry (LC-MS) techniques, of which HPLC and LC-MS based thereon have the potential to separate triglyceride isomers, the most studied.
HPLC-based methods are classified as normal phase, reversed phase, and silver ion stationary phase. The selectivity of the triglyceride is poor and the use amount is small. Reverse phase HPLC has become an important triglyceride separation system using a C18 or C8 stationary phase and a non-aqueous mobile phase. At present, new liquid chromatography technologies such as hydrophilic stationary phase, monolithic column, multi-column series, super-effective stationary phase, core-shell fixing and the like are used in separation research of TAGs, and people's understanding of TAG difference in different oils is rapidly improved. The reverse phase liquid chromatography is basically based on carbon number separation, and has weak separation ability for the same equivalent carbon number (ECN, the number of double bonds subtracted from the total carbon number of acyl chains) and isomers, and even if a multi-column tandem or super-effect column is adopted, satisfactory separation is often difficult to obtain. High temperature gas chromatography also has similar circumstances in the separation of TAGs.
Silver ion HPLC generates interaction with carbon-carbon double bonds through silver ions on a stationary phase, TAG can be separated according to the number of the double bonds, and the method becomes an important supplement of reversed-phase liquid chromatography. Early HPLC of silver ions was achieved by binding silver ions to a silica gel stationary phase, with short column life and insufficient stability. The appearance of the second generation silver ion stationary phase based on sulfonic acid ion exchange groups enables the rapid development of silver ion HPLC in triglyceride separation, and Dugo P and the like identify 5% of beef tallow doped in lard by using the ratio of isomers sn-POP/sn-PPO as an index through a method based on silver ion HPLC. However, the lifetime and stability of such stationary phase columns are still not ideal. The third generation silver ion stationary phase based on sulfydryl appeared in 2012 is obviously improved compared with the previous silver ion stationary phase based on sulfonic acid group, the service life after single silver is greatly prolonged, and the silver ion stationary phase can be used for preparing, separating and purifying polyene fatty acid, but the related research still has certain obstacles due to the lower column efficiency of the silver ion column and the defects of the column technology.
Disclosure of Invention
The invention aims to solve the problems and provide a method for analyzing triglyceride components in edible oil by high performance liquid chromatography-mass spectrometry, which has the advantages of capability of separating isomers, high accuracy of separation detection results, simple method and simple and convenient operation, can obviously improve the detection quality and improve the accuracy of detection effects.
The purpose of the invention is realized by the following technical scheme:
a method for analyzing triglyceride components in edible oil by high performance liquid chromatography-mass spectrometry comprises dissolving an edible oil sample in an organic solvent, taking n-octane-isopropanol or n-heptane-isopropanol as a mobile phase, taking a porous graphite carbon column as a stationary phase, and performing separation analysis by using a high performance liquid chromatography-mass spectrometer.
The organic solvent is n-hexane.
The method comprises the specific steps of dissolving an edible oil sample in n-hexane, and separating and analyzing triglyceride in the edible oil sample by using a high performance liquid chromatography-mass spectrometer, wherein the specific HPLC and MS conditions of the high performance liquid chromatography-mass spectrometer are as follows:
(1) HPLC conditions are as follows: the flow rate is 0.1-0.3 mL/min; the temperature is 50-90 ℃; the ratio of mobile phase n-octane-isopropanol or n-heptane-isopropanol is 9: 1(v/v) to 5: 5(v/v), and the ultraviolet detection wavelength is 200 nm-250 nm;
(2) MS conditions: APCI mode, positive ion mode; the mass range is 500-1100 m/z; the source temperature is 200-350 ℃; desolvation Line (DL) temperature is 150-250 ℃; the flow rate of the atomized gas is 2L/min to 7L/min; the flow rate of the drying gas is 3L/min-7L/min.
Preferably, the specific HPLC and MS conditions for the high performance liquid chromatography-mass spectrometer are as follows:
(1) HPLC conditions: the flow rate is 0.25 mL/min; the temperature is 60 ℃; mobile phase n-octane-isopropanol (7: 3); ultraviolet detection wavelength is 215 nm;
(2) MS conditions: APCI mode, positive ion mode; the mass range is 500-1100 m/z; the source temperature is 300 ℃; desolvation Line (DL) temperature 200 deg.C; the flow rate of the atomized gas is 2.5L/min; the flow rate of the drying gas was 5L/min.
