CN109001358B - Method for determining small molecular compounds in olive oil - Google Patents
Method for determining small molecular compounds in olive oil Download PDFInfo
- Publication number
- CN109001358B CN109001358B CN201811074347.5A CN201811074347A CN109001358B CN 109001358 B CN109001358 B CN 109001358B CN 201811074347 A CN201811074347 A CN 201811074347A CN 109001358 B CN109001358 B CN 109001358B
- Authority
- CN
- China
- Prior art keywords
- olive oil
- time
- mass
- sample
- small molecular
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention provides a method for measuring small molecular compounds in olive oil, which adopts a real-time direct analysis and flight time mass spectrometry combined technology, can omit a pretreatment step, shortens the acquisition time of a sample, shortens the whole measurement process time by only 20-30 min, and can greatly improve the analysis efficiency; the mass spectrogram and the accurate mass number are combined and analyzed by software, so that the components of the small molecular compounds in the olive oil can be accurately determined, and the technology does not use any mobile phase in the analysis process, so that the solvent pollution can be reduced.
Description
Technical Field
The invention relates to the technical field of analysis and detection, in particular to a method for determining small molecular compounds in olive oil.
Background
The olive oil is prepared by directly cold pressing fresh olive fruits, and retains natural nutritional components without heating and chemical treatment. Olive oil is considered as the most suitable oil for human nutrition among the oils found so far. The quality of olive oil is the top in vegetable oil, so that the olive oil has the name of 'eating the queen of vegetable oil', and the olive oil can be seen to have an irreplaceable status in vegetable oil. In recent years, the quality problem of olive oil concerned by people has been raised from the past counterfeit and inferior to adulterated and the problem of origin of olive oil, so that the traditional method for identifying the authenticity of olive oil by measuring fatty acid has been increasingly challenged. And the identification of the small molecular compounds in the olive oil is the main research direction for identifying the authenticity and the production place of the olive oil.
Currently, small molecular compounds in olive oil mainly comprise organic phenolic substances, organic acids, bioflavonoids, terpenoids, other oxygen-containing compounds and the like, and the determination methods of the substances mainly comprise a potassium permanganate titration method, a Prussian blue method, a Folin-Denis colorimetric method, a Folin-Ciocalteus colorimetric method, a ferrous tartrate method, an ultraviolet spectrophotometer method, a gas chromatography-mass spectrometry combined method, a liquid chromatography-mass spectrometry combined method and the like. The sample pretreatment processes of the methods basically need complex processes such as sample weighing, extraction, degreasing, purification, concentration and the like, the time consumed for detecting a single sample is 2-3 hours, when a large number of samples are encountered, the detection of the item becomes the burden of a laboratory, the cost is high, the solvent consumption is large, and the detection of the large number of samples is difficult to complete in a short time. Moreover, without knowing the composition of the constituents of the olive oil under study, the identification of the compounds becomes a problem and will require a great deal of time and effort.
Disclosure of Invention
In view of the above, the present invention aims to provide a method for measuring small molecular compounds in olive oil. The determination method provided by the invention not only can accurately determine the components of the micromolecular compounds in the olive oil, but also can greatly shorten the operation time, and the time of the whole determination process is only 20-30 min.
In order to achieve the above object, the present invention provides the following technical solutions:
a method for measuring small molecular compounds in olive oil comprises the following steps:
(1) adding an olive oil sample into an organic solvent for extraction to obtain an extract;
(2) performing real-time direct analysis ionization treatment on the extract to obtain an ionized sample;
(3) performing time-of-flight mass spectrometry on the ionized sample to obtain a mass spectrogram, and obtaining the accurate mass number of a compound in the mass spectrogram through software;
(4) and (4) combining the mass spectrogram and the accurate mass number for analysis, and determining the name of the small molecular compound.
Preferably, the volume ratio of the olive oil sample to the organic solvent in the step (1) is 1: 10-5: 1; the organic solvent is acetonitrile and/or methanol.
Preferably, the extraction time in the step (1) is 1-30 min.
Preferably, the sample injection rate of the real-time direct analysis ionization treatment in the step (2) is 0.1-0.5 mm/s.
Preferably, the ionization temperature of the real-time direct analysis ionization treatment in the step (2) is 50-550 ℃, and the grid voltage is 50-530V.
Preferably, the ionized gas directly analyzed and ionized in the step (2) is helium, and the flow rate is 2-3L/min.
