CN112924579A - Seabed mud pretreatment method and method for detecting content of rimantadine and/or memantine in seabed mud - Google Patents

Seabed mud pretreatment method and method for detecting content of rimantadine and/or memantine in seabed mud Download PDF

Info

Publication number
CN112924579A
CN112924579A CN202110107427.1A CN202110107427A CN112924579A CN 112924579 A CN112924579 A CN 112924579A CN 202110107427 A CN202110107427 A CN 202110107427A CN 112924579 A CN112924579 A CN 112924579A
Authority
CN
China
Prior art keywords
column
rimantadine
memantine
solution
detected
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.)
Pending
Application number
CN202110107427.1A
Other languages
Chinese (zh)
Inventor
田秀慧
徐英江
张秀珍
韩典峰
任传博
宫向红
黄会
刘慧慧
薛敬林
张华威
罗晶晶
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shandong Marine Resource and Environment Research Institute
Original Assignee
Shandong Marine Resource and Environment Research Institute
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Shandong Marine Resource and Environment Research Institute filed Critical Shandong Marine Resource and Environment Research Institute
Priority to CN202110107427.1A priority Critical patent/CN112924579A/en
Publication of CN112924579A publication Critical patent/CN112924579A/en
Pending legal-status Critical Current

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
    • G01N30/26—Conditioning of the fluid carrier; Flow patterns
    • G01N30/28—Control of physical parameters of the fluid carrier
    • G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
    • 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
    • G01N30/26—Conditioning of the fluid carrier; Flow patterns
    • G01N30/28—Control of physical parameters of the fluid carrier
    • G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
    • 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
    • G01N30/62—Detectors specially adapted therefor
    • G01N30/72—Mass spectrometers
    • 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
    • G01N30/26—Conditioning of the fluid carrier; Flow patterns
    • G01N30/28—Control of physical parameters of the fluid carrier
    • G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
    • G01N2030/324—Control of physical parameters of the fluid carrier of pressure or speed speed, flow rate

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 belongs to the technical field of chemical analysis, and particularly relates to a method for pretreating seabed mud and a method for detecting the content of rimantadine and/or memantine in the seabed mud. The invention provides a pretreatment method of seabed mud, which comprises the following steps: mixing a seabed mud sample, internal standard working solution, carbon powder and N-propyl ethylenediamine to obtain suspension; the internal standard substance of the internal standard working solution is rimantadine hydrochloride-D4(ii) a Mixing a mixed solution of acetic acid and acetonitrile with the suspension for extraction to obtain an extracting solution; subjecting the extract toAnd (5) collecting the column to obtain the substance to be detected. According to the invention, the rimantadine and the memantine in the seabed sediment can be efficiently extracted through pretreatment, other impurities in the seabed sediment can be removed layer by layer, the concentration of the object to be detected is improved, and the accuracy of the detection result is improved.

Description

Seabed mud pretreatment method and method for detecting content of rimantadine and/or memantine in seabed mud
Technical Field
The invention belongs to the technical field of chemical analysis, and particularly relates to a method for pretreating seabed mud and a method for detecting the content of rimantadine and/or memantine in the seabed mud.
Background
Rimantadine and memantine are amantadine analogs, used for the prevention and treatment of influenza virus infection in animals, have superior clinical efficacy to amantadine, and have less adverse reactions than amantadine. The residual amount in the environment is more and more with the use of rimantadine and memantine. The research shows that rimantadine and memantine have influence on human health. However, because rimantadine and memantine are low in price and good in effect, farmers mix antiviral drugs such as rimantadine and memantine into the feed in order to avoid economic loss caused by death of livestock and poultry caused by virus infection. With the culture wastewater and the residues in the medical field entering the water body through various ways, the wastewater containing rimantadine and memantine flows into the sea, causing the pollution of the marine environment, and finally affecting the human health by accumulating in organisms such as algae, shellfish, fish and the like and migrating along with the food chain.
There are few studies on the detection methods of rimantadine and memantine, and the only few detection methods are also directed to the detection of drug residues in foods or feeds of animal origin, such as gas chromatography or liquid chromatography. However, there are few reports on the detection techniques of rimantadine and memantine in the bottom sediment.
Disclosure of Invention
In view of the above, the invention provides a method for pretreating subsea mud and a method for detecting the content of rimantadine and/or memantine in the subsea mud. The pretreatment method of the seabed mud provided by the invention can be used for better extracting and purifying the seabed mud extracting solution, and is favorable for obtaining an accurate detection result.
The invention provides a pretreatment method of seabed mud, which comprises the following steps:
mixing a seabed mud sample, internal standard working solution, carbon powder and N-propyl ethylenediamine to obtain suspension; the internal standard substance of the internal standard working solution is rimantadine hydrochloride-D4；
Mixing a mixed solution of acetic acid and acetonitrile with the suspension for extraction to obtain an extracting solution;
performing column collection on the extracting solution to obtain a substance to be detected; the column for collecting the column is an MCX column or an HLB column.
Preferably, the column further comprises before collection: collecting the column and activating the column;
the activating solvent is methanol and water, and the volume ratio of the methanol to the water is 0.8-1.2: 1.
Preferably, the column collection comprises the steps of:
passing the extracting solution through a column, and eluting to obtain an eluent;
removing the solvent from the eluent to obtain a substance to be detected;
when the column for collecting the column is an MCX column, the eluent for eluting is a mixed solution of ammonia water and methanol;
when the column for collecting the column is an HLB column, the step of passing the column further comprises leaching, the solvent for leaching is ultrapure water, and the eluent for elution is acetonitrile.
Preferably, the carbon powder comprises C18 powder or C8 powder;
the particle size of the carbon powder is 40-100 mu m.
Preferably, the mass ratio of the seabed mud sample to the carbon powder to the N-propyl ethylenediamine is 1: 2.8-3.2, and the mass of the seabed mud sample is calculated by dry weight.
Preferably, the volume ratio of acetic acid to acetonitrile in the mixed solution of acetic acid and acetonitrile is 0.8-1.2: 98.8-99.2.
Preferably, the extraction is performed under ultrasonic conditions; the power of the ultrasonic wave is 1000-1500W, and the time is 25-35 min.
The invention also provides a method for detecting the content of rimantadine and/or memantine in the seabed mud, which comprises the following steps:
mixing a substance to be detected and an acetonitrile-ammonium acetate solution of formic acid to obtain a liquid to be detected;
performing liquid chromatography tandem mass spectrometry detection on the liquid to be detected, and substituting the ratio of the peak areas of the rimantadine and the memantine to the peak area of the internal standard substance into a preset rimantadine standard curve or memantine standard curve respectively to obtain the concentration of the rimantadine and/or the memantine in the liquid to be detected; the object to be detected is prepared according to the pretreatment method of the technical scheme.
Preferably, the liquid chromatography conditions comprise: a chromatographic column: c18 column, 2.1mm × 100mm, filler particle size 1.9 μm; mobile phase A: methanol; mobile phase B: the volume concentration of the formic acid aqueous solution is 0.1%.
Preferably, the mass spectrometry conditions comprise: an ion source: electrospray ionization; the detection method comprises the following steps: a multiple reaction monitoring mode; capillary voltage is 3500V; the ion source temperature is 350 ℃; shell gas (nitrogen gas) 40; auxiliary gas (nitrogen gas) 12; the temperature of the ion transfer tube is 350 ℃.
