CN112697925A - High performance liquid chromatography detection method for matrine pesticide residue in fruits and vegetables - Google Patents
High performance liquid chromatography detection method for matrine pesticide residue in fruits and vegetables Download PDFInfo
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- ZSBXGIUJOOQZMP-JLNYLFASSA-N Matrine Chemical compound C1CC[C@H]2CN3C(=O)CCC[C@@H]3[C@@H]3[C@H]2N1CCC3 ZSBXGIUJOOQZMP-JLNYLFASSA-N 0.000 title claims abstract description 85
- 229930014456 matrine Natural products 0.000 title claims abstract description 85
- 238000001514 detection method Methods 0.000 title claims abstract description 52
- 238000004128 high performance liquid chromatography Methods 0.000 title claims abstract description 45
- 235000012055 fruits and vegetables Nutrition 0.000 title claims abstract description 23
- 239000000447 pesticide residue Substances 0.000 title claims abstract description 19
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- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 57
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- 239000012488 sample solution Substances 0.000 claims abstract description 16
- 239000012086 standard solution Substances 0.000 claims abstract description 16
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- 239000013582 standard series solution Substances 0.000 claims abstract description 9
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 30
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- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 7
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Abstract
The invention relates to a high performance liquid chromatography detection method for matrine pesticide residue in fruits and vegetables, which comprises the following steps: (1) extracting matrine in a sample to be detected by using acetonitrile aqueous solution, purifying, extracting and redissolving the obtained extracting solution by using a strong cation exchange extraction column to obtain sample solution; (2) preparing a standard solution of matrine, and preparing a plurality of standard series solutions according to concentration gradient; (3) detecting and analyzing the sample solution and the standard solution by high performance liquid chromatography, and obtaining the content of matrine by an external standard method. The invention has the beneficial effects that: can be more targeted, and can quickly and effectively carry out accurate analysis and detection on matrine pesticide residue in fruits and vegetables, so that the component detection and analysis are quicker. The obtained matrine has good peak shape and separation degree, good linearity, and can be effectively separated from other impurities in the matrix, and the detection and extraction are more accurate. The process is simple, the extraction efficiency is high, the purification effect is good, the detection precision is high, and the detection is sensitive.
Description
Technical Field
The invention belongs to the technical field of food pollutant detection, and particularly relates to a high performance liquid chromatography detection method for matrine pesticide residues in fruits and vegetables.
Background
The matrine is prepared by extracting dried roots, plants and fruits of sophora flavescens ait of leguminous plants by using organic solvents such as ethanol, is an alkaloid, is a low-toxicity and environment-friendly pesticide in agriculture, has multiple functions of killing insects, sterilizing activity, regulating plant growth and the like, and has better effects on preventing and controlling plant diseases and insect pests and regulating plant growth, so that the matrine serving as an active ingredient of a novel plant-derived pesticide is widely applied to the planting and cultivating process of crops. The pesticide can not only be attached to crops, but also enter soil, atmosphere and water, and irrigation and precipitation bring residual drugs in the soil and the atmosphere into the water, so that passive plant enrichment is easy to realize, and human health can be endangered through the transmission of a food chain. The matrine has good prevention and treatment effect on fruit and vegetable diseases, but the detection method standard of the matrine in fruits and vegetables is not existed in GB2763-2019 'maximum pesticide residue limit in national food standard for food safety', most of the literatures aim at the detection of traditional Chinese medicines and tobacco, and fruit and vegetable products are not involved. However, the phenomena of the use of the matrine beyond the range and the addition of the matrine beyond the limit exist. Developed countries have implemented technical barriers to the trade of vegetables, fruits and aquatic products in China by pesticide limits. At present, the detection of matrine suitable for use internationally is mainly the residual quantity of matrine by liquid chromatography tandem mass spectrometry and gas chromatography tandem mass spectrometry, and the detection method has the disadvantages of more complicated pretreatment steps, longer detection time, expensive equipment and larger consumed reagent amount. In addition, the fruit and vegetable products contain more pigments and organic acids, the confirmation of a target peak is interfered to different degrees, and a solid phase extraction mode is adopted to extract and purify a sample, so that the interference is reduced.