Preferably, the temperature of the ion source for the time-of-flight mass spectrometry in the step (3) is 80-150 ℃, the voltage of the taper hole is 10-60V, the collision energy of the collision cell 1 is 2-6V, the collision energy of the collision cell 2 is 2-6V, and the voltage of the detector is 1500-1900V.
Preferably, the mass spectrum and accurate mass number combined analysis in the step (4) specifically comprises: firstly, software is used for confirming the accurate mass number, then isotope matching is carried out, then the element composition is calculated out, and finally the name of the micromolecule compound is determined according to the element composition.
The invention provides a method for measuring small molecular compounds in olive oil, which adopts a real-time direct analysis and flight time mass spectrometry combined technology, can omit a pretreatment step, shortens the acquisition time of a sample, shortens the whole measurement process time by only 20-30 min, and can greatly improve the analysis efficiency; the mass spectrogram and the accurate mass number are combined and analyzed by software, so that the components of the small molecular compounds in the olive oil can be accurately determined, and the technology does not use any mobile phase in the analysis process, so that the solvent pollution can be reduced. The results of the examples show that the determination method provided by the invention can accurately detect organic phenolic substances, organic acids, bioflavonoids, terpenoids and other oxygen-containing compounds in olive oil, and the whole determination time is only 30 min.
Drawings
FIG. 1 is an enlarged mass spectrum of m/z 60-m/z 225 of an olive oil extract after real-time direct analysis ionization treatment and flight time mass spectrometry in example 1 of the present invention;
FIG. 2 is an enlarged mass spectrum of m/z 255-m/z 345 of the olive oil extract in example 1 of the present invention after real-time direct analysis ionization and time-of-flight mass spectrometry;
FIG. 3 is an enlarged mass spectrum of m/z345 to m/z487 of the olive oil extract in example 1 of the present invention after real-time direct analysis ionization and time-of-flight mass spectrometry;
FIG. 4 is a chromatogram of a liquid chromatography-electrospray-mass spectrometry combined assay of an olive oil extract of comparative example 1 of the present invention;
FIG. 5 is a mass spectrum of olive oil extract in comparative example 1 of the present invention in scan mode detected by liquid chromatography-electrospray-mass spectrometry.
Detailed Description
The invention provides a method for determining small molecular compounds in olive oil, which comprises the following steps:
(1) adding an olive oil sample into an organic solvent for extraction to obtain an extract;
(2) performing real-time direct analysis ionization treatment on the extract to obtain an ionized sample;
(3) performing time-of-flight mass spectrometry on the ionized sample to obtain a mass spectrogram, and obtaining the accurate mass number of a compound in the mass spectrogram through software;
(4) and (4) combining the mass spectrogram and the accurate mass number for analysis, and determining the name of the small molecular compound.
The invention adds an olive oil sample into an organic solvent for extraction to obtain an extract. The source of olive oil is not particularly required in the present invention, and commercially available or self-prepared olive oil can be measured using the method of the present invention. According to the method, the olive oil sample is preferably extracted in a vibration extraction mode, wherein the vibration extraction time is preferably 1-30 min, and more preferably 5-10 min; in the invention, the volume ratio of the olive oil sample to the organic solvent is preferably 1: 10-5: 1, and more preferably 1: 2-2: 1; in the present invention, the organic solvent is preferably acetonitrile and/or methanol.
According to the invention, the extract is preferably obtained by centrifuging and layering after shaking and collecting an organic solvent layer.
After obtaining the extract, the invention directly analyzes the extract in real time for ionization treatment (DART ionization treatment) to obtain an ionized sample. The DART ion source of IonSense company and SVP type is preferably used for DART ionization treatment; the invention preferably uses a liquid absorbing device to absorb the extract, then the extract point is put on a TIP head matched with a direct analysis ion source and then sample injection is carried out, and the TIP head with the sample is enabled to pass through a DART source sampling port in sequence. In the invention, the suction volume of the liquid suction device is preferably 1-10 muL, and more preferably 3-8 muL; the sampling rate is preferably 0.1-0.5 mm/s, and more preferably 0.2-0.4 mm/s; the sample injection mode is preferably a 12Dip-it Samplers mode. The invention uses the real-time direct analysis ionization treatment extract to replace the traditional pretreatment, and can greatly reduce the time of the whole process. In the invention, the ion source is directly analyzed in real time, and almost no adduct ions and extra-power ions are generated, so that each mass-to-charge ratio displayed on a spectrogram corresponds to a molecular ion peak of a certain compound, and the spectrogram is simple and convenient to analyze.
