CN107102075A - A kind of method of illegal addition pappy shell in quick discriminating herbal tea - Google Patents
A kind of method of illegal addition pappy shell in quick discriminating herbal tea Download PDFInfo
- Publication number
- CN107102075A CN107102075A CN201710259454.4A CN201710259454A CN107102075A CN 107102075 A CN107102075 A CN 107102075A CN 201710259454 A CN201710259454 A CN 201710259454A CN 107102075 A CN107102075 A CN 107102075A
- Authority
- CN
- China
- Prior art keywords
- herbal tea
- pappy shell
- ammonium acetate
- solution
- sample
- 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.)
- Granted
Links
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
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)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention discloses a kind of method of illegal addition pappy shell in quick discriminating herbal tea, comprise the following steps：(1) liquid herbal tea sample is taken, it is ultrasonically treated, herbal tea sample is obtained, it is adjusted in alkalescent, organic solvent is added and sodium chloride is vortexed and extracted, centrifuge, layering adds ammonium acetate solution in supernatant, mixes, obtains prepare liquid；Or solid herbal tea sample is chosen, and organic solvent is added, it is ultrasonically treated, extract solution is cooled to room temperature, constant volume takes extract solution to add ammonium acetate solution, mixes, obtain prepare liquid；(2) prepare liquid is filtered through organic phase filter membrane, mobile phase is used as using acetic acid methanol solution and ammonium acetate solution, inverted chromatogram post separation, five kinds of main component morphines, codeine, thebaine, papaverine and coscopin in pappy shell are detected by eluent with ultra performance liquid chromatography tandem mass spectrometer.This method pre-treatment is simple, quick, qualitative, quantitative is accurate, can carry out quick, reliable detection to illegal addition opium poppy shell component in herbal tea.
Description
Technical field
The invention belongs to pappy shell identification technology field, and in particular to illegal addition pappy shell in a kind of quick discriminating herbal tea
Method.
Background technology
Pappy shell is commonly called as opium shell, with certain medical value, available for relieving pain, calm and cough-relieving.Its main component
For alkaloids such as morphine, codeine, thebaine, papaverine and coscopins.Illegal addition pappy shell, can increase cough-relieving effect in herbal tea
Really, but long-term consumption tool it is additive, can also patient is had the fidgets during large usage quantity, mental excitation, faint from fear situations such as.Small-mouthed jar
Grain shell belongs to official drug, is the control thing that national formal announcement is included in arcotic list, forbids being used for food, country defends meter
Committee, the Ministry of Public Security, National Drug Administration's relevant laws and regulations regulation are forbidden to use pappy shell in food, but some criminals
In order to strengthen cough-relieving effect of herbal tea, attract more customers, obtain bigger profit.Drink illegal addition pappy shell powder or
The herbal tea for the liquid that pappy shell infusion is obtained, easily makes not in contact with crossing drugs or having people's habituation of hypersensitivity to drugs, seriously
Endanger the health of consumer.
Not yet there are the national standard and pertinent literature of illegal addition pappy shell in quick discriminating herbal tea at present, one kind need to be set up
Reliable detection method.
The content of the invention
It is an object of the invention to provide a kind of method of illegal addition pappy shell in quick discriminating herbal tea, locate before this method
Reason is quick, can qualitative, quantitative differentiate the main component morphine of pappy shell in herbal tea sample, codeine, thebaine, papaverine and
Coscopin.
The above-mentioned purpose of the present invention is achieved through the following technical solutions：Illegally added in a kind of quick discriminating herbal tea
The method of pappy shell, comprises the following steps：
(1) liquid herbal tea sample is chosen, progress is ultrasonically treated, pappy shell main component in herbal tea sample is fully dissolved,
Room temperature is cooled to, herbal tea sample is obtained, herbal tea sample is taken, it is adjusted in alkalescent, organic solvent is added and sodium chloride carries out whirlpool
Rotation extraction, is then centrifuged again, is layered, is taken supernatant, ammonium acetate solution is then added in supernatant, is mixed, and prepare liquid is made；
Or choose solid herbal tea sample, add organic solvent, carry out it is ultrasonically treated, make in herbal tea sample pappy shell main into
Divide and be fully extracted, obtain extract solution, extract solution is cooled to room temperature, constant volume takes part extract solution addition ammonium acetate after constant volume molten
Liquid, is mixed, and prepare liquid is made；
(2) after prepare liquid is filtered through organic phase filter membrane, using acetate-methanol solution and ammonium acetate solution as mobile phase,
It is after inverted chromatogram post separation, eluent is main using five kinds in ultra performance liquid chromatography-tandem mass spectrometer detection pappy shell
Composition morphine, codeine, thebaine, papaverine and coscopin.
In above-mentioned quick discriminating herbal tea in the method for illegal addition pappy shell：
Liquid herbal tea sample in step (1) of the present invention can be bulk liquids herbal tea sample on the market.
When ultrasonically treated in step (1) of the present invention, preferably adjust temperature be 40~50 DEG C with 40khz frequency ultrasounds 15~
25min。
Ultrasonic extraction can be such that pappy shell active ingredient in herbal tea fully dissolves to herbal tea sample in a heated condition.
As one kind of the invention preferred embodiment, the equipment used when of the invention ultrasonically treated can be ultrasonic wave
The clean supersonic wave cleaning machine PL-S100 of cleaning machine, such as health scholar.
Herbal tea sample is taken in step (1) of the present invention, ammonia spirit or sodium hydroxide solution that pH value is 8~9 is preferably added to
It is adjusted in alkalescent, wherein herbal tea sample and ammonia spirit that pH value is 8~9 or sodium hydroxide solution is preferred with magnitude relation
For 1g：1mL~2mL；Further, herbal tea sample and pH value use magnitude relation for 8~9 ammonia spirit or sodium hydroxide solution
More preferably 1g：1mL.