Therefore, the method for rapidly determining the content of the matrine in the fruits and vegetables by establishing the solid-phase extraction-high performance liquid chromatography can be beneficial to the operation of large-scale detection of the matrine in the fruits and vegetables, greatly improves the analysis efficiency of samples, reduces the detection difficulty, improves the accuracy of sample detection, and better monitors the quality safety and the effectiveness of the fruits and vegetables.
Disclosure of Invention
In order to solve the problems, the invention provides a high performance liquid chromatography detection method for matrine pesticide residue in fruits and vegetables, which not only reduces cost, but also reduces interference and ensures detection accuracy, and the specific method comprises the following steps:
a high performance liquid chromatography detection method for matrine pesticide residue in fruits and vegetables comprises the following steps: extracting matrine in a sample to be detected by adopting a solution with the volume ratio of acetonitrile to water being 9:1, purifying, extracting and redissolving the obtained extracting solution by using a strong cation exchange extraction column to obtain a sample solution; the method comprises the following specific steps: accurately weighing 20g of a sample to be detected to be accurate to 0.001g, placing the sample into a centrifuge tube, accurately adding 50mL of a solution with a volume ratio of acetonitrile to water of 9:1, setting the frequency of 250 times/min and vigorous shaking for 10-15min, placing the sample into a centrifuge after standing, centrifuging the sample at 3000r/min for 5min, taking supernatant, continuously extracting residues with a solution with a volume ratio of acetonitrile to water of 25mL of 9:1, combining filtrate in a 100mL volumetric flask, adding 4mL of hydrochloric acid of 1.0mol/L, fixing the volume with the solution with the volume ratio of acetonitrile to water of 9:1, and taking out 5mL of the filtrate for purification; activating the strong cation exchange extraction column by using 5% ammoniated methanol solution, pre-washing by using methanol and water in sequence, after 5mL of filtrate passes through the column, washing the strong cation exchange extraction column by using water and methanol in sequence, and draining; eluting the extraction column twice with 5% ammoniated methanol solution, drying the eluent at 40 deg.C with nitrogen, adding 1.00mL of mobile phase, vortex for 30s, and passing through 0.22 μm organic filter membrane for instrument measurement;
preparing a standard solution of matrine, and preparing a plurality of standard series solutions according to concentration gradient;
detecting and analyzing the sample solution and the standard solution by high performance liquid chromatography, and obtaining the content of matrine by an external standard method.
Preferably, the preparation of the standard series of solutions comprises the following steps: precisely weighing appropriate amount of matrine standard substance, dissolving and diluting with methanol, transferring to volumetric flask and fixing volume to obtain matrine standard stock solution with appropriate concentration, respectively sucking appropriate amount of stock solution, and making into serial standard working solutions with concentration of 0.1ug/ml, 0.3ug/ml, 0.5ug/ml, 1.0ug/ml, 5.0ug/ml and 10.0ug/ml by flow matching.
Preferably, the detection analysis by high performance liquid chromatography comprises the following steps: respectively injecting the standard working solution and the sample working solution into a high performance liquid chromatograph, performing high performance liquid chromatography analysis, quantitatively comparing the peak area of the standard working solution with the peak area of the sample working solution, retaining time for qualitative determination, calculating by an external standard method, and calculating the content of matrine in the sample; the chromatographic conditions of the high performance liquid chromatography are as follows:
a chromatographic column: c18A chromatographic column with the specification of 150mm of column length, 4.6mm of inner diameter and 5 mu m of particle size; column temperature: 30 +/-5 ℃;
sample introduction amount: 10 mu L of the solution; the mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is methanol, the mobile phase B is a 5% disodium hydrogen phosphate aqueous solution with the pH value of 9, and the volume ratio of the mobile phase A to the mobile phase B is 40: 60; flow rate: 1.0 mL/min; detection wavelength: the wavelength was set at 215 nm.
Matrine belongs to water-soluble pesticide, and has high solubility in alkaline solution. The test compares different kinds and mixing ratios of the extracts of methanol, water-methanol, acetonitrile, ammonia-acetonitrile, acetonitrile-water, etc. The recovery rate is not high by using methanol and water-methanol extraction; extracting organic phase with acetonitrile to reduce impurities, but residual matrine in water phase to reduce recovery rate; the extraction solutions of ammonia water and acetonitrile in different proportions are used, and the recovery rate of the ammonia water and the acetonitrile to individual vegetables is not stable; the extracting solutions with different acetonitrile-water ratios are used, when the volume ratio of acetonitrile-water is 9:1, the acetonitrile has strong polarity, can effectively precipitate components such as protein and the like, reduces interference, has high extraction rate of organic phase matrine, ensures high solubility of matrine, achieves better extraction effect, and has high and stable recovery rate. Therefore, the matrine is ensured to be completely extracted by using the acetonitrile-water 9:1 mixed solution for two times.
Because the fruit and vegetable samples contain more pigments, fatty acids, saccharides, fruit acids and other substances and can interfere with a target peak, the Bond Elut Plexa PCX strong cation exchange extraction column selected in the patent has a good extraction effect on matrine, can adsorb the organic acids, fruit acids, pigments and other substances, and has less interference impurities in the extracting solution, a good extraction effect and a high sample recovery rate. Adding 4ml of 1.0mol/L hydrochloric acid into the sample extracting solution, and controlling the pH value of the extracting solution to be 2-4 so as to ensure that the extracting solution has enough charges and is completely retained on a small column before elution; before sampling, the strong cation exchange extraction column is fully activated by 5% ammoniated methanol solution, so that the matrine can be effectively adsorbed on the small column, and the method can effectively improve the recovery rate to more than 80%.
The high performance liquid chromatography adopts isocratic elution. The mobile phase with the proportion has better peak shape and shortest balancing time, can eliminate the interference from other components, and can better separate the target component from other components in the sample.
Through methodology examination, the matrine sample amount is in the range of 0.10-10.0 mug/mL, and has a good linear relation with the peak area (Y (area) ═ 107.2x (concentration) — 1.02), and all correlation coefficients are larger than 0.9996. The detection limit was 0.1 mg/kg.
The invention has the beneficial effects that:
1. the high performance liquid chromatography detection method for matrine pesticide residues in fruits and vegetables provided by the invention can be used for more pertinently, quickly and effectively carrying out accurate analysis and detection on matrine pesticide residues in fruits and vegetables, so that the component detection and analysis are quicker, and the time is saved.
2. The high performance liquid chromatography detection method for matrine pesticide residues in fruits and vegetables provided by the invention has the advantages that the obtained matrine has good peak shape and separation degree, is good in linearity, can be effectively separated from other impurities in a matrix, avoids interference of other substances, and is more accurate in detection and extraction.
3. The pretreatment method disclosed by the invention is simple in process, high in extraction efficiency, wide in application range of SPE (solid phase extraction) small columns used for pretreatment, good in purification effect and friendly to people and environment by adopting reagents. The recovery rate, precision and detection limit of the method are within the range required by the method, the experimental requirements can be met even in the actual sample measurement, the relative deviation is within the range required by the standard, and the daily inspection can be met.
Drawings
FIG. 1 liquid chromatogram for detecting matrine in potato with addition level of 1.0mg/kg
Detailed description of the invention
The invention will be better illustrated by the following examples without limiting the invention thereto.
Our reagent and apparatus
Reagent methanol: chromatography; disodium hydrogen phosphate: an analysis stage; hydrochloric acid; ammonia water; 5% ammoniated methanol solution: 5 portions of ammonia water are slowly added into 100mL of methanol solution and mixed evenly.
The instrument comprises the following steps: agilent 1260 Agilent high performance liquid chromatograph (ultraviolet detector)
Example 1
A high performance liquid chromatography detection method for matrine pesticide residue in cucumber comprises the following steps:
(1) extracting matrine in a sample to be detected by adopting a solution with the volume ratio of acetonitrile to water being 9:1, purifying, extracting and redissolving the obtained extracting solution by using a strong cation exchange extraction column to obtain a sample solution; the method comprises the following specific steps: accurately weighing 20g of a sample to be detected, placing the sample into a centrifuge tube, accurately adding 50mL of acetonitrile-water volume ratio 9:1 solution, setting the frequency of 250 times/min to shake for 15min violently, placing the sample into a centrifuge after standing, centrifuging for 5min at 3000r/min, taking supernatant, continuously extracting residues with 25mL of acetonitrile-water volume ratio 9:1 solution, combining filtrates in a 100mL volumetric flask, adding 4mL of 1.0mol/L hydrochloric acid, fixing the volume with acetonitrile-water volume ratio 9:1 solution, and taking out 5mL of filtrate for purification; activating the strong cation exchange extraction column by using 5% ammoniated methanol solution, pre-washing by using methanol and water in sequence, after 5mL of filtrate passes through the column, washing the strong cation exchange extraction column by using water and methanol in sequence, and draining; eluting the extraction column twice with 5% ammoniated methanol solution, drying the eluent at 40 deg.C with nitrogen, adding 1.00mL of mobile phase, vortex for 30s, and passing through 0.22 μm organic filter membrane for instrument measurement;
(2) preparing a standard solution of matrine, and preparing a plurality of standard series solutions according to concentration gradient; precisely weighing appropriate amount of matrine standard substance, dissolving and diluting with methanol, transferring to volumetric flask and fixing volume to obtain matrine standard stock solution with appropriate concentration, respectively sucking appropriate amount of stock solution, and making into serial standard working solutions with concentration of 0.1ug/ml, 0.3ug/ml, 0.5ug/ml, 1.0ug/ml, 5.0ug/ml and 10.0ug/ml by flow matching.
(3) Detecting and analyzing the sample solution and the standard solution by adopting a high performance liquid chromatography, obtaining the content of the matrine by utilizing an external standard method, respectively injecting a standard working solution and a sample working solution into a high performance liquid chromatograph, carrying out high performance liquid chromatography analysis, carrying out quantitative comparison on the peak area of the standard working solution and the peak area of the sample working solution, retaining time for qualitative determination, calculating by adopting the external standard method, and calculating the content of the matrine in the sample; the method is characterized in that the chromatographic conditions of the high performance liquid chromatography are as follows:
a chromatographic column: c18 chromatographic column with column length of 150mm, inner diameter of 4.6mm and particle size of 5 μm; column temperature: 30 +/-5 ℃;
sample introduction amount: 10 mu L of the solution; the mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is methanol, the mobile phase B is a 5% disodium hydrogen phosphate aqueous solution with the pH value of 9, and the volume ratio of the mobile phase A to the mobile phase B is 40: 60; flow rate: 1.0 mL/min; detection wavelength: the wavelength was set at 215 nm.
Example 2
A high performance liquid chromatography detection method for matrine pesticide residue in potatoes is characterized by comprising the following steps:
(1) extracting matrine in a sample to be detected by adopting a solution with the volume ratio of acetonitrile to water being 9:1, purifying, extracting and redissolving the obtained extracting solution by using a strong cation exchange extraction column to obtain a sample solution; the method comprises the following specific steps: accurately weighing 20g of a sample to be detected, placing the sample into a centrifuge tube, accurately adding 50mL of acetonitrile-water volume ratio 9:1 solution, setting the frequency of 300 times/min to shake vigorously for 12min, placing the sample into a centrifuge after standing, centrifuging the sample at 3000r/min for 5min, taking supernatant, continuously extracting residues with 25mL of acetonitrile-water volume ratio 9:1 solution, combining filtrates in a 100mL volumetric flask, adding 4mL of 1.0mol/L hydrochloric acid, performing constant volume with acetonitrile-water volume ratio 9:1 solution, and taking out 5mL of filtrate for purification; activating the strong cation exchange extraction column by using 5% ammoniated methanol solution, pre-washing by using methanol and water in sequence, after 5mL of filtrate passes through the column, washing the strong cation exchange extraction column by using water and methanol in sequence, and draining; eluting the extraction column twice with 5% ammoniated methanol solution, drying the eluent at 40 deg.C with nitrogen, adding 1.00mL of mobile phase, vortex for 30s, and passing through 0.22 μm organic filter membrane for instrument measurement;
(2) preparing a standard solution of matrine, and preparing a plurality of standard series solutions according to concentration gradient; precisely weighing appropriate amount of matrine standard substance, dissolving and diluting with methanol, transferring to volumetric flask and fixing volume to obtain matrine standard stock solution with appropriate concentration, respectively sucking appropriate amount of stock solution, and making into serial standard working solutions with concentration of 0.1ug/ml, 0.3ug/ml, 0.5ug/ml, 1.0ug/ml, 5.0ug/ml and 10.0ug/ml by flow matching.
(3) Detecting and analyzing the sample solution and the standard solution by adopting a high performance liquid chromatography, and obtaining the content of the matrine by utilizing an external standard method, wherein the high performance liquid chromatography detection and analysis comprises the following steps: respectively injecting the standard working solution and the sample working solution into a high performance liquid chromatograph, performing high performance liquid chromatography analysis, quantitatively comparing the peak area of the standard working solution with the peak area of the sample working solution, retaining time for qualitative determination, calculating by an external standard method, and calculating the content of matrine in the sample; the method is characterized in that the chromatographic conditions of the high performance liquid chromatography are as follows:
a chromatographic column: c18 chromatographic column with column length of 150mm, inner diameter of 4.6mm and particle size of 5 μm; column temperature: 30 +/-5 ℃;
sample introduction amount: 10 mu L of the solution; the mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is methanol, the mobile phase B is a 5% disodium hydrogen phosphate aqueous solution with the pH value of 9, and the volume ratio of the mobile phase A to the mobile phase B is 40: 60; flow rate: 1.0 mL/min; detection wavelength: the wavelength was set at 215 nm.
Example 3
A high performance liquid chromatography detection method for matrine pesticide residue in Chinese cabbage is characterized by comprising the following steps:
(1) extracting matrine in a sample to be detected by adopting a solution with the volume ratio of acetonitrile to water being 9:1, purifying, extracting and redissolving the obtained extracting solution by using a strong cation exchange extraction column to obtain a sample solution; the method comprises the following specific steps: accurately weighing 20g of a sample to be detected, placing the sample into a centrifuge tube, accurately adding 50mL of acetonitrile-water volume ratio 9:1 solution, setting the frequency of 400 times/min to shake for 10min violently, placing the sample into a centrifuge after standing, centrifuging for 5min at 3000r/min, taking supernatant, continuously extracting residues with 25mL of acetonitrile-water volume ratio 9:1 solution, combining filtrates in a 100mL volumetric flask, adding 4mL of 1.0mol/L hydrochloric acid, fixing the volume with acetonitrile-water volume ratio 9:1 solution, and taking out 5mL of filtrate for purification; activating the strong cation exchange extraction column by using 5% ammoniated methanol solution, pre-washing by using methanol and water in sequence, after 5mL of filtrate passes through the column, washing the strong cation exchange extraction column by using water and methanol in sequence, and draining; eluting the extraction column twice with 5% ammoniated methanol solution, drying the eluent at 40 deg.C with nitrogen, adding 1.00mL mobile phase, vortex for 30s, and filtering with 0.22 μm organic filter membrane for instrument determination
2) Preparing a standard solution of matrine, and preparing a plurality of standard series solutions according to concentration gradient; precisely weighing appropriate amount of matrine standard substance, dissolving and diluting with methanol, transferring to volumetric flask and fixing volume to obtain matrine standard stock solution with appropriate concentration, respectively sucking appropriate amount of stock solution, and making into serial standard working solutions with concentration of 0.1ug/ml, 0.3ug/ml, 0.5ug/ml, 1.0ug/ml, 5.0ug/ml and 10.0ug/ml by flow matching.
(3) Detecting and analyzing the sample solution and the standard solution by adopting a high performance liquid chromatography, and obtaining the content of the matrine by utilizing an external standard method, wherein the high performance liquid chromatography detection and analysis comprises the following steps: respectively injecting the standard working solution and the sample working solution into a high performance liquid chromatograph, performing high performance liquid chromatography analysis, quantitatively comparing the peak area of the standard working solution with the peak area of the sample working solution, retaining time for qualitative determination, calculating by an external standard method, and calculating the content of matrine in the sample; the method is characterized in that the chromatographic conditions of the high performance liquid chromatography are as follows:
a chromatographic column: c18 chromatographic column with column length of 150mm, inner diameter of 4.6mm and particle size of 5 μm; column temperature: 30 +/-5 ℃;
sample introduction amount: 10 mu L of the solution; the mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is methanol, the mobile phase B is a 5% disodium hydrogen phosphate aqueous solution with the pH value of 9, and the volume ratio of the mobile phase A to the mobile phase B is 40: 60; flow rate: 1.0 mL/min; detection wavelength: the wavelength was set at 215 nm.
Example 4
A high performance liquid chromatography detection method for matrine pesticide residue in apples comprises the following steps:
(1) extracting matrine in the apples to be detected by adopting a solution with the volume ratio of acetonitrile to water being 9:1, purifying, extracting and redissolving the obtained extracting solution by using a strong cation exchange extraction column to obtain a sample solution; the method comprises the following specific steps: accurately weighing 20g of apples, placing the apples in a centrifuge tube, accurately adding 50mL of a solution with a volume ratio of acetonitrile to water of 9:1, setting the frequency of 300 times/min to shake vigorously for 12min, standing the apples, placing the apples in a centrifuge, centrifuging the apples at 3000r/min for 5min, taking supernatant, continuously extracting residues with a solution with a volume ratio of acetonitrile to water of 25mL of 9:1, combining filtrates in a 100mL volumetric flask, adding 4mL of 1.0mol/L hydrochloric acid, fixing the volume with the solution with the volume ratio of acetonitrile to water of 9:1, and taking out 5mL of the filtrates for purification; activating the strong cation exchange extraction column by using 5% ammoniated methanol solution, pre-washing by using methanol and water in sequence, after 5mL of filtrate passes through the column, washing the strong cation exchange extraction column by using water and methanol in sequence, and draining; eluting the extraction column twice with 5% ammoniated methanol solution, drying the eluent at 40 deg.C with nitrogen, adding 1.00mL of mobile phase, vortex for 30s, and passing through 0.22 μm organic filter membrane for instrument measurement;
(2) preparing a standard solution of matrine, and preparing a plurality of standard series solutions according to concentration gradient; precisely weighing appropriate amount of matrine standard substance, dissolving and diluting with methanol, transferring to volumetric flask and fixing volume to obtain matrine standard stock solution with appropriate concentration, respectively sucking appropriate amount of stock solution, and making into serial standard working solutions with concentration of 0.1ug/ml, 0.3ug/ml, 0.5ug/ml, 1.0ug/ml, 5.0ug/ml and 10.0ug/ml by flow matching.
(3) Detecting and analyzing the sample solution and the standard solution by adopting a high performance liquid chromatography, and obtaining the content of the matrine by utilizing an external standard method, wherein the high performance liquid chromatography detection and analysis comprises the following steps: respectively injecting the standard working solution and the sample working solution into a high performance liquid chromatograph, performing high performance liquid chromatography analysis, quantitatively comparing the peak area of the standard working solution with the peak area of the sample working solution, retaining time for qualitative determination, calculating by an external standard method, and calculating the content of matrine in the sample; the method is characterized in that the chromatographic conditions of the high performance liquid chromatography are as follows:
a chromatographic column: c18 chromatographic column with column length of 150mm, inner diameter of 4.6mm and particle size of 5 μm; column temperature: 30 +/-5 ℃;
sample introduction amount: 10 mu L of the solution; the mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is methanol, the mobile phase B is a 5% disodium hydrogen phosphate aqueous solution with the pH value of 9, and the volume ratio of the mobile phase A to the mobile phase B is 40: 60; flow rate: 1.0 mL/min; detection wavelength: the wavelength was set at 215 nm.
TABLE 1 recovery and precision results of four samples of matrine
From the above table, it can be seen that the method of the present invention is used for detecting matrine in four products of cucumber, potato, cabbage and apple, the relative deviation is less than 9.8%, and the recovery rate is more than 80%. Therefore, the pretreatment method for detecting the matrine in the fruits and vegetables by the solid-phase extraction-high performance liquid chromatography is simple and easy to operate, high in extraction efficiency, good in purification effect, high in detection precision, sensitive in detection, short in detection time and high in detection efficiency. As can be seen from FIG. 1, the high performance liquid chromatography is used for detecting a target peak at 10.983min in a liquid chromatogram of the potato added with 1.0mg/kg matrine, and the detection is convenient and quick.
Claims (4)
1. A high performance liquid chromatography detection method for matrine pesticide residue in fruits and vegetables is characterized by comprising the following steps:
(1) extracting matrine in a sample to be detected by adopting a solution with the volume ratio of acetonitrile to water being 9:1, purifying, extracting and redissolving an obtained extracting solution by using a strong cation exchange extraction column to obtain a sample solution, and specifically comprising the following steps: accurately weighing 20g of a sample to be detected, placing the sample into a centrifuge tube, accurately adding 50mL of acetonitrile-water volume ratio 9:1 solution, setting the frequency of 250-; activating the strong cation exchange extraction column by using 5% ammoniated methanol solution, pre-washing by using methanol and water in sequence, after 5mL of filtrate passes through the column, washing the strong cation exchange extraction column by using water and methanol in sequence, and draining; eluting the extraction column twice with 5% ammoniated methanol solution, drying the eluent at 40 deg.C with nitrogen, adding 1.00mL of mobile phase, vortex for 30s, and passing through 0.22 μm organic filter membrane for instrument measurement;
(2) preparing a standard solution of matrine, and preparing a plurality of standard series solutions according to concentration gradient;
(3) detecting and analyzing the sample solution and the standard solution by high performance liquid chromatography, and obtaining the content of matrine by an external standard method.
2. The high performance liquid chromatography detection method for matrine pesticide residue in fruits and vegetables according to claim 1, characterized by comprising the following steps:
(1) extracting matrine in a sample to be detected by adopting a solution with the volume ratio of acetonitrile to water being 9:1, purifying, extracting and redissolving the obtained extracting solution by using a strong cation exchange extraction column to obtain a sample solution; the method comprises the following specific steps: accurately weighing 20g of a sample to be detected, placing the sample into a centrifuge tube, accurately adding 50mL of acetonitrile-water volume ratio 9:1 solution, setting the frequency of 300 times/min to shake vigorously for 12min, placing the sample into a centrifuge after standing, centrifuging the sample at 3000r/min for 5min, taking supernatant, continuously extracting residues with 25mL of acetonitrile-water volume ratio 9:1 solution, combining filtrates in a 100mL volumetric flask, adding 4mL of 1.0mol/L hydrochloric acid, performing constant volume with acetonitrile-water volume ratio 9:1 solution, and taking out 5mL of filtrate for purification; activating the strong cation exchange extraction column by using 5% ammoniated methanol solution, pre-washing by using methanol and water in sequence, after 5mL of filtrate passes through the column, washing the strong cation exchange extraction column by using water and methanol in sequence, and draining; eluting the extraction column twice with 5% ammoniated methanol solution, drying the eluent at 40 deg.C with nitrogen, adding 1.00mL of mobile phase, vortexing for 30s, and passing through 0.22 μm organic filter membrane for instrument determination.
(2) Preparing a standard solution of matrine, and preparing a plurality of standard series solutions according to concentration gradient;
(3) detecting and analyzing the sample solution and the standard solution by high performance liquid chromatography, and obtaining the content of matrine by an external standard method.
3. The high performance liquid chromatography detection method for matrine pesticide residue in fruits and vegetables according to claim 1 or 2, characterized in that the preparation of standard series solution comprises the following steps:
precisely weighing appropriate amount of matrine standard substance, dissolving and diluting with methanol, transferring to volumetric flask and fixing volume to obtain matrine standard stock solution with appropriate concentration, respectively sucking appropriate amount of stock solution, and making into serial standard working solutions with concentration of 0.1ug/ml, 0.3ug/ml, 0.5ug/ml, 1.0ug/ml, 5.0ug/ml and 10.0ug/ml by flow matching.
4. The high performance liquid chromatography detection method for matrine pesticide residue in fruits and vegetables according to claim 1 or 2, the high performance liquid chromatography detection analysis comprises the following steps: respectively injecting the standard working solution and the sample working solution into a high performance liquid chromatograph, performing high performance liquid chromatography analysis, quantitatively comparing the peak area of the standard working solution with the peak area of the sample working solution, retaining time for qualitative determination, calculating by an external standard method, and calculating the content of matrine in the sample; the method is characterized in that the chromatographic conditions of the high performance liquid chromatography are as follows:
a chromatographic column: c18 chromatographic column with column length of 150mm, inner diameter of 4.6mm and particle size of 5 μm; column temperature: 30 +/-5 ℃;
sample introduction amount: 10 mu L of the solution; the mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is methanol, the mobile phase B is a 5% disodium hydrogen phosphate aqueous solution with the pH value of 9, and the volume ratio of the mobile phase A to the mobile phase B is 40: 60; flow rate: 1.0 mL/min; detection wavelength: the wavelength was set at 215 nm.
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| CN114924015A (en) * | 2022-06-16 | 2022-08-19 | 中南大学 | Method for rapidly detecting matrine and oxymatrine |
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