In the present invention, the mode of the real-time direct analysis ionization treatment is preferably a negative ion mode; the ionization temperature of the real-time direct analysis ionization treatment is preferably 50-550 ℃, more preferably 100-400 ℃, and most preferably 300 ℃; the grid voltage of the real-time direct analysis ionization treatment is preferably 50-530V, and more preferably 150-400V; the ionization gas for the real-time direct analysis ionization treatment is preferably helium, and the flow rate of the ionization gas is preferably 2-3L/min, and more preferably 2.2-2.8L/min.
After obtaining the ionized sample, the invention carries out flight time mass spectrometry on the ionized sample to obtain a mass spectrogram. The invention has no requirement on the brand and model of the time-of-flight mass spectrometer, and the time-of-flight mass spectrometer which is conventional in the field can be used. In the invention, the distance between the outlet of the real-time direct analysis ion source and the inlet of the time-of-flight mass spectrum is preferably 0.5-4 cm, and more preferably 2-3 cm; in the invention, the ion source temperature of the time-of-flight mass spectrometry is preferably 80-150 ℃, and more preferably 100-130 ℃; the taper hole voltage of the flight time mass spectrometry is preferably 10-60V, and more preferably 20-50V; the collision energy of the collision cell 1 for the time-of-flight mass spectrometry is preferably 2-6V, and more preferably 3-5V; the collision energy of the collision cell 2 for the time-of-flight mass spectrometry is preferably 2-6V, and more preferably 3-5V; the voltage of the detector for the time-of-flight mass spectrometry is preferably 1500-1900V, and more preferably 1600-1800V.
After obtaining the mass spectrogram, the invention obtains the accurate mass number of the compound in the mass spectrogram through software. In the present invention, the software preferably uses the Lockmass function in Masslynx software to obtain the exact mass number of the compound in the sample.
After the mass spectrogram and the accurate mass number are obtained, the mass spectrogram and the accurate mass number are combined for analysis, and the name of the small molecular compound is determined. In the present invention, the analysis is preferably specifically: firstly, software is used for confirming the accurate mass number, then isotope matching is carried out, then the element composition is calculated out, and finally the name of the micromolecule compound is determined according to the element composition. In the invention, the software used for confirming the accurate mass number is preferably Massworks software, the automatic calibration system of Massworks can obtain the accurate mass number of the compound, the accurate mass number is determined by crossing with a time-of-flight mass spectrometer, the accuracy and the reliability are higher, and meanwhile, the Massworks software can directly give the correct molecular formula of the compound parent ion or the plate ion by means of map accuracy, thereby reducing the matching range of the compound. The invention combines the spectrogram obtained by a mass spectrometer and accurate mass number with Massworks software, and can calculate the element composition after isotope matching.
After obtaining the elemental composition, the present invention preferably identifies the compound name on the Chemical spider website, matches the obtained compound name with the protocols reported in the relevant literature, and can finally confirm the composition.
The method for measuring small molecular compounds in olive oil according to the present invention will be described in detail with reference to the following examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Weighing 1g of Xiangyu olive oil sample, and shaking and extracting for 5 minutes by using 5mL of acetonitrile. Accurately sucking 2 mu L of extract points by using a liquid suction device, putting the extract points on a TIP head configured for implementing a direct analysis ion source, then injecting samples at a sample injection speed of 0.2mm/s, performing real-time direct analysis ionization treatment, entering a flight time mass spectrum for analysis to obtain a mass spectrogram, and obtaining the accurate mass number of the compound in the mass spectrogram through a Lockmass function in Masslynx software. And (3) calculating the element composition by combining the obtained spectrogram and the accurate mass number with Masswords software through isotope matching, finally determining the compound name on a Chemical spider website, matching with a scheme reported in Chinese literature in foreign olive oil, and finally determining the components. The measurement uses a real-time direct analysis ion source of IonSense company, model SVP, and a single quadrupole tandem time-of-flight mass spectrometer of Waters company, model Synrpt.
The parameters for this measurement of DART ionization treatment were: the ionized gas is helium, and the flow rate is 2 mL/min; the mode of DART ionization processing is negative ion mode; the sample injection mode is a 12Dip-it Samplers mode; the ionization temperature is 300 ℃; the grid voltage is 150V; the outlet of the ion source is 2.4cm away from the mass spectrum inlet.
The mass spectrum parameters of the measured flight time are as follows: the ion source temperature is 100 ℃; the voltage of the taper hole is 40V; the collision energy of the collision cell 1 is 6.0V; the collision energy of the collision cell 2 is 4.0V; the detector voltage was 1700V.
The amplified mass spectrum of m/z 60-m/z 225, the amplified mass spectrum of m/z 255-m/z 345 and the amplified mass spectrum of m/z 345-m/z 487, which are obtained after the olive oil extract is directly analyzed and ionized in real time and analyzed by flight time mass spectrometry, are shown in figure 1, figure 2 and figure 3 respectively. As can be seen from FIGS. 1 to 3, the mass spectrogram obtained by the method provided by the invention is simple and clear and is convenient to analyze.
The whole time of the measurement is 30min, and the structures of the measured substances and compounds are shown in the following table:
table 1 name and structure of small molecule compound determined in example 1
Comparative example 1
Weighing 1g of olive oil sample, shaking and extracting with 5mL of acetonitrile for 5 minutes, analyzing the extract by liquid chromatography-electrospray-mass spectrometry (HUPLC-ESI-Q-TOF), wherein the analysis time is 100 minutes, the obtained chromatogram is shown in figure 4, and the mass spectrum in the scan mode is shown in figure 5. As can be seen from fig. 4 to 5, the components in the olive oil are not completely separated, and due to the influence of external power ionization, the ESI source has a complex mass spectrum in the scan mode, cannot determine the molecular ion peak, is difficult to analyze, and cannot determine the final compound.
Comparing example 1 with comparative example 1, it can be seen that the method for determining the small molecular compounds in the olive oil provided by the invention not only can accurately determine the components of the small molecular compounds in the olive oil, but also can shorten the collection time of the sample and improve the analysis efficiency.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (4)
1. A method for measuring small molecular compounds in olive oil comprises the following steps:
(1) adding an olive oil sample into an organic solvent for extraction to obtain an extract; the organic solvent is acetonitrile;
(2) performing real-time direct analysis ionization treatment on the extract to obtain an ionized sample; the sample introduction rate of the real-time direct analysis ionization treatment is 0.1-0.5 mm/s;
the parameters of the real-time direct analysis ionization processing are as follows: the ionized gas is helium, and the flow rate is 2 mL/min; the mode of DART ionization processing is negative ion mode; the sample injection mode is a 12Dip-it Samplers mode; the ionization temperature is 300 ℃; the grid voltage is 150V; the distance between the outlet of the ion source and the mass spectrum inlet is 2.4 cm;
(3) performing time-of-flight mass spectrometry on the ionized sample to obtain a mass spectrogram, and obtaining the accurate mass number of a compound in the mass spectrogram through software;
the temperature of an ion source for flight time mass spectrometry is 80-150 ℃, the voltage of a taper hole is 10-60V, the collision energy of a collision pool 1 is 2-6V, the collision energy of a collision pool 2 is 2-6V, and the voltage of a detector is 1500-1900V;
(4) combining the mass spectrogram and the accurate mass number for analysis, and determining the name of the small molecular compound;
the small molecular compound comprises one or more of organic phenolic substances, organic acids, bioflavonoids and terpenoids.
2. The method according to claim 1, wherein the volume ratio of the olive oil sample to the organic solvent in the step (1) is 1:10 to 5: 1.
3. The method according to claim 1 or 2, wherein the extraction time in the step (1) is 1 to 30 min.
4. The assay method according to claim 1, wherein the mass spectrum and the precise mass number binding analysis in step (4) are specifically: firstly, software is used for confirming the accurate mass number, then isotope matching is carried out, then the element composition is calculated out, and finally the name of the micromolecule compound is determined according to the element composition.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811074347.5A CN109001358B (en) | 2018-09-14 | 2018-09-14 | Method for determining small molecular compounds in olive oil |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811074347.5A CN109001358B (en) | 2018-09-14 | 2018-09-14 | Method for determining small molecular compounds in olive oil |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109001358A CN109001358A (en) | 2018-12-14 |
| CN109001358B true CN109001358B (en) | 2021-03-09 |
Family
ID=64592329
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811074347.5A Active CN109001358B (en) | 2018-09-14 | 2018-09-14 | Method for determining small molecular compounds in olive oil |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109001358B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102393426A (en) * | 2011-10-28 | 2012-03-28 | 李涛 | Identification method for illegal cooking oil |
| WO2013166406A1 (en) * | 2012-05-04 | 2013-11-07 | Battelle Memorial Institute | Methods of distinguishing between similar compositions |
| CN104316592A (en) * | 2014-11-05 | 2015-01-28 | 西安石油大学 | Biological sample spectral analysis method based on solute migration electrospray ionization technique |
-
2018
- 2018-09-14 CN CN201811074347.5A patent/CN109001358B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102393426A (en) * | 2011-10-28 | 2012-03-28 | 李涛 | Identification method for illegal cooking oil |
| WO2013166406A1 (en) * | 2012-05-04 | 2013-11-07 | Battelle Memorial Institute | Methods of distinguishing between similar compositions |
| CN104316592A (en) * | 2014-11-05 | 2015-01-28 | 西安石油大学 | Biological sample spectral analysis method based on solute migration electrospray ionization technique |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109001358A (en) | 2018-12-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102565231B (en) | Method for determining eight kinds of alkaloid in cured tobacco through gas chromatography-nitrogen chemoluminescence detection method | |
| CN108072690B (en) | Ion mobility spectrometry and ion trap mass spectrometry combined device and analysis method | |
| CN102520092B (en) | Rapid detection method for discriminating adulteration of rice syrup in honey | |
| CN107144646B (en) | Analysis method for distinguishing true honey and syrup adulterated honey by applying liquid chromatography-mass spectrometry combined with metabonomics method | |
| CN102507800B (en) | Rapid aroma fingerprint identification method for geographical indication protection product of vinegar | |
| CN101601805B (en) | Method for detecting peimine and peiminine in Chinese medicament fritillaria extract | |
| CN110412183B (en) | Rose fragrance component analysis method by needle capture-gas chromatography-mass spectrometry | |
| Condrat et al. | Qualitative and quantitative analysis of gallic acid in Alchemilla vulgaris, Allium ursinum, Acorus calamus and Solidago virga-aurea by chip-electrospray ionization mass spectrometry and high performance liquid chromatography | |
| Chen et al. | Simultaneous determination of toxins in algae and water samples by high‐performance liquid chromatography with triple quadrupole mass spectrometry | |
| CN104237402B (en) | Extracting and measuring method of perfluorinated compounds in bark and leaves | |
| CN102354649A (en) | Surface extraction chemical ionization source and surface extraction chemical ionization mass spectrometry method | |
| CN107192770B (en) | Analytical method for identifying vitex negundo honey and syrup adulterated vitex negundo honey | |
| Du et al. | Simultaneous qualitative and quantitative analysis of 28 components in Isodon rubescens by HPLC‐ESI‐MS/MS | |
| CN108593756B (en) | Quick detection device of trace steroid in water | |
| CN109001358B (en) | Method for determining small molecular compounds in olive oil | |
| Xu et al. | Qualitative and quantitative determination of nine main active constituents in Pulsatilla cernua by high‐performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry | |
| CN107860858A (en) | A kind of method for high-flux analysis of mycotoxin in plant medicine material | |
| CN102507786A (en) | Structure identification method of components of traditional Chinese medicine | |
| Zhang et al. | Rapidly improved determination of metabolites from biological data sets using the high-efficient TransOmics tool | |
| Deng et al. | Quality assessment and origin tracing of Guangdong Liangcha granules using direct mass spectrometry fingerprinting | |
| RU2419788C2 (en) | Method of detecting unknown substances in body fluids of patients taking narcotic or psychoactive substances | |
| CN112697931A (en) | Application of trifolioside as characteristic marker of lespedeza-pedeza honey | |
| RU2390771C1 (en) | Method of identifying narcotic and psychoactive substances in biological fluids | |
| CN110609097A (en) | Method for screening phosphatidylserine compounds | |
| CN112285240B (en) | Detection method for three active ingredients of thunberg fritillary bulb in biological sample based on solid phase extraction-liquid chromatography-mass spectrometry |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information | ||
| CB02 | Change of applicant information |
Address after: 730000, No. 2168, Nanhe Road, Chengguan District, Lanzhou City, Gansu Province Applicant after: Lanzhou Customs Technical Center Address before: 730000, No. 2168, Nanhe Road, Chengguan District, Lanzhou City, Gansu Province Applicant before: INSPECTION AND QUARANTINE COMPREHENSIVE TECHNOLOGY CENTER, GANSU ENTRY-EXIT INSPECTION AND QUARANTINE BEREAU |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |