CN107966515A - The method that ultra performance liquid chromatography tandem mass spectrometry measures poppy capsule alkaloid and industrial dye at the same time - Google Patents
The method that ultra performance liquid chromatography tandem mass spectrometry measures poppy capsule alkaloid and industrial dye at the same time Download PDFInfo
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Abstract
The present invention provides a kind of method that ultra performance liquid chromatography tandem mass spectrometry measures poppy capsule alkaloid and industrial dye at the same time, sample is extracted using acetonitrile solution, acetonitrile layer rotary evaporation after anhydrous magnesium sulfate is dehydrated is concentrated to dryness, with acetonitrile, formic acid water dissolved residue, sample liquid crosses film, detected using ultra performance liquid chromatography tandem mass spectrometry, quantified by external standard method.5 kinds of poppy capsule alkaloids and 6 kinds of industrial dyes have good linear relationship in the concentration range of 2.5~100 μ g/L, and the average recovery rate of sample is 72.2%~108% under 3 pitch-based spheres, relative standard deviation 2.5~9.6%.In actual sample detection, detection Acid Orange II positive 3, content is 0.0159~74.2mg/kg.This method is accurate, sensitive, quick, is to detect the effective ways of poppy capsule alkaloid and industrial dye in snack products and chafing dish bottom flavorings at the same time.
Description
Technical field
The present invention relates to detection technique field, more particularly to a kind of ultra performance liquid chromatography tandem mass spectrometry to measure small-mouthed jar at the same time
The method of grain shell alkaloid and industrial dye.
Background technology
Snack products are the general names of a kind of food with notable local characteristic, are not exclusively used as breakfast, food,
It can be interspersing between dinner or feast, be one of cuisines indispensable in quite a few people life；And chafing dish then one
Directly sweep the country, extensively liked by masses.Just due to this, Partial Food processor, which deceives, cares for state's laws, regulation and people's body
Health, unscrupulous addition pappy shell and industrial dye so as to attract frequent customer, increase sales volume so that food " more good-looking, nicier ",
Play one's own game, also so that chafing dish bottom flavorings and snack products become the severely afflicated area of this illegal addition of two classes.
Pappy shell contains more than the 30 kinds of alkaloid such as morphine, codeine, coscopin, papaverine, and people once eat and with the addition of
The food of pappy shell, will detect the poisonous substances such as morphine in vivo, cause the physical examinations such as conscription can not pass, long-term consumption will be to nerve
System, digestive system cause damage, and the symptoms such as endocrine disorder occur, or even addicted.Industrial dye includes tonyred, rose
Red B (rhodamine B), Acid Orange II etc., are used primarily in the dyeing of the industrial goods such as fur, polyamide fibre, paper, mostly with carcinogenic
Or teratogenesis.So China was issued in 2009《The non-edible material from soybeans of possible illegal addition and the food easily abused in food
Additive list》, prohibite pappy shell and tonyred, Wang Jinhuang (main component is basic orange II), rose red b (i.e. Luo Dan
Bright B), alkaline bright yellow, Acid Orange II, the industrial dye such as basic yellow be applied to food processing, wherein pappy shell, which is labelled with, to add
The Main Foods kind that the Main Foods kind added has chafing dish bottom flavorings and the industrial dye such as snack class, tonyred and may add, which has, to be contained
The food of capsicum class, Beancurd sheet, flavouring, salt down halogen meat products etc..
To prevent and the illegal addition in food of effective monitoring poppy capsule alkaloid and industrial dye, it is necessary to establish quick
The method that poppy capsule alkaloid and industrial dye are detected while accurate, to ensure food safety and consumer health.Pappy shell
The detection of alkaloid mainly has thin-layered chromatography, oscilloscopic polarography, immunochromatographic method, liquid chromatography, gas chromatography, liquid phase
Combined gas chromatography mass spectrometry；The detection method of industrial dye mainly has liquid chromatography, gas chromatography-mass spectrography, liquid phase in food
Chromatography-tandem mass spectrometry.China is directed to poppy capsule alkaloid and industrial dye in food and has promulgated relevant examination criteria respectively,
But the inadequate system of these standards, the particularly detection of industrial dye is mostly single variety, and have that certain methods step is complex,
Lack and detect the method for poppy capsule alkaloid and industrial dye in food at the same time
It is considered as desirable by the inventor to extract object, the detection of ultra performance liquid chromatography tandem mass spectrometer, is established using acetonitrile solution
Measure the method for 5 kinds of poppy capsule alkaloids and 6 kinds of industrial dyes in snack products and chafing dish bottom flavorings at the same time.
The content of the invention
Present invention solves the technical problem that it is that provide a kind of ultra performance liquid chromatography tandem mass spectrometry measures snack at the same time
The method of 5 kinds of poppy capsule alkaloids and 6 kinds of industrial dyes in based food and chafing dish bottom flavorings, this method are accurate, sensitive, quick.
In view of this, the present invention provides a kind of ultra performance liquid chromatography tandem mass spectrometry to measure 5 kinds of small-mouthed jars in food at the same time
The method of grain shell alkaloid and 6 kinds of industrial dyes, comprises the following steps：Sample is placed in centrifuge tube by step a), add water and
Acetonitrile mixes, and then adds anhydrous sodium acetate, shakes up immediately, then the concussion 1-3min that is vortexed, and 3-8min is centrifuged with 10000r/min,
Temperature is 3-5 DEG C, takes supernatant to cross anhydrous magnesium sulfate, and filtrate rotates evaporation and concentration to dry, addition 1-3mL second at 35-45 DEG C
The mixed solution dissolved residue of nitrile and 0.1% formic acid water, solution are transferred completely into centrifuge tube, and 1- is centrifuged with 4000r/min
The volume ratio of 3min, excessively 0.22 μm of filter membrane, the acetonitrile and formic acid water is 95:5；Step b) is connected using ultra performance liquid chromatography
Mass spectrography analyzes the sample after step a processing, while detects the kind of the poppy capsule alkaloid in sample and industrial dye
Class and content.
Preferably, 5 kinds of poppy capsule alkaloids are papaverine, morphine, codeine, coscopin, thebaine.
Preferably, 6 kinds of industrial dyes are SudanⅣ, rhodamine B, Acid Orange II, basic orange II, alkali
Property Huang 1, basic flavine O.
Preferably, in step a, the volume ratio of water and acetonitrile is 1：5.
Preferably, in step a, the mass ratio of the sample, anhydrous sodium acetate and anhydrous magnesium sulfate is 2:1.5:
6。
Preferably, standard solution is prepared as follows：Appropriate standard substance is weighed respectively, with acetonitrile or second
Nitrile water dissolves and uses acetonitrile constant volume, is configured to single mark storing solution of 100mg/L, then be configured to 10mg/L hybrid standard it is molten
Liquid, is diluted step by step using preceding.
Preferably, in step b, liquid phase chromatogram condition is：Chromatographic column：SB-C18RRHD1.8 μm, 2.1mm ×
50mm；Column temperature：35℃；Sample size：2μL；Mobile phase：A is methanol, and B is 0.1% formic acid water, gradient elution program：0-4min,
5%A, flow velocity 0.2mL/min；4.5-7min, 60%-95%A, flow velocity 0.3mL/min-0.4mL/min；7.1-11min, 95%
A, flow velocity 0.5mL/min；11.5-15min 5%A, flow velocity 0.2mL/min.
Preferably, in step b, Mass Spectrometry Conditions are：Ion gun：Electric spray ion source, Acid Orange II using bear from
Subpattern, remaining uses positive ion mode, and negative ions gather at the same time；550 DEG C of ion source temperature, gas curtain gas 35psi, atomization
Gas 55psi, heating auxiliary gas 55psi, collision gas：Medium, ion gun spray voltage 5500V (- 4500V)；Scan mode：It is more
Reaction monitoring.
Preferably, in step b, 5 kinds of poppy capsule alkaloids and 6 kinds of industrial dye mass spectrometry parameters are as shown in table 1：
15 kinds of poppy capsule alkaloids of table and 6 kinds of industrial dye mass spectrometry parameters
Preferably, 5 kinds of poppy capsule alkaloids and 6 kinds of industrial dyes are in the concentration range of 2.5~100 μ g/L,
With good linear relationship.
The present invention is using ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) while measures snack products and fire
5 kinds of poppy capsule alkaloids and 6 kinds of industrial dyes in flavoring food.Sample is extracted using acetonitrile solution, and acetonitrile layer is through anhydrous magnesium sulfate
Rotary evaporation is concentrated to dryness after dehydration, and with acetonitrile, formic acid water dissolved residue, sample liquid is crossed film, connected using ultra performance liquid chromatography
Mass spectrography detects, quantified by external standard method.5 kinds of poppy capsule alkaloids and 6 kinds of industrial dyes in the concentration range of 2.5~100 μ g/L,
With good linear relationship (related coefficient is all higher than 0.9980), the average recovery rate of sample is under 3 pitch-based spheres
72.2%~108%, relative standard deviation 2.5~9.6%, method detection limit papaverine, coscopin, thebaine, rose red b,
Acid Orange II, basic flavine O, basic yellow 1 are 1.0 μ g/kg, and morphine, codeine, basic orange II, SudanⅣ are 2.5 μ g/kg.
In actual sample detection, detection Acid Orange II positive 3, content is 0.0159~74.2mg/kg.This method is accurate, spirit
It is quick, quick, it is to detect the effective ways of poppy capsule alkaloid and industrial dye in snack products and chafing dish bottom flavorings at the same time..
Brief description of the drawings
Fig. 1 is the multiple-reaction monitoring chromatogram for the morphine that 10 μ g/kg are added in blank chafing dish bottom flavorings；
Fig. 2 is the multiple-reaction monitoring chromatogram for the codeine that 10 μ g/kg are added in blank chafing dish bottom flavorings；
Fig. 3 is the multiple-reaction monitoring chromatogram for the thebaine that 10 μ g/kg are added in blank chafing dish bottom flavorings；
Fig. 4 is the multiple-reaction monitoring chromatogram for the papaverine that 10 μ g/kg are added in blank chafing dish bottom flavorings；
Fig. 5 is the multiple-reaction monitoring chromatogram for the Acid Orange II that 10 μ g/kg are added in blank chafing dish bottom flavorings；
Fig. 6 is the multiple-reaction monitoring chromatogram for the coscopin that 10 μ g/kg are added in blank chafing dish bottom flavorings；
Fig. 7 is the multiple-reaction monitoring chromatogram for the basic flavine O that 10 μ g/kg are added in blank chafing dish bottom flavorings；
Fig. 8 is the multiple-reaction monitoring chromatogram for the basic yellow 1 that 10 μ g/kg are added in blank chafing dish bottom flavorings；
Fig. 9 is the multiple-reaction monitoring chromatogram for the basic orange II that 10 μ g/kg are added in blank chafing dish bottom flavorings；
Figure 10 is the multiple-reaction monitoring chromatogram for the rhodamine B that 10 μ g/kg are added in blank chafing dish bottom flavorings；
Figure 11 is the multiple-reaction monitoring chromatogram for the SudanⅣ that 10 μ g/kg are added in blank chafing dish bottom flavorings.
Embodiment
For a further understanding of the present invention, the preferred embodiment of the invention is described with reference to embodiment, still
It should be appreciated that these descriptions are simply further explanation the features and advantages of the present invention, rather than to the claims in the present invention
Limitation.The embodiment of the invention discloses a kind of ultra performance liquid chromatography tandem mass spectrometry to measure 5 kinds of pappy shell lifes in food at the same time
The method of alkaloids and 6 kinds of industrial dyes, comprises the following steps：Sample is placed in centrifuge tube by step a), adds water and acetonitrile mixes
It is even, anhydrous sodium acetate is then added, is shaken up immediately, then the concussion 1-3min that is vortexed, 3-8min is centrifuged with 10000r/min, temperature is
3-5 DEG C, take supernatant to cross anhydrous magnesium sulfate, filtrate rotates evaporation and concentration to dry at 35-45 DEG C, add 1-3mL acetonitriles with
The mixed solution dissolved residue of 0.1% formic acid water, solution are transferred completely into centrifuge tube, and 1-3min, mistake are centrifuged with 4000r/min
The volume ratio of 0.22 μm of filter membrane, the acetonitrile and formic acid water is 95:5；Step b) utilizes ultra performance liquid chromatography tandem mass spectrometry
Sample after step a processing is analyzed, while detects the species of the poppy capsule alkaloid in sample and industrial dye and contains
Amount.
Preferably, 5 kinds of poppy capsule alkaloids are papaverine, morphine, codeine, coscopin, thebaine；Institute
6 kinds of industrial dyes are stated as SudanⅣ, rhodamine B, Acid Orange II, basic orange II, basic yellow 1, basic flavine O.
Preferably, in step a, the volume ratio of water and acetonitrile is 1：5, the sample, anhydrous sodium acetate and anhydrous
The mass ratio of magnesium sulfate is 2:1.5:6.
Preferably, standard solution is prepared as follows：Appropriate standard substance is weighed respectively, with acetonitrile or second
Nitrile water dissolves and uses acetonitrile constant volume, is configured to single mark storing solution of 100mg/L, then be configured to 10mg/L hybrid standard it is molten
Liquid, is diluted step by step using preceding.
Preferably, in step b, liquid phase chromatogram condition is：Chromatographic column：SB-C18RRHD1.8 μm, 2.1mm ×
50mm；Column temperature：35℃；Sample size：2μL；Mobile phase：A is methanol, and B is 0.1% formic acid water, gradient elution program：0-4min,
5%A, flow velocity 0.2mL/min；4.5-7min, 60%-95%A, flow velocity 0.3mL/min-0.4mL/min；7.1-11min, 95%
A, flow velocity 0.5mL/min；11.5-15min 5%A, flow velocity 0.2mL/min.
In step b, Mass Spectrometry Conditions are：Ion gun：Electric spray ion source, Acid Orange II use negative ion mode, remaining is equal
Using positive ion mode, negative ions gather at the same time；550 DEG C of ion source temperature, gas curtain gas 35psi, atomization gas 55psi, heating
Aid in gas 55psi, collision gas：Medium, ion gun spray voltage 5500V (- 4500V)；Scan mode：Multiple-reaction monitoring.
5 kinds of poppy capsule alkaloids and 6 kinds of industrial dyes have well linear in the concentration range of 2.5~100 μ g/L
Relation.
From above scheme as can be seen that the present invention is using acetonitrile solution extraction object, ultra performance liquid chromatography series connection matter
Spectrometer measures, and pre-treatment is not required to use the small column purifications of SPE, simple and direct quick, and high sensitivity, accuracy and precision are good, are same
When detect snack products and poppy capsule alkaloid and the effective ways of industrial dye in chafing dish bottom flavorings.
For a further understanding of the present invention, technical solution provided by the invention is carried out specifically with reference to embodiment
Bright, protection scope of the present invention is not limited by the following examples.
The raw material that the embodiment of the present invention uses is purchased in market.
Embodiment
1.1 instruments and reagent
(Aglient 1290-AB SCIEX QTRAP 5500, AB SCIEX are public for ultra performance liquid chromatography tandem mass spectrometer
Department)；High speed freezing centrifuge (SIGMA 3-18KS, SIGMA companies)；Vortex concussion instrument (MS 3B, IKA companies)；Rotary evaporation
Instrument (DPE-2120, EYELA company).
Papaverine, morphine, codeine, coscopin, thebaine, SudanⅣ, rhodamine B, Acid Orange II, basic orange II,
(purity is equal for basic yellow 1>95%, Dr.E company), basic flavine O (purity 81.0%, ACHEMTEK companies).
Methanol, acetonitrile, formic acid (chromatographically pure, CNW Technologies GmbH), (analysis is pure, and western Gansu Province science is public for acetonitrile
Department), anhydrous sodium acetate (analyzes pure, Chinese medicines group), and anhydrous magnesium sulfate (analyzes pure, CNW Technologies GmbH), and water is
Ultra-pure water.
1.2 experimental method
1.2.1 standard solution is prepared
Appropriate standard substance is weighed respectively, with acetonitrile or acetonitrile water (1：1) dissolving and (SudanⅣ is with third with acetonitrile constant volume
Ketone dissolves, with acetonitrile constant volume), be configured to single mark storing solution of 100mg/L, then be configured to 10mg/L hybrid standard it is molten
Liquid, is diluted step by step using preceding.
1.2.2 sample preparation
Representative sample 200g is taken, is crushed uniformly, loads clean container as sample, sealing, and indicate mark.
1.2.3 pre-treatment
Weigh uniform sample 2.00g to be placed in 50mL centrifuge tubes, add 3mL water, the mixing of 15mL acetonitriles, it is anhydrous to add 1.5g
Sodium acetate, shakes up immediately, then the concussion 2min that is vortexed, and centrifuges 5min with 10000r/min, temperature is 3-5 DEG C, takes supernatant to cross 6g
Anhydrous magnesium sulfate, filtrate rotate evaporation and concentration to dry ,+0.1% formic acid water (95 of addition 2.0mL acetonitriles at 40 DEG C:5,V:V it is) molten
Residue is solved, solution is transferred completely into 10mL centrifuge tubes, and 2min is centrifuged with 4000r/min, crosses 0.22 μm of filter membrane, to be determined.
1.2.4 Liquid Chromatography-Tandem Mass Spectrometry condition
(1) liquid phase chromatogram condition
Chromatographic column：1.8 μm of SB-C18RRHD, 2.1mm × 50mm (Agilent companies)；Column temperature：35℃；Sample size：2μ
L.Mobile phase：A is methanol, and B is 0.1% formic acid water, gradient elution program：0-4min, 5%A, flow velocity 0.2mL/min；4.5-
7min, 60%-95%A, flow velocity 0.3mL/min-0.4mL/min；7.1-11min, 95%A, flow velocity 0.5mL/min；11.5-
15min, 5%A, flow velocity 0.2mL/min.
(2) Mass Spectrometry Conditions
Ion gun：Electric spray ion source (ESI), Acid Orange II use negative ion mode, remaining uses positive ion mode,
Negative ions gather at the same time；550 DEG C of ion source temperature, gas curtain gas 35psi, atomization gas 55psi, heating auxiliary gas 55psi, collision
Gas：Medium, ion gun spray voltage 5500 (- 4500) V；Scan mode：Multiple-reaction monitoring (MRM), specific mass spectrometry parameters are shown in
Table 1.
15 kinds of poppy capsule alkaloids of table and 6 kinds of industrial dye mass spectrometry parameters
* quota ion (Quantitativeion)
As a result with analysis
2.1 chromatographic columns and Optimization of mobile phase
Test the different types of chromatographic column such as C8, C18, Hillic, under the conditions of identical mobile phase, the peak shape of Hillic columns
Overall poor, basic orange II, basic yellow 1, SudanⅣ seriously trail；And C8Basic yellow 1, the peak shape of SudanⅣ are symmetrical on column
Property is bad, and signal-to-noise ratio compares C18Column wants low more than 1/2；C18Using 1.8 μm of 2.1mm × 50mm of SB-C18RRHD be in column it is optimal,
Preferable to all objects reservation in addition to morphine, separating effect is also preferable, sensitivity highest, more reactions of each standard substance
Monitoring (MRM) chromatogram is shown in Fig. 1-Figure 11, and retention time is shown in Table 3.Mobile phase has investigated methanol：0.1% formic acid water and acetonitrile：
20mmol/L ammonium acetates (pH4.5)[7], peak shape and separating effect are very nearly the same under the conditions of finding two kinds, but the former sensitivity is more
Height, therefore select methanol：0.1% formic acid water is as mobile phase.Since SudanⅣ has stronger reservation on a cl 8 column, to allow it
Faster elute, flow velocity increases to 0.5mL/min and keeps 4min after the outflow of other compounds；When eluting SudanⅣ
Between section, compare 100% methanol and methanol：0.1% formic acid water (95：5, V:V) two ways, it is found that the sensitivity of the latter is higher by
Nearly 1 times, therefore ratio as retaining in SudanⅣ appearance time.
2.2 Pretreatments optimize
2.2.1 extracts reagent selects
If select the high extractant of water-based content such as 1% trichloroacetic acid (or 0.1moL/L hydrochloric acid)：Acetonitrile (8：2, V:
V), n-hexane degreasing need to be used, and using n-hexane degreasing SudanⅣ can be made to lose more than 80% through experiment, because tonyred
IV is more soluble in n-hexane；And select the high solution of ethane nitrile content (acetonitrile accounts for more than 80%), then solve grease substantially
Puzzlement problem, acetonitrile layer separates thoroughly with oil reservoir after centrifugation, and extraction effect is also preferable.
2.2.2 dissolved residue solution selects
The collocation of acetonitrile, methanol respectively with water (or 0.1% formic acid water) different proportion is employed as the molten of dissolved residue
Liquid, as a result with the formic acid water of acetonitrile+0.1% (95:5,V:V best results).
2.2.3 purification condition selects
C18, the PSA used in QuEChERS is considered first, and experiment finds C18, PSA energy adsorbing contaminant, but also adsorbs
Poppy capsule alkaloid and pigment, the particularly absorption to pigment are obvious, are made comparisons with Acid Orange II positive, same
Under experimental condition, the examination for respectively making three samples is purified and is not purified using 150mgC18,60mgPSA in last sample liquid
Test, the result is that the peak area average specific of purification group does not have what is purified to reduce more than 20%, it is clear that using C18, PSA as purification
The rate of recovery can be reduced.
Furthermore cleaning test is carried out using the SPE such as PCX, MCX, HLB pillars, but also all without the preferable effect of acquirement.
PCX pillars have most of pigment a preferable reservation, after the elution of 5mL5% ammonium hydroxide methanol, then with 3mL acetone and dichloromethane (1:
1) elute, remain to wash the papaverine of lower significant proportion, coscopin, thebaine, SudanⅣ, rhodamine B, basic orange II, alkalescence
Huang 1, but basic flavine O could not elute always, and its rate of recovery is almost 0；Using HLB pillars, rhodamine B, Basic Orange
II rate of recovery is less than 30%；It is that Acid Orange II is difficult to retain using MCX pillars, the rate of recovery is less than 25%, and SudanⅣ, sieve
Red bright B is combined with pillar to be tightly difficult to very much elute.
To sum up, above purifying step does not apply to this test method.
2.2.4 matrix effect is investigated
Respectively by the use of the sample liquid of blank sample and mobile phase as dilution, the standard working curve for preparing same concentrations carries out
Measure, has investigated the Ion response value with the compound of concentration different matrix, the matrix effect of analyzing liquid sample, the formula of use
For：ME=B/A, wherein A and B represent peak area of the object in mobile phase and vehicle solution respectively, if ME ＜ 1, say
Bright matrix can suppress the response of object；ME ＞ 1, then illustrate that response of the matrix to object has humidification；ME=1 is represented not
There are matrix effect[16], it is generally understood that ME values are that matrix effect is not present in 0.85-1.15.The 5 kinds of pappy shell biologies measured
The matrix effect of alkali and 6 kinds of industrial dyes in different matrix is shown in Table 2, and since morphine appearance is more early, substrate inhibition is more obvious,
But on the whole, matrix effect not especially severe, prepares standard working curve by using bare substrate, keeps standard liquid and sample liquid
With identical ionization conditions, matrix effect, accuracy, the stability of not impact analysis result can be substantially eliminated.Illustrate this
Though test method does not take the step of further purification, and feasible.
Matrix effect of the 5 kinds of poppy capsule alkaloids and 6 kinds of industrial dyes of 2 various concentrations of table in different blank sample matrix
Should
The range of linearity and detection limit of 2.3 methods
2.3.1 standard working curve and related coefficient
Mixing chafing dish bottom flavorings, chafing dish food, sweet sour pork lit-chi style, boil in water for a while, then dress with soy, vinegar, etc. the different blank sample extracting solution of 4 kinds of meat etc. as mark
The dilute solution of quasi- material, it is 1.0,2.5,5.0,10.0,25.0,50.0,100.0 μ g/L matrix to make series mass concentration
Standard working solution, is measured by test method, draws standard curve.Equation of linear regression and related coefficient are shown in Table 3.
2.3.2 detection limit
Add standard substance in blank sample to be measured, it is detection limit to choose the additive amount that signal-to-noise ratio is 3, side
Method detection limit papaverine, coscopin, thebaine, rhodamine B, Acid Orange II, basic flavine O, basic yellow 1 are 1.0 μ g/kg,
Coffee, codeine, basic orange II, SudanⅣ are 2.5 μ g/kg.
2.4 the rate of recovery and Precision Experiment
In the chafing dish bottom flavorings of blank, chafing dish food, sweet sour pork lit-chi style, the samples such as meat are boiled in water for a while, then dress with soy, vinegar, etc., add 3 various concentrations respectively
Horizontal mixed standard solution, each pitch-based sphere replication 6 times, is quantified with extraction standard working curve, calculates average return
Yield and precision.It is 72.2%~108% to measure average recovery rate, and relative standard deviation is 2.5~9.6%, concrete outcome
It is shown in Table 4.
3 range of linearity of table, equation of linear regression, related coefficient and detection limit
TIANZHU XINGNAO Capsul experiment (n=6) in the different samples of table 4
2.5 actual samples measure
Using above test method, it have detected 30 parts of buyings and boil in water for a while, then dress with soy, vinegar, etc. meat, litchi meat with local characteristic from Putian urban district
(sweet and sour, dry deep-frying), boil in water for a while, then dress with soy, vinegar, etc. the samples such as powder, flat meat, cake, preserved fruit and chafing dish bottom flavorings, chafing dish food, detects Acid Orange II positive sample altogether
Product 3, content are 0.0159~74.2mg/kg.
The present invention is not required to using acetonitrile solution extraction object, ultra performance liquid chromatography tandem mass spectrometer measure, pre-treatment
It is simple and direct quick using the small column purifications of SPE, and high sensitivity, accuracy and precision are good, wherein poppy capsule alkaloid detection limit is excellent
In DB 31/2010-2012《The measure liquid chromatogram string of papaverine, morphine, coscopin, codeine, thebaine in chafing dish food
Join mass spectrography》In detection limit (its papaverine, morphine, coscopin, codeine, the detection limit of thebaine are 8 μ g/kg, 40 respectively
μ g/kg, 8 μ g/kg, 40 μ g/kg, 8 μ g/kg), basic orange II, the detection limit of basic flavine O are also superior to DBS22/006-2012
《The measure liquid chromatography tandem mass spectrometry of acid orange, basic orange II and basic flavine O in food》(its basic orange II and alkalescence are tender
Yellow O detection limits are 5.0 μ g/kg), it is the poppy capsule alkaloid and industry dye detected at the same time in snack products and chafing dish bottom flavorings
The effective ways of material.
The explanation of above example is only intended to help to understand method and its core concept of the invention.It should be pointed out that pair
For those skilled in the art, without departing from the principle of the present invention, the present invention can also be carried out
Some improvement and modification, these are improved and modification is also fallen into the protection domain of the claims in the present invention.
The foregoing description of the disclosed embodiments, enables professional and technical personnel in the field to realize or use the present invention.
A variety of modifications to these embodiments will be apparent for those skilled in the art, as defined herein
General Principle can be realized in other embodiments without departing from the spirit or scope of the present invention.Therefore, it is of the invention
The embodiments shown herein is not intended to be limited to, and is to fit to and the principles and novel features disclosed herein phase one
The most wide scope caused.
Claims (10)
1. a kind of ultra performance liquid chromatography tandem mass spectrometry measures 5 kinds of poppy capsule alkaloids and 6 kinds of industrial dyes in food at the same time
Method, it is characterised in that comprise the following steps：
Sample is placed in centrifuge tube by step a), adds water and acetonitrile mixes, then add anhydrous sodium acetate, shake up immediately, whirlpool
Rotation concussion 1-3min, 3-8min is centrifuged with 10000r/min, and temperature is 3-5 DEG C, takes supernatant to cross anhydrous magnesium sulfate, filtrate in
Rotation is concentrated by evaporation complete to dry, the mixed solution dissolved residue of addition 1-3mL acetonitriles and 0.1% formic acid water, solution at 35-45 DEG C
Portion is transferred in centrifuge tube, is centrifuged 1-3min with 4000r/min, is crossed the volume ratio of 0.22 μm of filter membrane, the acetonitrile and formic acid water
For 95:5；
Step b) analyzes the sample after step a processing using ultra performance liquid chromatography tandem mass spectrometry, while detects examination
The species and content of poppy capsule alkaloid and industrial dye in sample.
2. according to the method described in claim 1, it is characterized in that, 5 kinds of poppy capsule alkaloids for papaverine, morphine, can
Treat because, coscopin, thebaine.
3. according to the method described in claim 1, it is characterized in that, 6 kinds of industrial dyes for SudanⅣ, rhodamine B,
Acid Orange II, basic orange II, basic yellow 1, basic flavine O.
4. according to the method described in claim 1, it is characterized in that, in step a, the volume ratio of water and acetonitrile is 1：5.
5. according to the method described in claim 1, it is characterized in that, in step a, the sample, anhydrous sodium acetate and anhydrous sulphur
The mass ratio of sour magnesium is 2:1.5:6.
6. according to the method described in claim 1, it is characterized in that, standard solution is prepared as follows：
Appropriate standard substance is weighed respectively, and with acetonitrile or aqueous acetonitrile solution and with acetonitrile constant volume, the single mark for being configured to 100mg/L stores up
Standby liquid, then the mixed standard solution of 10mg/L is configured to, diluted step by step using preceding.
7. according to the method described in claim 1, it is characterized in that, in step b, liquid phase chromatogram condition is：
Chromatographic column：1.8 μm of SB-C18RRHD, 2.1mm × 50mm；Column temperature：35℃；Sample size：2μL；Mobile phase：A is methanol, B
For 0.1% formic acid water, gradient elution program：0-4min, 5%A, flow velocity 0.2mL/min；4.5-7min, 60%-95%A, flow velocity
0.3mL/min-0.4mL/min；7.1-11min, 95%A, flow velocity 0.5mL/min；11.5-15min 5%A, flow velocity 0.2mL/
min。
8. according to the method described in claim 1, it is characterized in that, in step b, Mass Spectrometry Conditions are：
Ion gun：Electric spray ion source, Acid Orange II use negative ion mode, remaining uses positive ion mode, negative ions
Gather at the same time；550 DEG C of ion source temperature, gas curtain gas 35psi, atomization gas 55psi, heating auxiliary gas 55psi, collision gas：
Medium, ion gun spray voltage 5500V (- 4500V)；Scan mode：Multiple-reaction monitoring.
9. according to the method described in claim 1, it is characterized in that, in step b, 5 kinds of poppy capsule alkaloids and 6 kinds of industrial dyes
Mass spectrometry parameters are as shown in table 1：
15 kinds of poppy capsule alkaloids of table and 6 kinds of industrial dye mass spectrometry parameters
10. according to the method described in claim 1, it is characterized in that, 5 kinds of poppy capsule alkaloids and 6 kinds of industrial dyes 2.5~
In the concentration range of 100 μ g/L, there is good linear relationship.
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| CN115047109B (en) * | 2022-06-17 | 2024-05-31 | 三明海关综合技术服务中心 | Method for detecting novel fentanyl mental active substances in food |
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