The invention provides a pretreatment method of seabed mud, which comprises the following steps: mixing a seabed mud sample, an internal standard working solution, carbon powder and N-CMixing ethylene diamine to obtain suspension; the internal standard substance of the internal standard working solution is rimantadine hydrochloride-D4(ii) a Mixing a mixed solution of acetic acid and acetonitrile with the suspension for extraction to obtain an extracting solution; performing column collection on the extracting solution to obtain a substance to be detected; the column for collecting the column is an MCX column or an HLB column. In the invention, the carbon powder can adsorb organic matters in the seabed mud, and the N-propyl ethylenediamine can also play a role in purifying the extracting solution, thereby avoiding the influence of coexisting substances on the accuracy of the detection result. According to the invention, the loss amount of the object to be detected in the pretreatment process can be accurately known by adding the internal standard working solution, so that the accuracy of the detection result is improved. The invention adopts the mixed solution of acetic acid and acetonitrile for extraction, and can fully extract rimantadine and memantine in the seabed sediment. The invention can improve the purity of the object to be detected and the accuracy of the detection result by purifying the extracting solution by using the MCX column or the HLB column.
Drawings
FIG. 1 is a selected ion flow graph of a standard solution having a mass concentration of rimantadine, memantine, and an internal standard of 10.0ng/mL, respectively;
FIG. 2 is a selected ion flow diagram of a test solution prepared in comparative example 2;
FIG. 3 is a selected particle flow diagram of the resulting detection solutions for rimantadine, memantine, and internal standard at a concentration of 10.0 μ g/kg, wherein the detection solutions for rimantadine, memantine, and internal standard at a concentration of 10.0 μ g/kg are obtained by mixing the solution to be detected, the rimantadine standard solution, and the memantine standard solution prepared in comparative example 2.
Detailed Description
The invention provides a pretreatment method of seabed mud, which comprises the following steps:
mixing a seabed mud sample, internal standard working solution, carbon powder and N-propyl ethylenediamine to obtain suspension; the internal standard substance of the internal standard working solution is rimantadine hydrochloride-D4；
Mixing a mixed solution of acetic acid and acetonitrile with the suspension for extraction to obtain an extracting solution;
performing column collection on the extracting solution to obtain a substance to be detected; the column for collecting the column is an MCX column or an HLB column.
Mixing a seabed mud sample, internal standard working solution, carbon powder and N-propyl ethylenediamine to obtain suspension; the internal standard substance of the internal standard working solution is rimantadine hydrochloride-D4. In the invention, the mass concentration of the internal standard working solution is preferably 95-105 mug/L, and more preferably 100 mug/L. In the invention, the water content of the seabed mud sample is preferably 20-30%, and more preferably 25-28%. The source of the bottom sediment sample is not particularly limited, and in the embodiment of the invention, the bottom sediment sample is bottom sediment in a yellow sea area or bottom sediment in a Bohai sea area. In the present invention, the ratio of the mass of the sediment sample to the volume of the internal standard working solution is preferably 1 g: 90-110 μ L, more preferably 1 g: 95-100 μ L. In the present invention, the carbon powder preferably includes C18 powder or C8 powder, and more preferably C18 powder. In the present invention, the particle size of the carbon powder is preferably 40 to 100 μm, and more preferably 40 to 60 μm. In the invention, the mass ratio of the seabed mud sample, the carbon powder and the N-propylethylenediamine is preferably 1: 2.8-3.2, and more preferably 1:3: 3. In the present invention, the mass of the seabed mud sample is preferably calculated as dry weight. The mixing is not particularly limited in the present invention as long as it can be mixed uniformly. In the invention, the carbon powder can adsorb organic matters in the seabed mud, and the N-propyl ethylenediamine can also play a role in purifying the extracting solution, thereby avoiding the influence of coexisting substances on the accuracy of the detection result.
After the suspension is obtained, the mixed solution of acetic acid and acetonitrile is mixed with the suspension for extraction to obtain the extracting solution. In the invention, the volume ratio of acetic acid to acetonitrile in the mixed solution of acetic acid and acetonitrile is preferably 0.8-1.2: 98.8-99.2, and more preferably 1: 99; in the invention, the acetic acid in the mixed solution of the acetic acid and the acetonitrile provides an acidic environment, the solubility of the acetonitrile to the rimantadine and/or memantine is improved, and the extraction rate is improved. In the invention, the volume of the mixed solution of acetic acid and acetonitrile and the mass ratio of the seabed sediment sample are preferably 20-40 mL: 1g, more preferably 20-25 mL: 1g of the total weight of the composition. In the invention, the extraction is preferably carried out under the condition of ultrasound, and the power of the ultrasound is preferably 1000-1500W, and more preferably 1000-1200W; the time is preferably 25 to 35min, and more preferably 30 to 33 min.
In the present invention, the extraction preferably further comprises centrifugation to obtain an extract; the rotation speed of the centrifugation is preferably 6500-7500 r/min, more preferably 6800-7000 r/min; the time is preferably 8 to 15min, and more preferably 10 to 12 min.
After the extracting solution is obtained, carrying out column collection on the extracting solution to obtain a substance to be detected; the column for collecting the column is an MCX column or an HLB column. In the present invention, the column before collection preferably further comprises: the column was collected and activated with a column. In the invention, the activating solvent is preferably methanol and water, and the volume ratio of the methanol to the water is preferably 0.8-1.2: 1, and more preferably 1: 1. In the invention, the dosage of the methanol is preferably 1-2 times of the column volume, and more preferably 1.2-1.5 times of the column volume; the amount of the water is preferably 1-2 times of the column volume, and more preferably 1.2-1.5 times of the column volume.
In the present invention, the column collection preferably comprises the steps of:
passing the extracting solution through a column, and eluting to obtain an eluent;
and removing the solvent from the eluent to obtain the substance to be detected.
The invention passes the extracting solution through the column, and then elutes to obtain the eluent. In the invention, when the column for collecting the column is an MCX column, the eluent for elution is preferably a mixed solution of ammonia water and methanol, and the volume concentration of the ammonia water is preferably 0.8-1.2%, and more preferably 0.9-1.1%; the volume ratio of the ammonia water to the methanol is 0.8-1.2: 99, and more preferably 1: 99. In the invention, the amount of the eluent is preferably 0.6-1.2 times of the column volume, and more preferably 0.8-1.0 times of the column volume.
In the present invention, when the column for column collection is an HLB column, it is preferable to further include, before passing through the column: and (4) carrying out rotary evaporation on the extracting solution, and dissolving the solid obtained by rotary evaporation in water to obtain a solution to be subjected to column chromatography. In the invention, the mass ratio of the solid to the water is preferably 4-8: 1, more preferably 5 to 6: 1.
in the invention, the HLB column preferably comprises leaching after passing through the column, the leaching solvent is preferably ultrapure water, and the dosage of the leaching solvent is preferably 0.5-2 times of the column volume, and more preferably 0.5-1 times of the column volume; the eluent for elution is preferably acetonitrile; the dosage of the eluent is preferably 0.6-1.2 times of the column volume, and more preferably 0.8-1.0 times of the column volume.
After the eluent is obtained, the eluent is subjected to solvent removal to obtain the object to be detected. In the invention, the desolvation is preferably carried out under the condition of nitrogen purging, and the temperature of the nitrogen is preferably 40-48 ℃, and more preferably 43-45 ℃. In the present invention, the time for purging is not particularly limited as long as the solvent can be completely removed.
The invention selects a specific column and an eluant in the column collection process, can improve the purity of the object to be detected, and is beneficial to improving the accuracy of the detection result.
The invention also provides a method for detecting the content of rimantadine and/or memantine in the seabed mud, which comprises the following steps:
mixing a substance to be detected and an acetonitrile-ammonium acetate solution of formic acid to obtain a liquid to be detected;
performing liquid chromatography tandem mass spectrometry detection on the liquid to be detected, and substituting the obtained ratio of the peak areas of rimantadine and memantine to the peak area of the internal standard substance into a preset rimantadine standard curve or memantine standard curve respectively to obtain the concentration of the rimantadine and/or memantine in the liquid to be detected; the object to be detected is prepared according to the pretreatment method of the technical scheme.
The method comprises the step of mixing a substance to be detected and an acetonitrile-ammonium acetate solution of formic acid to obtain a liquid to be detected. In the invention, the mass percentage of formic acid in the acetonitrile-ammonium acetate solution of formic acid is preferably 0.08-0.12%, and more preferably 0.09-0.1%; the mass ratio of acetonitrile to ammonium acetate in the acetonitrile-ammonium acetate solution of formic acid is preferably 93-98: 5, and more preferably 95-96: 5.
After the liquid to be detected is obtained, performing liquid chromatography tandem mass spectrometry detection on the liquid to be detected, and substituting the ratio of the peak areas of the rimantadine and memantine to the peak area of the internal standard substance into a preset standard curve to obtain the concentration of the rimantadine and/or memantine in the liquid to be detected; the liquid to be detected is prepared according to the pretreatment method of the technical scheme. In the present invention, the liquid chromatography conditions include: a chromatographic column: c18 column, 2.1mm × 100mm, filler particle size 1.9 μm; mobile phase A: methanol; mobile phase B: the volume concentration of the formic acid aqueous solution is 0.1 percent; in the present invention, a gradient elution is preferably adopted in the liquid chromatography detection process, the flow rate of the elution is preferably 0.25mL/min, and the procedure of the gradient elution is shown in table 1:
table 1 liquid chromatography mobile phase gradient elution procedure
Time (min) Flow rate (mL/min) Mobile phase A (%) Mobile phase B (%)
0 0.25 5 95
0.5 0.25 5 95
3 0.25 95 5
5 0.25 95 5
6 0.25 5 95
7 0.25 5 95
In the present invention, the mass spectrometry conditions include: an ion source: electrospray ionization; the detection method comprises the following steps: a multiple reaction monitoring mode; capillary voltage is 3500V; the ion source temperature is 350 ℃; shell gas (nitrogen gas) 40; auxiliary gas (nitrogen gas) 12; the temperature of the ion transfer tube is 350 ℃.
In the present invention, the standard curve is preferably obtained as follows:
respectively mixing the rimantadine standard solution and the memantine standard solution with the internal standard working solution to respectively obtain rimantadine standard detection solution and memantine standard detection solution;
respectively carrying out liquid chromatography tandem mass spectrometry detection on the rimantadine standard detection solution and the memantine standard detection solution to obtain the ratio of the peak area of rimantadine and/or memantine to the peak area of the internal standard substance;
and respectively carrying out linear fitting on the ratio of the peak area of the rimantadine to the peak area of the internal standard substance, the concentration of the rimantadine standard detection liquid, the ratio of the peak area of the memantine to the peak area of the internal standard substance and the concentration of the memantine standard detection liquid to respectively obtain a rimantadine standard curve and a memantine standard curve.
In the present invention, the preparation method of the rimantadine standard test solution preferably includes:
mixing 10 mu L, 20 mu L, 40 mu L, 50 mu L and 100 mu L of rimantadine standard use solution with the mass concentration of 1.0mg/L and 100 mu L of internal standard working solution with the mass concentration of 100 mu g/L respectively, and then using distilled water to fix the volume to 1.00mL to obtain rimantadine standard detection solutions with the mass concentrations of 10 mu g/L, 20 mu g/L, 40 mu g/L, 50 mu g/L and 100 mu g/L respectively;
according to the technical scheme, the standard rimantadine use solution is replaced by the standard memantine use solution to obtain a standard memantine detection solution;
and respectively detecting the rimantadine standard detection liquid or the memantine standard detection liquid with different mass concentrations by using a liquid chromatography tandem mass spectrum, and drawing a standard curve by taking the ratio of the peak area of a solute in the standard detection liquid to the peak area of an internal standard substance as a vertical coordinate and the concentration of the standard detection liquid as a horizontal coordinate.
In the present invention, the standard curve of rimantadine is 0.138589+0.234309x, R2The linear range is 10-100 mu g/L and is 0.9994; the standard curve of memantine is 0.0466678+0.220968 x; r20.9969, and the linear range is 10-100 mu g/L. The standard curve obtained by the invention has good linear relation, which shows that the detection method provided by the invention can be used for accurately and quantitatively detecting rimantadine and memantine.
Obtaining the ratio of the peak area of the sample to be detected in the sample to be detected to the peak area of the internal standard substance, substituting the ratio into the standard curve to respectively obtain the concentration of rimantadine or memantine
In order to further illustrate the present invention, the following embodiments are described in detail, but they should not be construed as limiting the scope of the present invention.
Example 1
Mixing 10 mu L, 20 mu L, 40 mu L, 50 mu L and 100 mu L of rimantadine standard use solution with the mass concentration of 1.0mg/L and 100 mu L of internal standard working solution with the mass concentration of 100 mu g/L respectively, and then using distilled water to fix the volume to 1.00mL to obtain rimantadine standard detection solutions with the mass concentrations of 10 mu g/L, 20 mu g/L, 40 mu g/L, 50 mu g/L and 100 mu g/L respectively;
preparing a memantine standard detection solution according to the method;
and respectively detecting the rimantadine standard detection solution or the memantine standard detection solution with different mass concentrations by using a liquid chromatography tandem mass spectrum, and drawing a standard curve by taking the ratio of the peak area of a solute in the standard detection solution to the peak area of an internal standard substance as a vertical coordinate and the concentration of the standard solution as a horizontal coordinate.
The standard curve of rimantadine is 0.138589+0.234309x, R2The linear range is 10-100 mu g/L and is 0.9994; the standard curve of memantine is that y is 0.0466678+0.220968x, R20.9969, and the linear range is 10-100 mu g/L.
Example 2
Mixing 1g of a sea sediment (taken from Bohai sea area) sample with the water content of 25%, 100 mu L of internal standard working solution with the mass concentration of 100 mu g/L, 3g of C18 powder with the average particle size of 40 mu m and 3g of N-propyl ethylenediamine to obtain a suspension;
mixing the suspension with 25mL of mixed solution of acetic acid and acetonitrile (the mass concentration of acetic acid is x%, and the volume ratio of acetic acid to acetonitrile is 1:99), performing ultrasonic treatment at 1000W for 30min, centrifuging at 7000r/min for 10min, and collecting supernatant to obtain extractive solution;
activating the MCX column by using 10mL of methanol and 10mL of water to obtain an activated MCX column; the dosage of the methanol and the water is independently 2 times of the column volume;
and (3) passing the extracting solution through an activated MCX column, eluting by using 10mL of mixed solution of ammonia water and methanol with the volume concentration of 1% (the volume ratio of the ammonia water to the methanol is 1:99), and drying the collected eluent by nitrogen at 45 ℃ to obtain the substance to be detected.
Example 3
Mixing 1g of a sea sediment (taken from Bohai sea area) sample with the water content of 25%, 100 mu L of internal standard working solution with the mass concentration of 100 mu g/L, 3g of C18 powder with the average particle size of 40 mu m and 3g of N-propyl ethylenediamine to obtain a suspension;
mixing the suspension with 25mL of mixed solution of acetic acid and acetonitrile (the volume ratio of acetic acid to acetonitrile is 1:99), performing ultrasonic treatment at 1000W for 30min, centrifuging at the rotating speed of 7000r/min for 10min, performing rotary evaporation on the centrifuged supernatant, and dissolving the solid obtained by rotary evaporation in 5mL of ultrapure water to obtain an extracting solution;
activating the HLB column by using 10mL of methanol and 10mL of water to obtain an activated HLB column; the dosage of the methanol and the water is independently 2 times of the column volume;
and (3) passing the extracting solution through an activated HLB (hydrophile-lipophile balance) column, eluting by using 3mL of ultrapure water and 5mL of acetonitrile in sequence, and drying the collected eluent by using nitrogen at 45 ℃ to obtain the substance to be detected.
Comparative example 1
Pretreating a subsea mud sample according to example 1, except that the subsea mud is obtained from the gulf of leizhou, and the subsea mud sample does not contain rimantadine and memantine; the selected purifying column is an MAX column.
Comparative example 2
A subsea mud sample was pretreated as in example 1, except that the subsea mud was obtained from the gulf of lazhou, and the subsea mud sample did not contain rimantadine and memantine.
Comparative example 3
The subsea mud sample was pretreated according to the method of comparative example 1, except that the purification column used was an HBL column
Example 4
Dissolving the to-be-detected substances prepared in the examples 2 and 3 and the comparative example 2 in 1mL of acetonitrile ammonium acetate solution of formic acid (the mass percentage of the formic acid is 0.1 percent, and the mass ratio of acetonitrile to ammonium acetate is 95: 5) to obtain a to-be-detected solution;
and respectively carrying out liquid chromatography tandem mass spectrometry detection on the obtained liquid to be detected, respectively substituting the ratio of the peak areas of the rimantadine and the memantine to the peak area of the internal standard substance into the standard curve to obtain the concentrations of the rimantadine and the memantine in the liquid to be detected, and listing the results in table 2.
The liquid chromatography conditions were: a chromatographic column: c18 column, 2.1mm × 100mm, particle size 1.9 μm; mobile phase A: methanol; mobile phase B: 0.1% formic acid in water; the procedure for gradient elution is shown in table 1:
the mass spectrum conditions are as follows: an ion source: electrospray ionization; the detection method comprises the following steps: a multiple reaction monitoring mode; capillary voltage is 3500V; the ion source temperature is 350 ℃; shell gas (nitrogen gas) 40; auxiliary gas (nitrogen gas) 12; the temperature of the ion transfer tube is 350 ℃.
TABLE 2 test results of specimens obtained in examples 2 and 3 and comparative example 2
Examples Rimantadine concentration (μ g/kg) Memantine concentration (μ g/kg)
Example 2 5.15 8.03
Example 3 6.03 6.25
Comparative example 2 Not detected out Not detected out
Performing liquid chromatography tandem mass spectrometry on the standard solutions with the mass concentrations of rimantadine, memantine and the internal standard substance of 10.0ng/mL respectively to obtain a selected ion flow diagram, which is shown in figure 1. Obtaining the rimantadine, the memantine and the rimantadine hydrochloride-D according to the detection result4The parent ion and daughter ion collision energies of (a) are specifically shown in Table 3.
TABLE 3 rimantadine, memantine and rimantadine hydrochloride-D4Selection ofIon flow summary table
Figure BDA0002918086300000101
And (3) performing liquid chromatography tandem mass spectrometry detection on the solution to be detected prepared in the comparative example 2 to obtain a selected ion flow diagram, which is shown in figure 2.
And (3) mixing the solution to be detected, the rimantadine standard solution and the memantine standard solution which are prepared in the comparative example 2 to obtain detection solutions with the concentration of the rimantadine, the memantine and the internal standard substance of 10.0 mug/kg, and performing liquid chromatography tandem mass spectrometry to obtain a selected ion flow diagram, as shown in figure 3.
With reference to fig. 1 to 3 and table 3, it can be seen that the comparative example 2 does not contain rimantadine and memantine, the detection result is 0, and the detection result of the liquid to be detected after the sample to be detected in the comparative example 2 is mixed with the standard liquid is consistent with the detection result of the standard liquid, which indicates that the method of the present invention can detect rimantadine and memantine and can well detect the content of rimantadine and memantine in the seafloor mud.
Respectively adding rimantadine and memantine into the to-be-detected liquid prepared in the comparative example 1 and the comparative example 2 to obtain to-be-detected liquids with the mass concentrations of 10 mug/kg, 50 mug/kg and 100 mug/kg independently; respectively carrying out liquid chromatography tandem mass spectrometry detection on the solutions to be detected with different mass concentrations, and carrying out seven detections on each solution to be detected with each mass concentration, wherein the detection results are listed in table 4.
TABLE 4 detection of the recovery of rimantadine and memantine by liquid chromatography tandem mass spectrometry
Figure BDA0002918086300000102
As can be seen from the results in Table 4, the detection results obtained by different columns for pretreatment are different, and the detection method provided by the invention has higher accuracy in detecting rimantadine and memantine.
Although the present invention has been described in detail with reference to the above embodiments, it is only a part of the embodiments of the present invention, not all of the embodiments, and other embodiments can be obtained without inventive step according to the embodiments, and the embodiments are within the scope of the present invention.

Claims (10)

1. A pretreatment method of seabed mud comprises the following steps:
mixing a seabed mud sample, internal standard working solution, carbon powder and N-propyl ethylenediamine to obtain suspension; the internal standard substance of the internal standard working solution is rimantadine hydrochloride-D4；
Mixing a mixed solution of acetic acid and acetonitrile with the suspension for extraction to obtain an extracting solution;
performing column collection on the extracting solution to obtain a substance to be detected; the column for collecting the column is an MCX column or an HLB column.
2. The method of pretreating seafloor mud of claim 1, further comprising, prior to the collecting, by the column: collecting the column and activating the column;
the activating solvent is methanol and water, and the volume ratio of the methanol to the water is 0.8-1.2: 1.
3. The method for the pretreatment of bottom mud according to claim 1 or 2, wherein the column collection comprises the steps of:
passing the extracting solution through a column, and eluting to obtain an eluent;
removing the solvent from the eluent to obtain a substance to be detected;
when the column for collecting the column is an MCX column, the eluent for eluting is a mixed solution of ammonia water and methanol;
when the column for collecting the column is an HLB column, the step of passing the column further comprises leaching, the solvent for leaching is ultrapure water, and the eluent for elution is acetonitrile.
4. The method for pretreating seabed mud according to claim 1, wherein the carbon powder comprises C18 powder or C8 powder;
the particle size of the carbon powder is 40-100 mu m.
5. The method for pretreating seabed mud according to claim 1 or 4, wherein the mass ratio of the seabed mud sample, the carbon powder and the N-propyl ethylene diamine is 1: 2.8-3.2, and the mass of the seabed mud sample is calculated by dry weight.
6. The method for pretreating seabed mud according to claim 1, wherein the volume ratio of acetic acid to acetonitrile in the mixed solution of acetic acid and acetonitrile is 0.8-1.2: 98.8-99.2.
7. The method for the pretreatment of bottom sediment according to claim 1 or 6, wherein the extraction is performed under ultrasonic conditions; the power of the ultrasonic wave is 1000-1500W, and the time is 25-35 min.
8. A method for detecting the content of rimantadine and/or memantine in seabed mud comprises the following steps:
mixing a substance to be detected and an acetonitrile-ammonium acetate solution of formic acid to obtain a liquid to be detected;
performing liquid chromatography tandem mass spectrometry detection on the liquid to be detected, and substituting the ratio of the peak areas of the rimantadine and the memantine to the peak area of the internal standard substance into a preset rimantadine standard curve or memantine standard curve respectively to obtain the concentration of the rimantadine and/or the memantine in the liquid to be detected; the object to be tested is prepared according to the pretreatment method of any one of claims 1 to 7.
9. The detection method according to claim 8, wherein the liquid chromatography conditions comprise: a chromatographic column: c18 column, 2.1mm × 100mm, filler particle size 1.9 μm; mobile phase A: methanol; mobile phase B: the volume concentration of the formic acid aqueous solution is 0.1%.
10. The detection method of claim 8, wherein the mass spectrometry conditions comprise: an ion source: electrospray ionization; the detection method comprises the following steps: a multiple reaction monitoring mode; capillary voltage is 3500V; the ion source temperature is 350 ℃; shell gas (nitrogen gas) 40; auxiliary gas (nitrogen gas) 12; the temperature of the ion transfer tube is 350 ℃.
CN202110107427.1A 2021-01-27 2021-01-27 Seabed mud pretreatment method and method for detecting content of rimantadine and/or memantine in seabed mud Pending CN112924579A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202110107427.1A CN112924579A (en) 2021-01-27 2021-01-27 Seabed mud pretreatment method and method for detecting content of rimantadine and/or memantine in seabed mud

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202110107427.1A CN112924579A (en) 2021-01-27 2021-01-27 Seabed mud pretreatment method and method for detecting content of rimantadine and/or memantine in seabed mud

Publications (1)

Publication Number Publication Date
CN112924579A true CN112924579A (en) 2021-06-08

Family

ID=76166627

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202110107427.1A Pending CN112924579A (en) 2021-01-27 2021-01-27 Seabed mud pretreatment method and method for detecting content of rimantadine and/or memantine in seabed mud

Country Status (1)

Country Link
CN (1) CN112924579A (en)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20110110871A1 (en) * 2009-11-09 2011-05-12 Conopco, Inc., D/B/A Unilever Substituted 3-(phenoxymethyl) benzyl amines and personal care compositions
CN105628826A (en) * 2016-03-30 2016-06-01 山东省海洋资源与环境研究院 Ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) used for detecting residual quantity of adamantanamine in seawater and ocean sediments
CN110426462A (en) * 2019-07-03 2019-11-08 佛山科学技术学院 Amantadine, Rimantadine and the remaining method of Memantine in a kind of detection animal derived food

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20110110871A1 (en) * 2009-11-09 2011-05-12 Conopco, Inc., D/B/A Unilever Substituted 3-(phenoxymethyl) benzyl amines and personal care compositions
CN105628826A (en) * 2016-03-30 2016-06-01 山东省海洋资源与环境研究院 Ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) used for detecting residual quantity of adamantanamine in seawater and ocean sediments
CN110426462A (en) * 2019-07-03 2019-11-08 佛山科学技术学院 Amantadine, Rimantadine and the remaining method of Memantine in a kind of detection animal derived food

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
XU YJ 等: "Analysis of amantadine in Laminaria Japonica and seawater of Daqin Island by ultra high performance liquid chromatography with positive electrospray ionization tandem mass spectrometry", 《JOURNAL OF CHROMATOGRAPHY B》 *
吴银良 等: "同位素稀释液相色谱-串联质谱法测定鸡蛋中金刚烷胺类药物残留量", 《分析测试学报》 *
周剑 等: "两种样品处理方法对LC-MS/MS测定鸡肉中金刚烷胺基质效应比较", 《分析科学学报》 *

Similar Documents

Publication Publication Date Title
CN100346162C (en) Method for detecting citrinin content in red koji fermentation product
CN106153770B (en) A kind of Solid Phase Extraction of aquatic products glyphosate-liquid chromatography-mass spectrography detection method
CN105334277B (en) Quick pretreatment method and detection method for grape wine farming residual analysis
CN107216422A (en) A kind of preparation method and application of aflatoxin Restrict access material molecular engram separating medium
CN104931597A (en) Method capable of simultaneously detecting varieties of pesticide residues in aquatic product
CN105203666B (en) A kind of method of methyl mercury, ethyl mercury and inorganic mercury in detection aquatic feeds
CN109061016A (en) A kind of preparation method and application of the solid-phase extraction column of enriched biological amine
CN105044262B (en) Water polychlorinated biphenyl dispersive solid-phase extraction gas chromatography detection method
Liang et al. Comparative studies on the pH dependence of D OW of microcystin-RR and-LR using LC-MS
CN105954376B (en) It is a kind of that piperazine residual and the method for derivatization are extracted from fowl and porcine tissue
CN1187355C (en) Method for refining high-purity tetradoxin
Jimenez-Carmona et al. Ion-pair—supercritical fluid extraction of clenbuterol from food samples
CN112924579A (en) Seabed mud pretreatment method and method for detecting content of rimantadine and/or memantine in seabed mud
CN114235798B (en) Method for detecting putrefactive meat used in pet food
CN102393430B (en) Pretreatment kit for detecting Tilmicosin in animal muscle tissue and method thereof
CN107202840A (en) A kind of method for detecting 4 kinds of barbiturates downerns of fresh milk
CN105277631A (en) High performance liquid chromatography for detection of residual ceftiofur in pork tissues
CN110465269A (en) A kind of preparation method and applications of molecular engram metal organic framework composite material
CN116183773A (en) Method for rapidly determining flavomycin A in animal-derived food by utilizing liquid chromatography-mass spectrometry
CN105651890B (en) A kind of quick determination method of biogenic amine in aquatic product
CN111707763A (en) Method for rapidly detecting chloramphenicol in food
CN110412170B (en) Online solid phase extraction gas chromatography-mass spectrometry detection method for organotin in marine products
CN108760956B (en) Pretreatment device and content determination method for fluoroquinolone veterinary drug antibiotic sample in breeding wastewater
CN110412156B (en) Method for determining polychlorinated biphenyl in soil
CN111257439A (en) Method for detecting hydroxyl polybrominated diphenyl ethers in aquatic products by solid-phase extraction-ultra-high performance liquid chromatography tandem 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
RJ01 Rejection of invention patent application after publication
RJ01 Rejection of invention patent application after publication

Application publication date: 20210608