The pH value of wherein regulation herbal tea sample can make the composition to be measured in herbal tea in pappy shell by ionic state for alkalescent
It is changed into molecular state, then the higher rate of recovery can be obtained with organic solvent such as methanol or acetonitrile extraction, while in ultra high efficiency liquid phase
There are preferable separating degree and detection sensitivity in chromatogram-tandem mass spectrum detection.
Organic solvent described in step (1) of the present invention and the herbal tea sample are preferably 2~4mL with magnitude relation：1g, institute
The organic solvent stated is preferably methanol or acetonitrile, and the proportion by weight of the sodium chloride and the herbal tea sample is preferably 0.5~
1:1, more preferably, the proportion by weight of the sodium chloride and the herbal tea sample is 1:1.
The organic solvent and the herbal tea sample are preferably 2~4mL with magnitude relation：1g, it is ensured that recovery rate and
Detection limit；The proportion by weight of the sodium chloride and the herbal tea sample is more preferably 1:1, wherein sodium chloride plays salting out,
The recovery rate of determinand can be improved.
Step (1) mesoscale eddies extraction rotating speed of the present invention is 1000~3000r/min, and vortex extraction time is no less than 2min,
As one kind of the invention preferred embodiment, the extraction of its mesoscale eddies is preferred to use commercially availableMS3 digital are vortexed
Blender B-mode.
Rotating speed is preferably 6000~9000 revs/min when being centrifuged in step (1) of the present invention, more preferably 8000 revs/min, from
The heart time is preferably 3~5min, more preferably 5min；As one kind of the invention preferred embodiment, it is preferred to use during centrifugation
Commercially available model SIGMA 3-18K centrifuge.
The parts by volume proportioning of supernatant described in step (1) of the present invention and the solution is preferably 1:1~2, more preferably 1:
1, wherein the concentration of the ammonium acetate solution is preferably 5~10mmol/L, more preferably 10mmol/L, pH value is 4.5~5.5, more
Good is 4.5.
As one kind of the invention preferred embodiment, pipette, extract supernatant can be used in step (1) of the present invention
In nitrogen blowpipe, ammonium acetate solution is then added, vortex mixed is uniform, and prepare liquid is made.
Such as step of the present invention (1) is settled to 50mL with methanol (or acetonitrile), takes 1.0mL to add 1.0mL in nitrogen blowpipe
10mmol/L ammonium acetate solutions (pH=4.5), it is 2.0mL to make final volume, and vortex mixed is uniform, and prepare liquid is made.
Solid herbal tea sample described in step (1) of the present invention and the organic solvent are preferably 1g with magnitude relation：25~
50mL, more preferably 1g：50mL；Described organic solvent is preferably methanol or acetonitrile, extracting section after constant volume described in step (1)
The parts by volume proportioning of liquid and the solution is preferably 1:1~2, more preferably 1：1, wherein the concentration of the ammonium acetate solution is preferably
5~10mmol/L, more preferably 10mmol/L, pH value are preferably 4.5~5.5, and more preferably 4.5.
The aperture of organic phase filter membrane is preferably 0.22 μm or 0.45 μm described in step (2) of the present invention, preferably commercially available
Organic filter membrane of nylon66 fiber material；Reverse-phase chromatographic column described in step (2) is preferably C18 chromatographic columns.
Wherein the aperture of organic phase filter membrane is 0.22 μm or 0.45 μm, can filter larger particles impurity, purifies sample.
Reverse-phase chromatographic column is C18 chromatographic columns.
Preferably using acetate-methanol solution as organic phase flow in step (2) of the present invention, the volume basis of wherein acetic acid contains
Amount is preferably 0.1%, and preferably using ammonium acetate solution as aqueous phase mobile phase, the pH of wherein solution is preferably 4.5~5.5, more preferably
For 4.5, concentration is preferably 5~10mmol/L, more preferably 10mmol/L.
The volumn concentration of acetic acid is 0.1% wherein in acetate-methanol solution, and the pH of ammonium acetate solution is 4.5, concentration
For 10mmol/L, under this flow visualizing, five kinds of main component chromatographic peak peak shapes of pappy shell are symmetrical, with more preferable separation
Degree and sensitivity.
Using five kinds of main components in ultra performance liquid chromatography-tandem mass spectrometer detection pappy shell in step (2) of the present invention
When morphine, codeine, thebaine, papaverine and coscopin, it is preferred to use polyion (is only to use standard to qualitative, external standard method
Material is quantified, without using internal standard substance.) quantitative, wherein morphine, papaverine are preferred to use 1 parent ion, and 3 daughter ions can be treated
Cause, thebaine, coscopin are using 1 parent ion, and the qualitative identification method of 2 daughter ions, morphine parent ion is m/z 286.3,
Daughter ion is m/z 165.0,181.0 and 201.1；Papaverine parent ion be m/z 340.3, daughter ion be m/z 171.1,
202.1 and 324.2；Codeine parent ion is m/z 300.2, and daughter ion is m/z 165.1,215.1；Thebaine parent ion is m/
Z 312.3, daughter ion is m/z 57.9,251.1；Coscopin parent ion be m/z 414.4, daughter ion be m/z 220.1,
353.2。
Step (2) of the present invention if in prepare liquid determinand content be above standard the range of linearity of curve, can with 5~
10mmol/L ammonium acetate solutions (pH=4.5~5.5) are with methanol according to volume ratio 1:1 mixed solution prepared carries out suitably dilute
Release.
With DB31/2010-2012《Papaverine in food product place of safety standard chafing dish food, morphine, coscopin, it can treat
The measure Liquid Chromatography-Tandem Mass Spectrometry of cause and thebaine》Compare, the invention has the characteristics that：
(1) ultrasonic extraction is heated：Liquid herbal tea, solid herbal tea sample, using heating ultrasonic extraction, make determinand abundant
It is dissolved in extract solution；DB31/2010-2012 sample treatments do not carry out appropriate heating, it is difficult to ensure that pappy shell is main
Composition is extracted by hydrochloric acid solution completely, it is possible that measurement not enough precisely, Lower result, poor repeatability situations such as；
(2) saltoutd under fluid sample weak basic condition extraction：Liquid herbal tea, is conducive in weakly alkaline solution with ion shape
The determinand formation molecular state that formula is present, under the conditions of addition sodium chloride is saltoutd, improves extraction of the methanol (or acetonitrile) to determinand
Rate is taken, after DB31/2010-2012 standards are scattered to samples with water such as sauce, soup stock, flavored oils, not adjusted pH value is just direct
Extracted with acetonitrile, poppy capsule alkaloid composition recovery rate may be made relatively low and its pre-treatment in the anhydrous magnesium sulfate that adds and
Anhydrous sodium acetate mixed-powder has certain suction-operated, Lower result to determinand；
(3) the inventive method is reliable, simple and quick, economical：Chafing dish food contains the materials such as grease, spices, and matrix is complicated,
Interference is big, it is necessary to further remove oil purification using PSA powder, C18 powder, PSA powder, C18 powder are expensive, therefore, testing cost compared with
Height, is purified with PSA powder, anhydrous magnesium sulfate, C18 powder, there is certain suction-operated to determinand, causes part determinand loss,
The rate of recovery is relatively low, and herbal tea sample substrate is relatively easy, by the advantage of mass spectrographic strong antijamming capability, and it is accurately fixed to carry out
Property confirmation, in pre-treatment step, adjust by pH, saltout, methanol (or acetonitrile) is extracted, can be removed largely after high speed centrifugation
Chaff interference, it is simple and quick, it is economical；
(4) the inventive method sensitivity is high：Herbal tea sample after processing more preferably using 1.0mL methanol (or acetonitrile) and
1.0mL 10mmol/L ammonium acetate solutions (pH=4.5) are mixed, and can improve chromatographic peak profile, improve the response of determinand, than
DB31/2010-2012 standards use acetonitrile solute effect more preferably；
(5) present invention is optimal (adjusts pH=with 0.1% acetate-methanol and 10mmol/L ammonium acetate solutions with glacial acetic acid
4.5) it is the pKa value according to morphine, codeine, thebaine, papaverine and coscopin as mobile phase, is flowed by adjusting aqueous phase
The pH of phase is more preferably 4.5, makes acquisition shorter analysis time, preferable separating degree and sensitivity.
Therefore, compared with prior art, the present invention has advantages below：
(1) the detection method of the invention opium poppy suitable for any liquid on the market, solid herbal tea sample
The detection of shell component, pre-treatment is simple, quick, and interference is few, and Qualitive test is accurately and reliably；
(2) ultrasonic extraction can make pappy shell active ingredient in herbal tea fully molten to liquid herbal tea sample in a heated condition
Solution；Solid herbal tea sample is extracted with methanol (or acetonitrile), is more beneficial for the dissolving of pappy shell main component, extraction effect is more preferably；
(3) pH value of regulation liquid herbal tea sample can make the composition to be measured in herbal tea in pappy shell by ion for alkalescent
State is converted to molecular state, and the extraction of methanol (or acetonitrile) is more beneficial under the conditions of sodium chloride is saltoutd, and improves recovery rate, Er Qie
There is higher separating degree and detection sensitivity in ultra performance liquid chromatography-tandem mass spectrum detection；
(3) when the flowing phase pH value that uses of the present invention is more preferably 4.5, the pH value of mobile phase is close to the pKa value of determinand, energy
Improve peak shape, improve separating degree and sensitivity；
(4) present invention is established simple, fast on the premise of illegal additive examination criteria reference in not yet having herbal tea
Ultra performance liquid chromatography-tandem mass spectrum quick detection side of illegal addition opium poppy shell component in speed, the accurate herbal tea of qualitative, quantitative
Method.
Brief description of the drawings
Fig. 1 is the mark-on sample total ion chromatogram of embodiment 1 (TIC figures)；
Fig. 2 is the morphine standard curve (5~800ng/mL) in embodiment 1-2；
Fig. 3 is the thebaine standard curve (1~200ng/mL) in embodiment 1-2；
Fig. 4 is the papaverine standard curve (1~200ng/mL) in embodiment 1-2；
Fig. 5 is the codeine standard curve (5~1000ng/mL) in embodiment 1-2；
Fig. 6 is the coscopin standard curve (1~200ng/mL) in embodiment 1-2；
Fig. 7 is the morphine multiple-reaction monitoring chromatogram (MRM chromatograms) in embodiment 1；
Fig. 8 is the codeine multiple-reaction monitoring chromatogram (MRM chromatograms) in embodiment 1；
Fig. 9 is the thebaine multiple-reaction monitoring chromatogram (MRM chromatograms) in embodiment 1；
Figure 10 is the papaverine multiple-reaction monitoring chromatogram (MRM chromatograms) in embodiment 1；
Figure 11 is the coscopin multiple-reaction monitoring chromatogram (MRM chromatograms) in embodiment 1；
Figure 12 is the positive total ion chromatogram of embodiment 2 (TIC figure)；
Figure 13 is the morphine multiple-reaction monitoring chromatogram (MRM chromatograms) in embodiment 2；
Figure 14 is the codeine multiple-reaction monitoring chromatogram (MRM chromatograms) in embodiment 2；
Figure 15 is the thebaine multiple-reaction monitoring chromatogram (MRM chromatograms) in embodiment 2；
Figure 16 is the papaverine multiple-reaction monitoring chromatogram (MRM chromatograms) in embodiment 2；
Figure 17 is the coscopin multiple-reaction monitoring chromatogram (MRM chromatograms) in embodiment 2.
Embodiment
The present invention is further illustrated below in conjunction with specific embodiment, but the scope of protection of present invention not office
It is limited to following embodiments.
Embodiment 1
The method of illegal addition pappy shell, comprises the following steps in the quick discriminating herbal tea that the present embodiment is provided：
(1) herbal tea sample about 50mL in bulk on the market is taken to be placed in 250mL beakers, at 40 DEG C with 40khz frequencies
Ultrasonic extraction 20 minutes, obtains homogeneous sample；
(2) take out and be cooled to room temperature, precision weighs uniform liquid herbal tea sample 5g (being accurate to 0.01g), is placed in 50mL
In plastic centrifuge tube, ammonia spirit or sodium hydroxide solution 5mL that pH is 8~9 are added, analytically pure methanol is added
10.0mL, adds sodium chloride 5g,MS3 digital turbine mixers were with 2000r/min vortex mixeds 3 minutes, with 8000
Rev/min centrifugation 5 minutes, layering accurately draws 1.0mL supernatant liquors with pipette in nitrogen blowpipe, then accurate to add
1.0mL 10mmol/L ammonium acetate solutions (pH=4.5), it is 2.0mL to make final volume, and vortex mixed is uniform, and prepare liquid is made；
(3) 1mL prepare liquids are drawn with 1mL disposable syringes, is filtered by 0.45 μm of aperture organic phase filter membrane in sample introduction
In bottle, close the lid, it is to be measured；If determinand content is above standard the range of linearity of curve in prepare liquid, 10mmol/L second can be used
Acid ammonium solution (pH=4.5) is with methanol according to volume ratio 1:1 mixed solution prepared carries out appropriate dilution；
(4) it is with 0.1% (volumn concentration) acetate-methanol solution and 10mmol/L ammonium acetate solutions (pH=4.5)
Mobile phase, the 2.0mm*50mm chromatogram post separations of Shim-pack XR-ODS III, using ultra performance liquid chromatography-tandem mass spectrometer
Five kinds of main component morphines, codeine, thebaine, opium poppies in pappy shell are detected under cation multiple-reaction monitoring (MRM) pattern
Alkali and coscopin；
Standard liquid is prepared and specific instrument condition is as follows：
1. the preparation of standard liquid
Using the liquid standard product of o2si companies of U.S. production, concentration is respectively：Morphine (CAS 57-27-2) concentration is
50mg/L, papaverine (CAS 61-25-6) concentration be 10mg/L, coscopin (CAS 912-60-7) concentration be 10mg/L, can
Treat because (CAS 76-57-3) concentration is 50mg/L and thebaine (CAS 115-37-7) concentration is 10mg/L.Morphine, it can treat
Cause, thebaine, papaverine and coscopin take 100 μ L to be placed in sample injection bottle respectively, with 10mmol/L ammonium acetate solutions (pH=4.5)
With methanol according to 1:The mixed solution that 1 volume ratio is made into is settled to 1mL, obtain mixed standard solution concentration be respectively 5mg/L,
5mg/L, 1mg/L, 1mg/L, 1mg/L, with 10mmol/L ammonium acetate solutions (pH=4.5) and methanol according to 1:1 volume ratio is made into
Mixed solution as solvent, morphine, papaverine are prepared into mixed standard solution dilution according to actual needs, coscopin, can be treated
Cause and thebaine standard curve, concentration range be respectively 5~800ng/mL, 1~200ng/mL, 1~200ng/mL, 5~
1000ng/mL, 1~200ng/mL, are detected using quantified by external standard method.Standard curve is as shown in figures 2-6.
2. liquid chromatogram and Mass Spectrometry Conditions
With 0.1% acetate-methanol solution (A phases) with 10mmol/L ammonium acetate solutions (B phases, pH=4.5) as mobile phase,
Flow velocity 0.2mL/min, gradient elution, the 2.0mm*50mm chromatogram post separations of Shim-pack XR-ODS III, 40 DEG C of column temperature, sample introduction
10 μ L are measured, with ultra performance liquid chromatography-tandem mass spectrometer cation multiple-reaction monitoring (MRM) mode detection.Gradient elution program
For：0~2.0min, 5%A phase；2.1~8.0min, 50%~80%A phase；8.1~11.0min, 5%A phase.
The mass spectrometric Qualitive test parameters of Waters UPLC-TQD：Morphine, codeine, thebaine, papaverine and coscopin
Five kinds of compositions, quantified by external standard method, morphine, papaverine are using 1 parent ion, 3 daughter ions, and codeine, thebaine, coscopin are adopted
With 1 parent ion, the qualitative identification method of 2 daughter ions, wherein morphine parent ion are m/z 286.3, and daughter ion is m/z
165.0th, 181.0 and 201.1；Papaverine parent ion is m/z 340.3, and daughter ion is m/z 171.1,202.1 and 324.2；Can
Treat that daughter ion is m/z 165.1,215.1 because parent ion is m/z 300.2；Thebaine parent ion is m/z 312.3, daughter ion
For m/z 57.9,251.1；Coscopin parent ion is m/z 414.4, and daughter ion is m/z 220.1,353.2.Morphine, papaverine,
The quota ion of coscopin, codeine and thebaine is respectively m/z 201.1,202.1,220.1,215.1,57.9, its minor
Ion is as qualitative ion.Multiple-reaction monitoring chromatogram (MRM chromatograms) is as illustrated in figures 7-11.
3. method detection limit
Using the liquid herbal tea sample addition morphine, papaverine, coscopin, codeine and thebaine standard of various effects
Product, addition concentration is respectively 50 μ g/kg, 10 μ g/kg, 10 μ g/kg, 50 μ g/kg, 10 μ g/kg.Grasped according to embodiment
Make, obtain chromatogram, signal to noise ratio is calculated to qualitative, quota ion chromatographic peak, signal to noise ratio (S/N) is all higher than 10.Therefore,
The detection limit of coffee, papaverine, coscopin, codeine and thebaine in herbal tea sample is respectively 50 μ g/kg, 10 μ g/kg, 10 μ
g/kg、50μg/kg、10μg/kg。
4. recovery of standard addition and precision
Selection is free of a kind of liquid herbal tea sample of determinand, adds three level standard materials respectively, papaverine, coscopin,
Thebaine pitch-based sphere is 50,100,250 μ g/kg, and morphine, codeine pitch-based sphere are 150,250,500 μ g/kg, and liquid is cool
The rate of recovery of tea is 73.8%~84.3%.The specific rate of recovery, precision are shown in Table 1.Mark-on sample total ion chromatogram (TIC
Figure) as shown in Figure 1.
The rate of recovery and precision of the liquid herbal tea of table 1
5. Qualitive test
Qualitive test：Sample and standard liquid are determined according to the condition of above-mentioned embodiment, if sample is all
Qualitative, quota ion occurs, and quota ion response peak is consistent with standard liquid；Simultaneously in sample qualitative ion pair abundance with
The relative abundance of standard liquid is than unanimously, relative abundance is provided than deviation no more than table 1, then can determine whether sample under its close concentration
In there is corresponding measured object.
When detecting morphine, codeine, thebaine, papaverine and coscopin simultaneously in herbal tea sample, you can judge herbal tea
In illegally with the addition of pappy shell.
With respect to the maximum allowable offset of abundance of ions during 2 qualitative determination of table
| Relative ion abundance | ＞ 50% | ＞ 20%~50% | ＞ 10%~20% | ﹤ 10% |
| The relative deviation of permission | ± 20% | ± 25% | ± 30% | ± 50% |
6. quantitative calculate
Calculation formula is：
X=C × V × f/m
Wherein：
X：Papaverine in sample, morphine, coscopin, the content (μ g/kg) of codeine and thebaine
C：The concentration (ng/mL) of tested component is obtained from standard working curve
m：Sampling quality (g)
V：Constant volume (mL)
f：Extension rate
As a result 1 is retained to decimal point.
7. actual sample is detected
Herbal tea sample in bulk totally 100 batch spot-check in the market, wherein the liquid herbal tea sample inspection of 2 batch cough-relieving effects
Go out five kinds of main components of pappy shell, wherein 1 batch morphine, papaverine, coscopin, codeine and thebaine content are respectively
314.7 μ g/kg, 92.8 μ g/kg, 154.7 μ g/kg, 55.9 μ g/kg, 150.9 μ g/kg；Another 1 batch morphine, papaverine, that can
Fourth, codeine and thebaine content are respectively 114.7 μ g/kg, 43.3 μ g/kg, 74.7 μ g/kg, are not detected, 80.7 μ g/kg.
Embodiment 2
Pappy shell have astringe the lung, puckery intestines, analgesic, cough-relieving the effects such as.Cough-relieving of some bad businessmans in order to improve herbal tea is imitated
Really, pappy shell is illegally added, its addition manner there may be directly addition pappy shell powder or the liquid of acquisition is boiled with pappy shell
Body is blent with herbal tea mix again.Detection for opium poppy shell component in solid kind herbal tea uses embodiment described below：
(1) solid herbal tea sample, weighs 1g (being accurate to 0.01g) solid sample and is placed in 50mL volumetric flasks, adds methanol
40mL covers bottle stopper, is placed in supersonic wave cleaning machine, and regulation temperature, with 40khz fixed frequencies ultrasound 25min, makes opium poppy for 50 DEG C
Shell main component is fully extracted, and is cooled to room temperature, with methanol constant volume to 50mL, takes 1.0mL to add 1.0mL in nitrogen blowpipe
10mmol/L ammonium acetate solutions (pH=4.5), it is 2.0mL to make final volume, and vortex mixed is uniform, and prepare liquid is made；
(2) 1mL prepare liquids are drawn with 1mL disposable syringes, is filtered by 0.45 μm of aperture organic phase filter membrane in sample introduction
In bottle, close the lid, it is to be measured；If determinand content is above standard the range of linearity of curve in prepare liquid, 10mmol/L second can be used
Acid ammonium solution (pH=4.5) is with methanol (according to volume ratio 1:1 mixed solution prepared carries out appropriate dilution.
(3) with 0.1% acetate-methanol solution and 10mmol/L ammonium acetate solutions (pH=4.5) for mobile phase, Shim-
The 2.0mm*50mm chromatogram post separations of pack XR-ODS III, are reacted the cation more using ultra performance liquid chromatography-tandem mass spectrometer
Five kinds of main component morphines, codeine, thebaine, papaverine and coscopin in detection pappy shell under (MRM) pattern of monitoring.If treating
Survey determinand content in liquid to be above standard the range of linearity of curve, 10mmol/L ammonium acetate solutions (pH=4.5) and methanol can be used
According to volume ratio 1:1 mixed solution prepared carries out appropriate dilution.
Standard liquid is prepared and specific instrument condition is as follows：
1. the preparation of standard liquid：It is same as Example 1.
2. liquid chromatogram and Mass Spectrometry Conditions：It is same as Example 1.
3. method detection limit：It is same as Example 1.
4. recovery of standard addition and precision
A kind of solid herbal tea sample of the selection without determinand, adds three level standard materials respectively, papaverine, that can
Fourth, thebaine pitch-based sphere are 50,100,250 μ g/kg, and morphine, codeine pitch-based sphere are 150,250,500 μ g/kg, solid
The herbal tea rate of recovery is 71.1%~83.6%.The specific rate of recovery, precision are shown in Table 3.
The rate of recovery and precision of the solid herbal tea of table 3
5. Qualitive test：It is same as Example 1.
6. quantitative calculate：It is same as Example 1.
7. actual sample is detected
Solid herbal tea sample is operated according to the method for embodiment 2, detection morphine, papaverine, coscopin, codeine and thebaine
Content is respectively 1314.7 μ g/kg, 253.8 μ g/kg, 226.7 μ g/kg, is not detected, 290.9 μ g/kg.Figure 12 is the sun of embodiment 2
Property sample total ion chromatogram (TIC chromatograms), Figure 13-17 be respectively the positive of embodiment 2 in morphine, codeine, the base of a fruit
Ba Yin, papaverine, the multiple-reaction monitoring chromatogram (MRM chromatograms) of coscopin.
Embodiment 3
Influence of the positive herbal tea sample without heating shaking test to result.
(1) liquid herbal tea sample 5g (being accurate to 0.01g) is weighed, is placed in 50mL plastic centrifuge tubes, it is 8~9 to add pH
Ammonia spirit or sodium hydroxide solution 5mL, add analytically pure methanol (or acetonitrile) 10.0mL, add sodium chloride 5g, whirlpool
Revolve incorporation time 3 minutes, centrifuged 5 minutes with 8000 revs/min, layering accurately draws 1.0mL supernatant liquors with pipette in nitrogen
Then accurate to add 1.0mL10mmol/L ammonium acetate solutions (pH=4.5) in blowpipe, it is 2.0mL to make final volume, is vortexed mixed
Close uniform, prepare liquid is made；Remaining step is operated by embodiment 1.
In positive 1, morphine, papaverine, coscopin, codeine and thebaine content are respectively 264.2 μ g/kg,
87.5 μ g/kg, 144.1 μ g/kg, are not detected, 139.91 μ g/kg；In positive 2, morphine, papaverine, coscopin, codeine
It is respectively 95.7 μ g/kg, 31.9 μ g/kg, 68.7 μ g/kg with thebaine content, does not detect, 75.8 μ g/kg.
The result of embodiment 3 shows, the result obtained without heating shaking test is than Lower result that embodiment 1 is obtained.
Embodiment 4
Sample is compareed by heating vibration, but not adjusting pH test result with embodiment 1.
(1) herbal tea sample about 50mL in bulk on the market is taken to be placed in 250mL beakers, at 40-50 DEG C with 40khz frequencies
Rate ultrasonic extraction 20 minutes, obtains homogeneous sample；
(2) take out and be cooled to room temperature, precision weighs uniform liquid herbal tea sample 5g (being accurate to 0.01g), is placed in 50mL
In plastic centrifuge tube, ultra-pure water 5mL is added, analytically pure methanol (or acetonitrile) 10.0mL is added, sodium chloride 5g is added, is vortexed
Incorporation time 3 minutes, is centrifuged 5 minutes with 8000 revs/min, and layering is accurately drawn 1.0mL supernatant liquors with pipette and blown in nitrogen
Guan Zhong, then accurate to add 1.0mL10mmol/L ammonium acetate solutions (pH=4.5), it is 2.0mL, vortex mixed to make final volume
Uniformly, prepare liquid is made；Remaining step is operated by embodiment 1.
Positive 1, morphine, papaverine, coscopin, codeine and thebaine content are respectively 214.6 μ g/kg, 87.8 μ
G/kg, 134.7 μ g/kg, is not detected, 141.7 μ g/kg；Positive 2, morphine, papaverine, coscopin, codeine and thebaine
Content is respectively 92.7 μ g/kg, 40.3 μ g/kg, 64.1 μ g/kg, is not detected, 72.4 μ g/kg.
Embodiment 4 does not adjust pH test result than the Lower result of embodiment 1.It is probably because part exists with ionic state
Determinand be not fully extracted.
A part of specific embodiment is enumerated above, and the present invention will be described, it is necessary to it is pointed out here that be specific real up and down
Apply example and be served only for that the present invention is further illustrated, do not represent limiting the scope of the invention.Other people are according to this hair
The bright nonessential modification of some made and adjustment still fall within protection scope of the present invention.Such as organic solvent can also be second
Nitrile, the ability such as use magnitude relation, centrifugal rotational speed, pH value, organic phase filter membrane aperture between ultrasonically treated temperature, time, each material
Field technique personnel can carry out appropriate adjustment in the range of application claims.
Claims (10)
1. a kind of method of illegal addition pappy shell in quick discriminating herbal tea, it is characterized in that comprising the following steps：
(1) liquid herbal tea sample is chosen, progress is ultrasonically treated, pappy shell main component in herbal tea sample is fully dissolved, and cools down
To room temperature, herbal tea sample is obtained, herbal tea sample is taken, it is adjusted in alkalescent, organic solvent is added and sodium chloride carries out vortex extraction
Take, then centrifuge again, be layered, take supernatant, ammonium acetate solution is then added in supernatant, mix, prepare liquid is made；
Or solid herbal tea sample is chosen, organic solvent is added, progress is ultrasonically treated, makes pappy shell main component quilt in herbal tea sample
Fully extract, obtain extract solution, extract solution is cooled to room temperature, constant volume takes part extract solution after constant volume to add ammonium acetate solution, mixed
It is even, prepare liquid is made；
(2) after prepare liquid is filtered through organic phase filter membrane, using acetate-methanol solution and ammonium acetate solution as mobile phase, through anti-
After phase chromatogram post separation, eluent is detected into five kinds of main components in pappy shell using ultra performance liquid chromatography-tandem mass spectrometer
Morphine, codeine, thebaine, papaverine and coscopin.
2. the method for illegal addition pappy shell in quick discriminating herbal tea according to claim 1, it is characterized in that：Step (1)
In it is ultrasonically treated when, regulation temperature be 40~50 DEG C with 15~25min of 40khz frequency ultrasounds.
3. the method for illegal addition pappy shell in quick discriminating herbal tea according to claim 1, it is characterized in that：Step (1)
In take herbal tea sample, add pH value and adjust it in alkalescent, wherein herbal tea sample for 8~9 ammonia spirit or sodium hydroxide solution
Ammonia spirit that product and pH value are 8~9 or sodium hydroxide solution are 1g with magnitude relation：1~2mL.
4. the method for illegal addition pappy shell in quick discriminating herbal tea according to claim 1, it is characterized in that：Step (1)
Described in the use magnitude relation of organic solvent and the herbal tea sample be 2~4mL：1g, wherein described organic solvent be methanol or
The proportion by weight of acetonitrile, the sodium chloride and the herbal tea sample is 0.5~1:1.
5. the method for illegal addition pappy shell in quick discriminating herbal tea according to claim 1, it is characterized in that：Step (1)
The rotating speed of mesoscale eddies extraction is 1000~3000r/min, and vortex extraction time is no less than 2min；During centrifugation rotating speed be 6000~
9000 revs/min, centrifugation time is 3~5min.
6. the method for illegal addition pappy shell in quick discriminating herbal tea according to claim 1, it is characterized in that：Step (1)
Described in the parts by volume proportioning of supernatant and the ammonium acetate solution be 1:1~2, wherein the concentration of the ammonium acetate solution is 5
~10mmol/L, pH value is 4.5~5.5.
7. the method for illegal addition pappy shell in quick discriminating herbal tea according to claim 1, it is characterized in that：Step (1)
Described in solid herbal tea sample and the organic solvent be 1g with magnitude relation：25~50mL, described organic solvent is methanol
Or acetonitrile, the parts by volume proportioning of part extract solution and the ammonium acetate solution is 1 after constant volume described in step (1):1~2, wherein
The concentration of the ammonium acetate solution is 5~10mmol/L, and pH value is 4.5~5.5.
8. the method for illegal addition pappy shell in quick discriminating herbal tea according to claim 1, it is characterized in that：Step (2)
Described in organic phase filter membrane aperture be 0.22 μm or 0.45 μm；Reverse-phase chromatographic column described in step (2) is C18 chromatographic columns.
9. the method for illegal addition pappy shell in quick discriminating herbal tea according to claim 1, it is characterized in that：Step (2)
In using acetate-methanol solution as organic phase flow, wherein the volumn concentration of acetic acid be 0.1%, made with ammonium acetate solution
For aqueous phase mobile phase, the wherein pH of ammonium acetate solution is 4.5~5.5, and concentration is 5~10mmol/L.
10. the method for illegal addition pappy shell in quick discriminating herbal tea according to claim 1, it is characterized in that：Step (2)
Five kinds of main component morphines, codeine, thebaine, small-mouthed jars in middle use ultra performance liquid chromatography-tandem mass spectrometer detection pappy shell
When pavine and coscopin, using polyion to qualitative, quantified by external standard method, wherein morphine, papaverine are using 1 parent ion, 3 sons
Ion, codeine, thebaine, coscopin are using 1 parent ion, and the qualitative identification method of 2 daughter ions, morphine parent ion is m/
Z 286.3, daughter ion is m/z 165.0,181.0 and 201.1；Papaverine parent ion is m/z 340.3, and daughter ion is m/z
171.1st, 202.1 and 324.2；Codeine parent ion is m/z 300.2, and daughter ion is m/z 165.1,215.1；Thebaine it is female from
Son is m/z 312.3, and daughter ion is m/z 57.9,251.1；Coscopin parent ion is m/z 414.4, and daughter ion is m/z
220.1、353.2。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710259454.4A CN107102075B (en) | 2017-04-20 | 2017-04-20 | Method for rapidly identifying illegally added poppy shells in herbal tea |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710259454.4A CN107102075B (en) | 2017-04-20 | 2017-04-20 | Method for rapidly identifying illegally added poppy shells in herbal tea |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107102075A true CN107102075A (en) | 2017-08-29 |
| CN107102075B CN107102075B (en) | 2020-02-04 |
Family
ID=59658140
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710259454.4A Active CN107102075B (en) | 2017-04-20 | 2017-04-20 | Method for rapidly identifying illegally added poppy shells in herbal tea |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107102075B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108717089A (en) * | 2018-05-25 | 2018-10-30 | 三明出入境检验检疫局综合技术服务中心 | A kind of rapid detection method of papaverine, coscopin and thebaine |
| CN108956798A (en) * | 2018-04-27 | 2018-12-07 | 青海省药品检验检测院 | A kind of morphine hydrochloride preparation content determining method |
| CN111208231A (en) * | 2020-01-20 | 2020-05-29 | 广东省药品检验所（广东省药品质量研究所、广东省口岸药品检验所） | Method for simultaneously detecting six illegally added compounds in plant beverage |
| CN113075326A (en) * | 2021-03-31 | 2021-07-06 | 周迎春 | Method for rapidly determining poppy shell alkaloid by liquid chromatography-mass spectrometry-internal standard method |
| CN114354781A (en) * | 2021-12-15 | 2022-04-15 | 广西桂林锐德检测认证技术有限公司 | Method for simultaneously determining 5 illegal additives in food |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100051801A1 (en) * | 2008-09-03 | 2010-03-04 | Erfurth Stephen C | Methods and systems for analyzing medication levels in a sample |
| CN104897768A (en) * | 2015-06-19 | 2015-09-09 | 南京工业大学 | Method for rapidly determining morphine, codeine, thebaine, papaverine and narcotine in hot-pot food |
| CN105588904A (en) * | 2015-06-19 | 2016-05-18 | 英格尔检测技术服务（上海）有限公司 | Determination method for papaverine in hot pot food |
-
2017
- 2017-04-20 CN CN201710259454.4A patent/CN107102075B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100051801A1 (en) * | 2008-09-03 | 2010-03-04 | Erfurth Stephen C | Methods and systems for analyzing medication levels in a sample |
| CN104897768A (en) * | 2015-06-19 | 2015-09-09 | 南京工业大学 | Method for rapidly determining morphine, codeine, thebaine, papaverine and narcotine in hot-pot food |
| CN105588904A (en) * | 2015-06-19 | 2016-05-18 | 英格尔检测技术服务（上海）有限公司 | Determination method for papaverine in hot pot food |
Non-Patent Citations (4)
| Title |
|---|
| 刘敏敏等: "液相色谱-串联质谱法检测火锅底料中罂粟壳", 《食品研究与开发》 * |
| 林黛琴等: "高效液相色谱-串联质谱法快速测定食品中５种罂粟壳生物碱", 《质谱学报》 * |
| 王力清等: "超高效液相色谱-串联质谱法高通量快速测定调料中罂粟壳生物碱含量", 《食品与发酵工业》 * |
| 皮立等: "高效液相色谱-荧光检测法测定罂粟籽和火锅汤料中的罂粟碱", 《色谱》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108956798A (en) * | 2018-04-27 | 2018-12-07 | 青海省药品检验检测院 | A kind of morphine hydrochloride preparation content determining method |
| CN108717089A (en) * | 2018-05-25 | 2018-10-30 | 三明出入境检验检疫局综合技术服务中心 | A kind of rapid detection method of papaverine, coscopin and thebaine |
| CN111208231A (en) * | 2020-01-20 | 2020-05-29 | 广东省药品检验所（广东省药品质量研究所、广东省口岸药品检验所） | Method for simultaneously detecting six illegally added compounds in plant beverage |
| CN113075326A (en) * | 2021-03-31 | 2021-07-06 | 周迎春 | Method for rapidly determining poppy shell alkaloid by liquid chromatography-mass spectrometry-internal standard method |
| CN113075326B (en) * | 2021-03-31 | 2024-03-15 | 周迎春 | Method for rapidly determining poppy shell alkaloid by liquid chromatography-mass spectrometry (LIMS) -internal standard method |
| CN114354781A (en) * | 2021-12-15 | 2022-04-15 | 广西桂林锐德检测认证技术有限公司 | Method for simultaneously determining 5 illegal additives in food |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107102075B (en) | 2020-02-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107102075A (en) | A kind of method of illegal addition pappy shell in quick discriminating herbal tea | |
| Cai et al. | Deep eutectic solvents used as the green media for the efficient extraction of caffeine from Chinese dark tea | |
| CN108459096B (en) | Heishanyin tablet extract and ginseng radix aconiti lateralis preparata injection prepared by real-time release method in automatic extraction process of Heishanyin tablet | |
| CN103330758A (en) | Peony and liquorice soup formula granule, preparation method and detection method of peony and liquorice soup formula granule | |
| CN109839458B (en) | Method for detecting sodium picosulfate in food | |
| CN112858508B (en) | Method for determining furfural compounds in coffee product | |
| CN105548433B (en) | The method that liquid chromatography tandem mass spectrometry detects various non-protein nitrogen-containing compound contents in milk simultaneously | |
| CN106153810A (en) | A kind of preparation method and application of stem and leaf of Radix Ginseng total saponins reference extract | |
| CN104155383B (en) | The detection method of blue or green Pu granule | |
| CN105510312A (en) | Orlistat rapid detection method | |
| CN105548161B (en) | A kind of quick determination method of Rimonabant | |
| CN103728380A (en) | Method for ionic liquid extraction/HPLC detection of fleroxacin residues in aquatic product | |
| CN102759598A (en) | Quality detection method for 29-componnet stagnation dissipation powder as Tibetan medicinal composition and preparations thereof | |
| CN103105446B (en) | Method for enriching beta-lactam penicillins antibiotics in liquid milk | |
| CN111929378B (en) | Method for measuring content of 6 index components of gastrodia elata in Qingda granules | |
| CN104483411B (en) | A kind of capsule of weeping forsythia and the detection method containing capsule of weeping forsythia product | |
| CN115372497A (en) | Method for measuring content of 6 alkaloid components in small collateral activating pills | |
| CN106198414A (en) | The detection method of olive oil Determination of Chlorophyll copper sodium | |
| CN106198821A (en) | A kind of method detecting the residual of sulfa antibiotics in milk | |
| CN109387579A (en) | The method for measuring levamisol in dairy products | |
| CN113702522A (en) | Method for constructing characteristic spectrum of periostracum cicadae medicinal material, and detection method of traditional Chinese medicine decoction and traditional Chinese medicine formula granules of periostracum cicadae medicinal material | |
| CN102680606B (en) | Method for measuring sodium formaldehyde sulfoxylate in food | |
| CN106979987B (en) | The HPLC detection method of main active in Lanhuayao | |
| CN109655549A (en) | A kind of method of quick detection fish Malachite Green | |
| CN109884211A (en) | The measuring method of glycerol content in a kind of detection honey |
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 |