The column temperature of the porous graphite carbon column is 50-90 ℃.
Dissolving an edible oil sample in n-hexane, and diluting to 0.1-1.0 mg/mL by using a mobile phase.
The high performance liquid chromatography-mass spectrometer adopts an LCMS-2020 liquid mass spectrometer.
The porous graphite carbon column adopts a Hypercarb column.
The edible oil sample comprises vegetable oil and animal oil.
Compared with the prior art, the invention has the beneficial technical effects that:
the high performance liquid chromatography and the mass spectrum combined method of the graphitized carbon chromatographic column use n-octane-isopropanol or n-heptane-isopropanol as a mobile phase, have the capability of separating isomers, have high accuracy of separation detection results, have simple method and simple and convenient operation, can obviously improve the detection quality and improve the accuracy of detection effects.
The method has the advantages of low toxicity of the mobile phase, low ultraviolet background and simple and convenient operation, can analyze various triglyceride components in the vegetable oil and the animal oil, can be applied to the identification of adulterated vegetable oil containing animal oil components, and is a direct and effective edible oil analysis method.
The problem of toluene contamination of the mobile phase when effecting the separation of the triacylglycerol acyl positional isomer, SPO/SOP, limits its widespread use. The graphitized carbon chromatographic column is adopted, a novel environment-friendly n-octane-isopropanol or n-heptane-isopropanol system is provided as a mobile phase, separation of common triglyceride acyl position isomer SPO/SOP components in the edible oil is realized, better practical application capability is realized, and the graphitized carbon chromatographic column can be used for identification of adulterated vegetable edible oil by combining research of characteristic components in the edible oil.
Drawings
FIG. 1 is a mass spectrometric separation profile of corn oil;
FIG. 2 is a graph of mass spectrometry of soybean oil;
FIG. 3 is a graph of mass spectral separation of lard oil;
FIG. 4 is a graph of mass spectrometry of corn oil spiked with lard;
corn oil A, corn oil B, lard oil 0.1%, corn oil C, lard oil 0.5%, corn oil D and lard oil 1%
FIG. 5 is a graph of mass spectrometry of soybean oil spiked with lard;
Soybean oil A, soybean oil B and lard oil 0.1%
Detailed Description
The invention is described in detail below with reference to the figures and specific embodiments.
Equipment and materials used in the examples:
the apparatus was an LCMS-2020 liquid phase mass spectrometer (Shimadzu, Japan), a Hypercarb column (100 mm. times.2.1 mm ID, particle size 5 μm) (Saimer Feishel science, USA);
the material comprises n-octane and isopropanol as mobile phase, and corn oil, soybean oil and lard as raw materials.
The specific HPLC and MS conditions for high performance liquid chromatography-mass spectrometry were as follows:
(1) HPLC conditions: the flow rate is 0.1-0.3 mL/min; the temperature is 50-90 ℃; the ratio of mobile phase n-octane-isopropanol or n-heptane-isopropanol is 9: 1(v/v) to 5: 5(v/v), and the ultraviolet detection wavelength is 200 nm-250 nm;
(2) MS conditions: APCI mode, positive ion mode; the mass range is 500-1100 m/z; the source temperature is 200-350 ℃; desolvation Line (DL) temperature is 150-250 ℃; the flow rate of the atomized gas is 2L/min to 7L/min; the flow rate of the drying gas is 3L/min-7L/min.
The column temperature of the porous graphite carbon column is 50-90 ℃.
Example 1
mu.L of corn oil (0.5mg/mL) was injected into LC-MS under the specific HPLC and MS conditions as follows: (1) HPLC conditions: the flow rate is 0.25 mL/min; the temperature is 60 ℃; mobile phase n-octane-isopropanol (7: 3); ultraviolet detection wavelength is 215 nm; (2) MS conditions: APCI mode, positive ion mode; the mass range is 500-1100 m/z; the source temperature is 300 ℃; desolvation Line (DL) temperature 200 deg.C; the flow rate of the atomized gas is 2.5L/min; the flow rate of the drying gas was 5L/min. Monitoring the separation effect of corn oil in Scan mode, as shown in fig. 1, can separate and characterize 14 kinds of triglyceride under the condition that the S/N ratio is greater than 3.
Example 2
mu.L of soybean oil (0.5mg/mL) was injected into LC-MS and the separation effect of soybean oil was monitored in Scan mode. As shown in FIG. 2, 14 triglycerides can be isolated and characterized with a signal-to-noise ratio S/N > 3.
Example 3
mu.L of lard (0.5mg/mL) was injected into LC-MS and the separation effect of lard was monitored in Scan mode. As shown in FIG. 3, 20 triglycerides can be isolated and characterized with a signal-to-noise ratio S/N > 3.
Example 4
The corn oil was mixed with 1% lard, 0.5% lard and 0.1% lard, respectively, in a sample volume of 10 μ L. Monitoring is performed in SIM mode. As a result, the signal to noise ratio of SPO contained in the corn oil is relatively small, and the signal to noise ratio of SPO in the corn oil is increased after 0.1-1% lard is added. Thus, the incorporation of lard in corn oil can be identified and the spectrum is shown in FIG. 4.
Example 5
Soybean oil was blended with 0.1% lard, and the sample size was 6. mu.L. Monitoring is performed in SIM mode. As a result, the signal to noise ratio of SPO contained in soybean oil is relatively small, and the signal to noise ratio of soybean oil doped with 0.1% lard is remarkably increased. Thus, the incorporation of lard in soybean oil can be identified, and the spectrum is shown in fig. 5.
The embodiments described above are described to facilitate an understanding and use of the invention by those skilled in the art. It will be readily apparent to those skilled in the art that various modifications to these embodiments may be made, and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above embodiments, and those skilled in the art should make improvements and modifications within the scope of the present invention based on the disclosure of the present invention.

Claims (5)

1. A method for analyzing triglyceride components in edible oil by high performance liquid chromatography-mass spectrometry is characterized in that an edible oil sample is dissolved in an organic solvent, n-octane-isopropanol or n-heptane-isopropanol is used as a mobile phase, a porous graphite carbon column is used as a stationary phase, and a high performance liquid chromatography-mass spectrometer is used for separation and analysis;
the organic solvent is n-hexane;
the porous graphite carbon column adopts a Hypercarb column, the specification is 100 mm multiplied by 2.1 mm ID, and the particle size is 5 mu m;
the method comprises the specific steps of dissolving an edible oil sample in n-hexane, and separating and analyzing triglyceride in the edible oil sample by using a high performance liquid chromatography-mass spectrometer, wherein the specific HPLC and MS conditions of the high performance liquid chromatography-mass spectrometer are as follows:
(1) HPLC conditions: the flow rate is 0.1-0.3 mL/min; the temperature is 50-70 ℃; the ratio of mobile phase n-octane-isopropanol or n-heptane-isopropanol is 9: 1-5: 5, v/v, the ultraviolet detection wavelength is 200-250 nm;
(2) MS conditions: APCI mode, positive ion mode; the mass range is 500-1100 m/z; the source temperature is 200 ℃ and 350 ℃; desolvation Line (DL) temperature 150-; the flow rate of the atomized gas is 2-7L/min; the flow rate of the drying gas is 3-7L/min.
2. The method for analyzing triglyceride composition in edible oil by high performance liquid chromatography-mass spectrometry as claimed in claim 1, wherein the specific HPLC and MS conditions of the high performance liquid chromatography-mass spectrometer are as follows:
(1) HPLC conditions are as follows: the flow rate is 0.25 mL/min; the temperature is 60 ℃; the ratio of the mobile phase n-octane to the isopropanol is 7:3, v/v; ultraviolet detection wavelength is 215 nm;
(2) MS conditions: APCI mode, positive ion mode; the mass range is 500-1100 m/z; the source temperature is 300 ℃; desolvation Line (DL) temperature 200 deg.C; the flow rate of the atomized gas is 2.5L/min; the flow rate of the drying gas was 5L/min.
3. The method for analyzing triglyceride components in edible oil by high performance liquid chromatography-mass spectrometry as claimed in claim 1, wherein the edible oil sample is dissolved in n-hexane and diluted to 0.1-1.0 mg/mL by mobile phase.
4. The method for analyzing triglyceride composition of edible oil by HPLC-MS of claim 1, wherein the HPLC-MS is LCMS-2020 liquid mass spectrometer.
5. The method for analyzing triglyceride composition of edible oil by HPLC-MS as claimed in claim 1, wherein the edible oil sample comprises vegetable oil and animal oil.
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