CN108152425B - Method for detecting lignanoids in sesame oil by high performance liquid chromatography - Google Patents
Method for detecting lignanoids in sesame oil by high performance liquid chromatography Download PDFInfo
- Publication number
- CN108152425B CN108152425B CN201810136159.4A CN201810136159A CN108152425B CN 108152425 B CN108152425 B CN 108152425B CN 201810136159 A CN201810136159 A CN 201810136159A CN 108152425 B CN108152425 B CN 108152425B
- Authority
- CN
- China
- Prior art keywords
- sesame oil
- high performance
- sesamin
- sesamolin
- performance liquid
- 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
-
- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/062—Preparation extracting sample from raw material
Abstract
A method for detecting lignans in sesame oil by high performance liquid chromatography is disclosed, wherein the lignans are sesamol, sesamin and sesamolin, and the method is characterized in that: the method comprises the steps of carrying out liquid-liquid microextraction on a phenolic eutectic solvent, carrying out centrifugal separation, directly detecting by using a high performance liquid chromatograph (HPLC-UV), and simultaneously determining the contents of sesamol, sesamin and sesamolin in sesame oil. Compared with the original sesame oil lignan analysis method, the analysis and determination method of the invention simplifies the sample pretreatment, has higher sensitivity, good experimental repeatability and recovery rate, and is suitable for the analysis of mass samples.
Description
Technical Field
The invention belongs to the technical field of food analysis and determination, and particularly relates to a method for detecting lignans in sesame oil by high performance liquid chromatography.
Background
The sesame oil is a special oil, and has good oxidation stability and long shelf life. The sesame oil contains sesamol, sesamin, sesamolin and other natural antioxidant components (the structural formula is shown in the specification), wherein the sesamol is a phenolic substance and has strong antioxidant property; while sesamin and sesamolin are not phenols per se, they are partially converted to sesamol during sesame oil processing and storage. In addition, the sesame oil from different sources has a large difference in the content of lignans. Therefore, in order to more comprehensively evaluate the quality of sesame oil and the variety of sesame oil, it is very important to simultaneously measure the contents of 3 kinds of lignans (sesamol, sesamin and sesamolin) in the sesame oil.
Structural formula of main lignans of sesame oil
Although a plurality of methods for analyzing and detecting the lignans of the sesames and the sesamols alone are reported, trace components in the sesame oil are very complicated and interfere a lot due to the raw material variety and different processing effects of the sesame oil, and the simultaneous analysis and determination of the lignans in the sesame oil still remains a challenging task. The current commonly used method is the national standard 'high performance liquid chromatography for measuring sesamin and sesamolin in sesame oil by grain and oil inspection' (GB/T31579-. The method comprises the steps of extracting, purifying and enriching a sample oil sample by using a silica gel solid phase extraction column, and quantitatively measuring the contents of sesamin and sesamolin in the obtained solution through a High Performance Liquid Chromatograph (HPLC) and an ultraviolet detector. The method is complex and time-consuming in sample pretreatment, and the pretreatment period of one sample is 4-6 hours. Wherein the solid phase extraction column needs to be activated firstly for use; in addition, the solid phase extraction and purification process consumes more time and uses a large amount of organic solvent, which seriously affects the analysis efficiency. Therefore, the method which is simple in sample pretreatment and higher in sensitivity is established, and has important significance for improving the analysis efficiency.
Disclosure of Invention
The invention aims to provide an analytical method based on eutectic solvent liquid-phase microextraction tandem high performance liquid chromatography, aiming at the problems of complex pretreatment, time consumption and large organic solvent consumption of a method for measuring lignans (sesamol, sesamin and sesamolin) in sesame oil in the prior art. The method comprises the steps of liquid-liquid extracting a grease sample by using a phenol eutectic solvent, centrifuging after extraction, directly detecting by using a high performance liquid chromatograph (HPLC-UV), and simultaneously determining the content of main lignans (sesamol, sesamin and sesamolin) in sesame oil.
The purpose of the invention is realized by the following scheme:
the method for determining lignans in sesame oil comprises the following steps:
a. sample extraction
Accurately weighing a certain amount of sesame oil in a triangular flask, dissolving with n-hexane, adding a certain amount of eutectic solvent, performing ultrasonic extraction, centrifuging at 3000 r/min for 10 min, and analyzing the eutectic solvent phase by a high performance liquid chromatograph.
b. Preparation of Standard solutions
Accurately weighing 0.10 g (accurate to 0.10 mg) of lignan in three different 100 mL brown volumetric flasks, dissolving with methanol (pure by chromatography) and diluting to 100 mL to obtain stock solution with lignan concentration of 1000 μ g/mL. Storing in a refrigerator at 4 deg.C. Transferring each lignan stock solution with different volumes into a 10 mL volumetric flask, diluting with methanol and fixing the volume to prepare the sesamol standard working solution, the sesamin standard working solution and the sesamolin standard working solution with the concentrations of 5, 10, 20, 50, 100, 200 and 500 mu g/mL. Mixing uniformly, respectively filtering with 0.45 μm organic filter membrane, sampling 20 μ L of sample, and establishing linear working curve of standard solution with peak area as ordinate and concentration as abscissa.
c. Analysis of the samples:
HPLC-UV analysis conditions:
ultraviolet detector, detector wavelength: 298 nm; 287 nm;
a chromatographic column: waters Symmetry C18 column (250 mm. times.5 μm. times.4.6 mm);
column temperature: 30 ℃; sample introduction amount: 20 μ L.
Mobile phase A: methanol, B: 0.5% acetic acid water, wherein a: 0 min-10 min, 30% -0%; 10-10.5 min, 0-30%; 10.5-19.5 min, 30%. 19.5-20 min, 30-80%; 20-30 min, 80%; 30-32 min, 80-30%; 32-35 min, 30%.
In the step a, the volume ratio of the sesame oil to the normal hexane to the eutectic solvent is 1: 1-10: 0.5-2.
When a sample is extracted, the ultrasonic temperature is 20-60 ℃, and the extraction time is 5-30 minutes.
The preparation process of the phenolic eutectic solvent is as follows: p-cresol (or m-cresol) and choline chloride (the molar ratio of the p-cresol to the m-cresol is 2: 1) are heated and mixed for 30 minutes in a round-bottom flask at the temperature of 80 ℃ to form uniform and transparent liquid, namely the phenolic eutectic solvent.
The following three standard samples of lignan compounds (sesamol, sesamin and sesamolin) were tested using the method of the invention, and the data such as compound retention times are shown in table 1:
liquid chromatography analysis parameters for the compounds of Table 1
| Compound (English name) | RT（min） |
| Sesamol (sesamol) | 17.8 |
| Sesamin (sesamin) | 27.0 |
| Sesame linn (sesamolin) | 28.0 |
The recovery rate test was carried out while the Relative Standard Deviation (RSD) of the results of 6 replicates in one day and 5 days with the same sesame oil was expressed as the day-to-day reproducibility and the day-to-day reproducibility of the method, and the results are shown in Table 2. Therefore, compared with the original analysis method, the method simplifies the pretreatment process of the sample, omits the purification step of the solid phase extraction column, has good repeatability and recovery rate of the experiment, and is suitable for the analysis of a large number of samples.
Table 2 repeatability and recovery test results of the method
| Compound (I) | sesamol | sesamin | sesamolin |
| RSD (in day%) | 0.74 | 0.67 | 0.48 |
| RSD (daytime,%) | 0.97 | 0.81 | 0.62 |
| Recovery (%) | 97.3 | 98.4 | 98.8 |
The invention has the following beneficial effects:
the key point of the method is that a phenolic eutectic solvent is used as a solvent for liquid phase microextraction, and the action mechanism of the method is that the phenolic eutectic solvent and various lignans in the sesame oil are interacted through pi-pi so as to realize simultaneous extraction. The invention adopts the liquid-liquid extraction of the phenol eutectic solvent to extract the grease sample, centrifugalizes the grease sample after extraction, directly adopts a high performance liquid chromatograph (HPLC-UV) to detect, and determines the content of main lignans (sesamol, sesamin and sesamolin) in the sesame oil, and the quantitative method is an external standard method, so that the pretreatment of the sample is simplified, and the accuracy and the repeatability are good. The method does not need to adopt a solid phase extraction column for purification, simultaneously omits the next concentration process, and effectively simplifies the sample pretreatment step.
Drawings
FIG. 1 is a chromatogram of a standard solution on an HPLC-UV instrument according to this invention.
Detailed Description
The invention will be further illustrated with reference to the following examples:
example 1
Adding 400 mu L of phenolic eutectic solvent (the molar ratio of p-cresol to choline chloride is 2: 1) into a triangular flask containing 0.2 g of sesame oil, adding 3 mL of n-hexane, and extracting at 50 ℃ for 30 minutes. Centrifuging at 3000 r/min for 10 min, and analyzing with HPLC (high performance liquid chromatography) by taking phenol eutectic solvent (100 μ L).
HPLC-UV chromatography conditions, UV detector, detector wavelength: 298 nm; 287 nm.
A chromatographic column: waters Symmetry C18 column (250 mm. times.5 μm. times.4.6 mm); column temperature: 30 ℃; sample introduction amount: 20 μ L. Mobile phase A: methanol, B: 0.5% acetic acid water, wherein a: 0 min-10 min, 30% -0%; 10-10.5 min, 0-30%; 10.5-19.5 min, 30%. 19.5-20 min, 30-80%; 20-30 min, 80%; 30-32 min, 80-30%; 32-35 min, 30%.
The results of the measurements on 7 brands of sesame oil samples are shown in the following table:
| sesame oil sample | Sesamol (mg/kg) | Sesamin (mg/kg) | Sesamolin (mg/kg) |
| 1 | 51.62 | 2704.06 | 1175.89 |
| 2 | 54.30 | 2396.28 | 709.31 |
| 3 | 85.00 | 2439.66 | 937.79 |
| 4 | 130.42 | 2681.43 | 855.51 |
| 5 | 68.27 | 2076.85 | 665.65 |
| 6 | 62.48 | 2042.62 | 778.36 |
| 7 | 59.13 | 2075.67 | 617.47 |
Example 2
Adding 100 mu L of phenolic eutectic solvent (the molar ratio of m-cresol to choline chloride is 2: 1) into a triangular flask containing 0.2 g of blend oil (containing sesame oil), adding 1mL of n-hexane, and extracting at 30 ℃ for 20 minutes. Centrifuging at 3000 r/min for 10 min, and analyzing with HPLC (high performance liquid chromatography) by taking phenol eutectic solvent (100 μ L).
HPLC-UV chromatography conditions, UV detector, detector wavelength: 298 nm; 287 nm.
A chromatographic column: waters Symmetry C18 column (250 mm. times.5 μm. times.4.6 mm); column temperature: 30 ℃; sample introduction amount: 20 μ L. Mobile phase A: methanol, B: 0.5% acetic acid water, wherein a: 0 min-10 min, 30% -0%; 10-10.5 min, 0-30%; 10.5-19.5 min, 30%. 19.5-20 min, 30-80%; 20-30 min, 80%; 30-32 min, 80-30%; 32-35 min, 30%.
The results of measurements on 3 brands of blend oil (containing sesame oil) samples are shown in the following table:
| blend oil samples | Sesamol (mg/kg) | Sesamin (mg/kg) | Sesamolin (mg/kg) |
| 1 | 2.95 | 64.75 | 24.82 |
| 2 | 4.78 | 127.35 | 46.70 |
| 3 | 5.77 | 165.11 | 60.49 |
Claims (5)
1. A method for detecting lignans in sesame oil by high performance liquid chromatography is disclosed, wherein the lignans are sesamol, sesamin and sesamolin, and the method is characterized in that: after liquid-liquid microextraction of a phenolic eutectic solvent, performing centrifugal separation, directly detecting by adopting HPLC-UV, and simultaneously determining the contents of sesamol, sesamin and sesamolin in sesame oil, the method comprises the following specific steps:
a. sample extraction
Accurately weighing a certain amount of sesame oil in a triangular flask, dissolving with n-hexane, adding a certain amount of phenolic eutectic solvent, performing ultrasonic extraction, centrifuging at 3000 r/min for 10 min, and subjecting the eutectic solvent to high performance liquid chromatography; the phenolic eutectic solvent is prepared from p-cresol or m-cresol and choline chloride, and the molar ratio of the p-cresol or m-cresol to the choline chloride is as follows: 2: 1;
b. preparation of a standard solution: respectively preparing standard working solutions of sesamol, sesamin and sesamolin by using methanol, wherein the concentration gradient is 5, 10, 20, 50, 100, 200 and 500 mu g/mL; respectively filtering with 0.45 μm organic filter membrane, sampling 20 μ L of sample, and establishing linear working curve of standard solution with peak area as ordinate and concentration as abscissa;
c. analysis of the samples:
HPLC-UV analysis conditions:
ultraviolet detector, detector wavelength: 298 nm; 287 nm;
a chromatographic column: waters Symmetry C18 column, specification 250 mm × 5 μm × 4.6 mm;
column temperature: 30 ℃; sample introduction amount: 20 mu L of the solution;
mobile phase A: methanol, B: 0.5% acetic acid water, wherein a: 0 min-10 min, 30% -0%; 10-10.5 min, 0-30%; 30% in 10.5-19.5 min; 19.5-20 min, 30-80%; 20-30 min, 80%; 30-32 min, 80-30%; 32-35 min, 30%.
2. The method for detecting lignans in sesame oil by high performance liquid chromatography according to claim 1, wherein the method comprises the following steps: in the step a, the volume ratio of the sesame oil to the normal hexane to the eutectic solvent is 1: 1-10: 0.5-2.
3. The method for detecting lignans in sesame oil by high performance liquid chromatography according to claim 1, wherein the method comprises the following steps: in the step a, the ultrasonic extraction temperature is 20-60 ℃, and the extraction time is 5-30 minutes.
4. The method for detecting lignans in sesame oil by high performance liquid chromatography according to claim 1, wherein the method comprises the following steps: the preparation process of the phenolic eutectic solvent is as follows: heating and mixing p-cresol or m-cresol and choline chloride in a round-bottom flask at 80 ℃ for 30 minutes to form uniform and transparent liquid, namely the phenol eutectic solvent.
5. The method for detecting lignans in sesame oil by high performance liquid chromatography according to claim 1, wherein the method comprises the following steps: the specific preparation method of the standard working solution is as follows: accurately weighing sesamol, sesamin and sesamolin respectively 0.10 g in three different 100 mL brown volumetric flasks, dissolving with methanol respectively, and metering to 100 mL to obtain sesame powder stock solution, sesamin stock solution and sesamolin stock solution with concentration of 1000 μ g/mL; storing in a refrigerator at 4 deg.C; transferring each stock solution with different volumes into a 10 mL volumetric flask, diluting with methanol and fixing the volume to prepare sesamol standard working solution, sesamin standard working solution and sesamolin standard working solution with the concentrations of 5, 10, 20, 50, 100, 200 and 500 mu g/mL.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810136159.4A CN108152425B (en) | 2018-02-09 | 2018-02-09 | Method for detecting lignanoids in sesame oil by high performance liquid chromatography |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810136159.4A CN108152425B (en) | 2018-02-09 | 2018-02-09 | Method for detecting lignanoids in sesame oil by high performance liquid chromatography |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108152425A CN108152425A (en) | 2018-06-12 |
| CN108152425B true CN108152425B (en) | 2020-10-09 |
Family
ID=62457258
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810136159.4A Active CN108152425B (en) | 2018-02-09 | 2018-02-09 | Method for detecting lignanoids in sesame oil by high performance liquid chromatography |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108152425B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109001332A (en) * | 2018-09-06 | 2018-12-14 | 河南工业大学 | The measuring method of the original additive amount of TBHQ in a kind of edible oil and fat |
| CN112255339B (en) * | 2020-10-12 | 2021-05-25 | 南京海关动植物与食品检测中心 | Method for rapidly determining contents of three sesame lignans in sesame oil |
| CN115060840A (en) * | 2022-07-26 | 2022-09-16 | 吉首大学 | HPLC quality control method for white flower paulownia leaf sesame oil preparation |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2006119017A (en) * | 2004-10-22 | 2006-05-11 | Oki Electric Ind Co Ltd | Method for determining type of plant using thermal cracking gas chromatography |
| CN103389342B (en) * | 2012-05-08 | 2014-09-03 | 上海良友（集团）有限公司 | Method for detecting where leached sesame oil is doped in pressed sesame oil |
| CN104792890B (en) * | 2015-03-25 | 2017-01-04 | 河南省农业科学院芝麻研究中心 | The method of sesamin content in mensuration sesame seed |
| CN104777246A (en) * | 2015-03-25 | 2015-07-15 | 河南省农业科学院芝麻研究中心 | Method for determining content of sesamin in sesame |
| KR101823020B1 (en) * | 2016-03-24 | 2018-01-29 | 한국식품연구원 | Analytical method to detect α-dicarbonyl compounds, including glyoxal, methylglyoxal and diacetyl |
-
2018
- 2018-02-09 CN CN201810136159.4A patent/CN108152425B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN108152425A (en) | 2018-06-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108152425B (en) | Method for detecting lignanoids in sesame oil by high performance liquid chromatography | |
| CN109884207B (en) | Method for rapidly and accurately analyzing polyphenol content in rapeseed oil | |
| CN106290682B (en) | Chiral analysis method of nicotine in tea | |
| CN109696499B (en) | High-sensitivity determination method for nitrosamine in water based on high-resolution mass spectrum | |
| CN107655991B (en) | Method for measuring 6 kinds of phthalate in soil | |
| CN110632203A (en) | Synchronous and rapid detection of vitamin A and vitamin D3And vitamin E | |
| CN109030658B (en) | Method for detecting fructo-oligosaccharide and raffinose in infant milk powder | |
| Liu et al. | Elimination of matrix effects for headspace solid-phase microextraction of important volatile compounds in red wine using a novel coating | |
| CN111458442B (en) | Method for measuring content of conjugated estrogen in pregnant mare urine | |
| CN107389811B (en) | Method for measuring diisopropyl naphthalene in cigarette tipping paper | |
| CN112526007A (en) | Method for separating and detecting contents of m-cresol and p-cresol by using ultra-high liquid chromatography and application | |
| CN113533548B (en) | Method for detecting 1-vinyl imidazole in chemical product | |
| CN106483230B (en) | A kind of rapid detection method of urine Hydroxyl Polycyclic Aromatic | |
| CN111965273A (en) | HPLC (high Performance liquid chromatography) method for detecting genotoxic impurities in candesartan cilexetil | |
| CN110895264A (en) | Method for determining ethyl bromide in tenofovir alafenamide | |
| CN109633015B (en) | Method for detecting additive in entolipide | |
| Xing et al. | Graphene‐sensitized microporous membrane/solvent microextraction for the preconcentration of cinnamic acid derivatives in Rhizoma Typhonii | |
| CN114609295A (en) | High performance liquid chromatography analysis method for quinic acid content in tala enzymolysis waste liquid | |
| Xu et al. | Capillary liquid chromatographic analysis of fat-soluble vitamins and β-carotene in combination with in-tube solid-phase microextraction | |
| CN108398506B (en) | Method for splitting and measuring enantiomers of chiral pesticides quizalofop-ethyl and fluazifop-p-butyl by ultra-high performance combined chromatography-tandem mass spectrometry technology | |
| CN112782318A (en) | Method for detecting doxorubicin hydrochloride by using HPLC (high performance liquid chromatography) | |
| CN111579675A (en) | Method for detecting fat-soluble vitamins in feed | |
| CN112213417A (en) | Kit and method for detecting concentration of mycophenolic acid medicine in dried blood spots | |
| CN112255339A (en) | Method for rapidly determining contents of three sesame lignans in sesame oil | |
| CN111024868B (en) | Method for detecting content of alpha-high nojirimycin in white tree medicinal material |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |