CN109212087A - A kind of quasi-arachidonic acid substance detection method - Google Patents
A kind of quasi-arachidonic acid substance detection method Download PDFInfo
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- 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
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Abstract
The invention discloses a kind of quasi-arachidonic acid substance detection methods, including, with volume fraction, 300 parts of water containing 0.005%BHT are added in 100 parts of sample；500 parts of methyl tertiary butyl ether(MTBE) is added, upper organic phase is shifted in centrifugation, retains lower layer's water phase；500 parts of ethyl acetate is added into lower layer's water phase, centrifugation merges upper organic phase with the upper organic phase that previous step obtains；By combined upper organic phase with after being dried with nitrogen, acetonitrile-aqueous isopropanol is used to redissolve, upper machine testing.General internal standard that the present invention selects is stable, realize accurate detection under negative ions mode, interference will not be generated to the detection of quasi-arachidonic acid substance, detection is reproducible, impurity significantly reduces in detection method, and energy can substantially reduce the pollution to instrument, the method of the present invention is convenient and efficient, extraction time is short, detection signal-to-noise ratio is obviously improved, and sensitivity greatly improves.
Description
Technical field
The invention belongs to quasi-arachidonic acid substance detection technique fields, and in particular to a kind of quasi-arachidonic acid class object
Quality detection method.
Background technique
Cyclo-oxygenase (COX-2), the enzymatics such as lipoxygenase (LOX) and cytochrome P 450 enzymes are passed through by arachidonic acid
It is metabolized the hydroxyeicosatetraenoic acid (HETEs) generated, dihydroxy eicosatetraenoic acid (DiHETEs), epoxy Eicosatetraenoic
Sour (EETs), the metabolins such as prostaglandin (PGs) and thromboxane (TX) are the main composition eicosane of eicosane acid compounds
Acid, content of these endogenous arachic acid substances in biofluid and tissue are very low.
But it low to content in this substance there are pre-treatment preparation methods at high cost, very complicated, sample at present difficult examines
The problems such as survey.Therefore how to develop it is a kind of low cost, conveniently, can be improved detection sensitivity, lower detection noise method be
The prior art has technical problem to be solved.
Summary of the invention
The purpose of this section is to summarize some aspects of the embodiment of the present invention and briefly introduce some preferable implementations
Example.It may do a little simplified or be omitted to avoid our department is made in this section and the description of the application and the title of the invention
Point, the purpose of abstract of description and denomination of invention it is fuzzy, and this simplification or omit and cannot be used for limiting the scope of the invention.
In view of above-mentioned technological deficiency, the present invention is proposed.
Therefore, as one aspect of the present invention, the present invention overcomes the deficiencies in the prior art, provides a type
Arachidonic acid-type substance detecting method.
In order to solve the above technical problems, the present invention provides the following technical scheme that a kind of quasi-arachidonic acid substance is examined
Survey method comprising,
With volume fraction, 300 parts of water containing 0.005%BHT are added in 100 parts of sample；
500 parts of methyl tertiary butyl ether(MTBE) is added, upper organic phase is shifted in centrifugation, retains lower layer's water phase；
500 parts of ethyl acetate is added into lower layer's water phase, centrifugation obtains upper organic phase and previous step
Upper organic phase merges；
By combined upper organic phase with after being dried with nitrogen, acetonitrile-aqueous isopropanol is used to redissolve, upper machine testing.
A kind of preferred embodiment as quasi-arachidonic acid substance detection method of the present invention: described to be added 500
The methyl tertiary butyl ether(MTBE) of part, centrifugation, 10000rpm is centrifuged 3min under the conditions of being included in 4 DEG C；500 are added into lower layer's water phase
The ethyl acetate of part, centrifugation, 10000rpm is centrifuged 3min under the conditions of being included in 4 DEG C.
A kind of preferred embodiment as quasi-arachidonic acid substance detection method of the present invention: described to use second
Nitrile-aqueous isopropanol redissolves, wherein with volume basis, acetonitrile: isopropanol=1:1.
A kind of preferred embodiment as quasi-arachidonic acid substance detection method of the present invention: the sample, packet
Include plasma sample.
A kind of preferred embodiment as quasi-arachidonic acid substance detection method of the present invention: further including, by 2-
Chlorophenylalanine replaces solvent as internal standard, in the detection process, is reflected by 2- chlorophenylalanine peak area and retention time
The acquisition situation of actual sample.
A kind of preferred embodiment as quasi-arachidonic acid substance detection method of the present invention: the 2- chlorobenzene third
Propylhomoserin concentration is 1 μ g/mL.
A kind of preferred embodiment as quasi-arachidonic acid substance detection method of the present invention: further including, and uses
4,4 '-di-2-ethylhexylphosphine oxides (2- chloroaniline), P-anisidine, l-tyrosine methyl esters, 3- chloroaniline, 2,4- dimethyl quinoline, sulfanilamide (SN) pyrrole
Pyridine, Atrazine, sulfadoxine, DL-leucine, N- benzoyl-l-tyrosine ethyl ester, 6- phenyl -2- deracil, N-
(toluoyl) glycine, Metronidazole, enoxolone, flavanones, 6-caprolactone, 2-aminopyridine
Mixed sample as general quality-control product, detection is analyzed according to spectrogram overlapping cases and is examined at least into the general quality-control product of two needles every time
Survey the state of instrument.
A kind of preferred embodiment as quasi-arachidonic acid substance detection method of the present invention: by quality ratio,
Described 4,4 '-di-2-ethylhexylphosphine oxides (2- chloroaniline): P-anisidine: l-tyrosine methyl esters: 3- chloroaniline: 2,4- dimethyl quinolines: sulphur
Amine pyridine: Atrazine: sulfadoxine: DL-leucine: N- benzoyl-l-tyrosine ethyl ester: 6- phenyl -2- deracil:
N- (toluoyl) glycine: Metronidazole: enoxolone: flavanones: 6-caprolactone: 2- amino pyrrole
Pyridine=1:0.5:0.5:2:1:0.4:0.1:0.1:0.2:0.5:0.5:1:1:0.1:0.5:2:0.2.
A kind of preferred embodiment as quasi-arachidonic acid substance detection method of the present invention: described 4,4 '-is sub-
The concentration of methyl bis- (2- chloroanilines) is 1 μ g/mL, the concentration of P-anisidine is 0.5 μ g/mL, the concentration of l-tyrosine methyl esters is
0.5 μ g/mL, 3- chloroaniline concentration be 2 μ g/mL, 2,4- dimethyl quinoline concentration be 1 μ g/mL, the concentration of sulfapryidine is
0.4 μ g/mL, Atrazine concentration be 0.1 μ g/mL, the concentration of sulfadoxine is 0.1 μ g/mL, the concentration of DL-leucine is
0.2 μ g/mL, N- benzoyl-l-tyrosine ethyl ester concentration are 0.5 μ g/mL, the concentration of 6- phenyl -2- deracil is 0.5
μ g/mL, N- (toluoyl) glycine concentration be 1 μ g/mL, the concentration of Metronidazole is 1 μ g/
ML, enoxolone concentration be 0.1 μ g/mL, the concentration of flavanones is 0.5 μ g/mL, the concentration of 6-caprolactone is 2 μ g/mL, 2-
The concentration of aminopyridine is 0.2 μ g/mL.
Beneficial effects of the present invention: the method for the present invention significantly improves the detection sensitivity of the substance.The present invention is in reality
Quality-control product need not be individually prepared during testing, especially when mass detection, the general quality-control product of the present invention is particularly convenient, can
Going out the unstable reason of instrument according to spectrogram overlapping cases successful analysis, accuracy of judgement degree reaches 100%, meanwhile, present invention choosing
2- chlorophenylalanine is selected as general internal standard, accurate to monitor sample collection state, the general internal standard that the present invention selects is stable, positive and negative
Realize that accurate detection, will not to generate interference, detection to the detection of quasi-arachidonic acid substance reproducible under ion mode,
Impurity significantly reduces in detection method, and can substantially reduce the pollution to instrument, and the method for the present invention is convenient and efficient, mentions
Take that the time is short, detection signal-to-noise ratio is obviously improved, sensitivity greatly improves.
Detailed description of the invention
In order to illustrate the technical solution of the embodiments of the present invention more clearly, required use in being described below to embodiment
Attached drawing be briefly described, it should be apparent that, drawings in the following description are only some embodiments of the invention, for this
For the those of ordinary skill of field, without any creative labor, it can also be obtained according to these attached drawings other
Attached drawing.Wherein:
Fig. 1 is the setting demonstration of the method for the present invention MultiQuant software parameter.
Fig. 2 is the datagram for the sample internal standard exception that the present invention detects.
Fig. 3 is the peak figure out for the exceptional sample that the present invention detects.
Fig. 4 is the peak figure out that the present invention resurveys rear normal sample.
Fig. 5 is quality-control product of the present invention appearance situation map on liquid chromatograph-mass spectrometer device.
Fig. 6 is overlap of peaks situation map of the quality-control product of the present invention on liquid chromatograph-mass spectrometer device.
Fig. 7 is the appearance and overlap of peaks situation map of the mixed mark quality-control product of the present invention in one-time authentication experiment.
Fig. 8 is in one-time authentication experiment, it is known that the appearance and overlap of peaks situation map of sample.
Fig. 9 is the appearance situation map of the mixed mark quality-control product of the present invention in one-time authentication experiment.
Figure 10 is in one-time authentication experiment, it is known that the appearance and overlap of peaks situation map of sample.
Figure 11 is the appearance and overlap of peaks situation map of the mixed mark quality-control product of the present invention in one-time authentication experiment.
Figure 12 is in one-time authentication experiment, it is known that the appearance and overlap of peaks situation map of sample.
Specific embodiment
In order to make the foregoing objectives, features and advantages of the present invention clearer and more comprehensible, right combined with specific embodiments below
A specific embodiment of the invention is described in detail.
In the following description, numerous specific details are set forth in order to facilitate a full understanding of the present invention, but the present invention can be with
Implemented using other than the one described here other way, those skilled in the art can be without prejudice to intension of the present invention
In the case of do similar popularization, therefore the present invention is not limited by the specific embodiments disclosed below.
Secondly, " one embodiment " or " embodiment " referred to herein, which refers to, may be included at least one realization side of the invention
A particular feature, structure, or characteristic in formula." in one embodiment " that different places occur in the present specification not refers both to
The same embodiment, nor the individual or selective embodiment mutually exclusive with other embodiments.
Embodiment 1:
Using the triple level four bars of ultra performance liquid chromatography -/linear ion hydrazine tandem mass spectrum (UPLC-QTRAP) technology, matter
Spectrometer is AB Sciex Q-TRAP 6500, and analysis software is Multi Quant.
It takes the plasma sample of 100uL in EP pipe, adds the water of 300uL (containing 0.005%BHT)；
Into EP pipe be added 500uL MTBE (methyl tertiary butyl ether(MTBE)), be vortexed mix, under the conditions of 4 DEG C 10000rpm from
It after heart 3min, is transferred to upper layer is organic in new EP pipe, retains lower layer's water phase；
The ethyl acetate of 500uL is added into the water phase of lower layer, is vortexed after mixing, 10000rpm is centrifuged under the conditions of 4 DEG C
After 3min, upper organic phase is merged with the upper organic phase that previous step obtains；
By combined upper organic phase with after being dried with nitrogen, is redissolved, is packed into sample bottle using acetonitrile/isopropanol (1:1),
To upper machine testing.
Simultaneously as quasi-arachidonic acid substance content is lower in plasma sample, therefore by extracting method, sample when detection
Product acquisition situation, instrument fluctuation etc. are affected, and for the monitoring of sample acquisition situation, the present invention selects 2- chlorophenylalanine to make
For internal standard, it is configured to the internal standard extracting solution of 1 μ g/mL, replaces pure solvent to experiment sample with internal standard 2- chlorophenylalanine extracting solution
It extracts, guarantees that interior target concentration is consistent in each sample, upper machine acquisition data pass through in data acquisition
The setting of MultiQuant software parameter, checks internal standard peak area and retention time, acquires situation by interior target, reflects practical sample
This acquisition situation, individual samples of internal standard exception, then open corresponding acquisition data and check, data exception adopt again.
The setting of MultiQuant software parameter: the other materials in addition to internal standard are deleted；Generate target knot in all samples
Fruit table；It chooses area one to arrange, clicks creates a metric plot, generate area line chart, line chart ordinate is changed
At the form of percentage, addition one represents internal standard area average value horizontal line, sample internal standard area 10% model above and below average value
In enclosing, it is believed that sample data acquisition is normal, returns again to data acquisition software beyond 10% and checks sample collection data, exceptional sample
Resubmit acquisition；It chooses retention time one to arrange, clicks creates a metric plot, generate retention time line chart, partially
Difference is more than that 0.1 returned data acquisition software checks sample collection data, and exceptional sample resubmits acquisition.
The present invention passes through repeated screening from numerous substances, selects 2- chlorophenylalanine as internal standard, finds it positive and negative
Under ion mode can appearance and peak type it is good, and be not easy to test substance generate interference, can using the method for the present invention
The acquisition situation of accurate measurements sample, and will not omit, for example, when detection sample mixes bubble during sample-adding,
It will cause sample volume reduction, cause out peak response reduction, so that the quantitative inaccuracy of the final sample, art methods
This technical problem can not be monitored, after internal standard quality-control product of the present invention, when a certain sample mixes bubble, is added in the sample
The interior target appearance response entered can also reduce, therefore can accurately find the out of condition sample of the acquisition.
And works as and select other substances as internal standard, such as 3- iodotyrosine, 15 fluorine octanoic acids, Sucralose, 3- chlorobenzene
Whens amine, tridecanoic acid, glycine, benzoic acid, succinic acid etc., due to influencing difference too under its is unstable or negative ions mode
Greatly or sample material to be also easy to produce interference, detection repeatability bad, therefore be not suitable as general internal standard.
Fig. 1 is the setting of the method for the present invention MultiQuant software parameter.Fig. 2 is the sample of internal standard exception of the present invention, and Fig. 3 is
The sample goes out peak figure, from Fig. 2 and Fig. 3 it is found that when internal standard is abnormal, illustrates sample collection exception, Fig. 4 is by the sample
Go out peak figure after resurveying sample introduction, comparison diagram 3 illustrates choosing of the present invention it is found that peak response is decreased obviously out when Fig. 3 sample collection
2- chlorophenylalanine, which is selected, as internal standard can accurately reflect sample collection situation.
In order to the fluctuation status of instrument when monitor and detection simultaneously, the present invention screens 17 mixed marks and forms general Quality Control
Product, 17 standard specimens of general quality-control product of the invention and its are shown in Table 1 using final concentration:
Table 1
The present invention selects the compound that separating degree is good, response is suitable, stable and is used as mixed mark, 17 marks by testing repeatedly
Sample appearance time is substantially uniformly distributed in 0~11 minute, mutually smaller on the influence of appearance time each other, is responded in 1*10^6~3*
Between 10^7, belonging to the best response range of used test instrument, 17 standard specimen compound structures are stablized, and it is not volatile, rotten, two
Each substance responds fluctuation is prepared within 20% using hundreds of potential possibility as the substance of standard items respectively between secondary acquisition
At the mother liquor of 1mg/mL and the working solution of 1 μ g/mL, is developed with the liquid matter method that working solution carries out standard items, determines testing conditions,
Again with mother liquor at a hybrid standard product solution, upper machine acquisition, and the concentration of standard items is adjusted, is made into mixed mark, upper machine is adopted
Collect data, causes quality-control product unstable or the detection peak of each standard specimen due to that may react to each other between the standard specimen of various combination
Between be easy to interfere with each other or respond bad etc., therefore the present invention is by combining repeatedly, screening, it is final determine 17 can
Suitable for the mixed mark quality-control product of various different classes of substance detections, 17 mixed mark quality-control product chromatographies go out peak figure as shown in figure 5, from figure
17 standard specimen detection peaks are separated from each other known to 5, are not interacted substantially, and respond well, in the most suitable range of instrument, 17
A mixed mark total ion chromatogram overlay chart is as shown in Figure 6.The mixed mark quality-control product overlapping of two needles is fine as can be seen from Figure 6, peak height
Unanimously, show that instrument response is stablized, retention time is consistent, shows that chromatographic isolation situation is stablized, instrument state is good.Matter of the present invention
Control product can be used as general quality-control product, the detection quality-control product suitable for all substances.
Judge that instrument fluctuates by the data total ion chromatogram overlapping cases that front and back acquires in data acquisition
Situation, chromatogram overlapping preferably illustrate instrument stabilizer, and overlapping is bad then to judge instrument state by overlapping cases, such as when reservation
Between be not overlapped explanation may be column pressure or column temperature it is unstable, peak height be overlapped not Shang explanation may be chromatographic column block or mass spectrum dirt
Dye etc..
The mixed mark quality-control product of the present invention is sufficiently stable, and prepares simply, and dosage is few, at low cost.The present invention passes through for difference
The multiplicating of kind of substance, which detects, to be proved, being capable of accurately reaction chromatography instrument or mass spectrometric instrument using quality-control product of the present invention
State (whether instrument stable, column pressure, whether column temperature is stable, whether pillar has blocking, whether has pollution etc.) and sample to be tested
State, and the unstable reason of instrument can be gone out according to spectrogram overlapping cases successful analysis, accuracy of judgement degree reaches 100%.
Using the known sample of different conditions as standard, to verify quality-control product of the invention for different sample states and instrument
The monitoring accuracy of device state, each known sample do primary repetition sample introduction, each mixed mark quality-control product, which is done, once repeats sample introduction,
If the appearance and overlapping cases of known sample are consistent with the appearance of the mixed mark quality-control product of the present invention and overlapping cases, illustrate the present invention
The case where mixed mark quality-control product is for sample and instrument state reflection is accurate.It is verified by many experiments, finds the mixed mark matter of the present invention
Control product can 100% reflected sample acquisition situation and when sampling instrument operating status, by the appearance of mixed mark quality-control product and again
Folded situation, can reflect the acquisition situation of sample to be tested.
As shown in Figure 7 and Figure 8, Fig. 7 is the appearance and overlap of peaks feelings of the mixed mark quality-control product of the present invention in one-time authentication experiment
Condition, as shown in fig. 7, the mixed mark quality-control product overlapping of two needles is very well, peak height is consistent, shows that instrument response is stablized, retention time is consistent, table
Bright chromatographic isolation situation is stablized, and instrument state is good, as shown in figure 8, in this confirmatory experiment, it is known that the appearance of sample and again
It is folded same fine, illustrate that the mixed mark quality-control product of the present invention is capable of the normal condition of accurate reaction kit.
As shown in Figure 9 and Figure 10, Fig. 9 is the appearance situation of the mixed mark quality-control product of the present invention, from figure in one-time authentication experiment
9 find out, quality-control product peak type is bad, separation is bad, and the mixed mark non-appearance of quality-control product of 10min or so, illustrate chromatographic apparatus when detection
There are problems.Figure 10 is in this confirmatory experiment, it is known that the appearance and overlapping cases of sample, as can be seen from Figure 10, it is known that sample is same
Sample separation is bad, and 8~10min is responded and is lower, and quality-control product of the present invention and known sample response are consistent, and by verifying,
There is problem when detecting in chromatograph.Illustrate that the mixed mark quality-control product of the present invention is capable of the abnormality of accurate reaction kit.
As is illustrated by figs. 11 and 12, Figure 11 is the appearance and peak weight of the mixed mark quality-control product of the present invention in one-time authentication experiment
Folded situation, as shown in figure 11, the mixed mark quality-control product overlap of peaks of two needles are bad, and the offset of 1~3min retention time illustrates to supervise time keeping instrument
Unstable, Figure 12 is in this confirmatory experiment, it is known that the appearance and overlapping cases of sample, as can be seen from Figure 12, it is known that sample is same
Overlap of peaks is bad, and 1~3min sample retention time equally exists offset, and by verifying, and column pressure is not when detecting for chromatograph
Surely, illustrate that the mixed mark quality-control product of the present invention is capable of the abnormality of accurate reaction kit.
Embodiment 2 (reference examples):
It takes the sample of 100uL in EP pipe, adds the water of 300uL (containing 0.005%BHT)；
Into EP pipe be added 500uL MTBE (methyl tertiary butyl ether(MTBE)), be vortexed mix, under the conditions of 4 DEG C 10000rpm from
It after heart 3min, is transferred to upper layer is organic in new EP pipe, retains lower layer's water phase；
The MTBE (methyl tertiary butyl ether(MTBE)) of 500uL is added into the water phase of lower layer, is vortexed after mixing, under the conditions of 4 DEG C
After 10000rpm is centrifuged 3min, upper organic phase is merged with the upper organic phase that previous step obtains；
By combined upper organic phase with after being dried with nitrogen, is redissolved, is packed into sample bottle using acetonitrile/isopropanol (1:1),
To upper machine testing.
Embodiment 3 (reference examples):
It takes the sample of 100uL in EP pipe, adds the water of 300uL (containing 0.005%BHT)；
The ethyl acetate of 500uL is added into EP pipe, is vortexed and mixes, it, will after 10000rpm is centrifuged 3min under the conditions of 4 DEG C
Upper layer is organic to be transferred in new EP pipe, and lower layer's water phase is retained；
The ethyl acetate of 500uL is added into the water phase of lower layer, is vortexed after mixing, 10000rpm is centrifuged under the conditions of 4 DEG C
After 3min, upper organic phase is merged with the upper organic phase that previous step obtains；
By combined upper organic phase with after being dried with nitrogen, is redissolved, is packed into sample bottle using acetonitrile/isopropanol (1:1),
To upper machine testing.
2 Examples 1 to 3 extracting method of table extracts the comparison of quasi-arachidonic acid substance signal-to-noise ratio
A variety of arachidonic acid-type compounds are detected simultaneously, are compared by ethyl acetate with the combination of two kinds of solvents of MTBE
The extraction signal-to-noise ratio of single solvent is obviously improved, and the method for the present invention significantly improves the detection sensitivity of the substance.The present invention
Quality-control product need not be individually prepared during the experiment, especially when mass detection, the general quality-control product of the present invention is particularly convenient,
It can go out the unstable reason of instrument according to spectrogram overlapping cases successful analysis, accuracy of judgement degree reaches 100%, meanwhile, this hair
It is bright to select 2- chlorophenylalanine as general internal standard, it is accurate to monitor sample collection state, the general internal standard that the present invention selects is stable,
Accurate detection is realized under negative ions mode, interference, detection repetition will not be generated to the detection of quasi-arachidonic acid substance
Property it is good, impurity significantly reduces in detection method, and can substantially reduce pollution to instrument, and the method for the present invention facilitates fast
It is prompt, extraction time is short, detection signal-to-noise ratio is obviously improved, sensitivity greatly improves.
It should be noted that the above examples are only used to illustrate the technical scheme of the present invention and are not limiting, although referring to preferable
Embodiment describes the invention in detail, those skilled in the art should understand that, it can be to technology of the invention
Scheme is modified or replaced equivalently, and without departing from the spirit and scope of the technical solution of the present invention, should all be covered in this hair
In bright scope of the claims.
Claims (9)
1. a kind of quasi-arachidonic acid substance detection method, it is characterised in that: including,
With volume fraction, 300 parts of water containing 0.005%BHT are added in 100 parts of sample；
500 parts of methyl tertiary butyl ether(MTBE) is added, upper organic phase is shifted in centrifugation, retains lower layer's water phase；
500 parts of ethyl acetate, centrifugation, the upper layer that upper organic phase and previous step are obtained are added into lower layer's water phase
Organic phase merges；
By combined upper organic phase with after being dried with nitrogen, acetonitrile-aqueous isopropanol is used to redissolve, upper machine testing.
2. quasi-arachidonic acid substance detection method as described in claim 1, it is characterised in that: 500 parts of the addition
Methyl tertiary butyl ether(MTBE), centrifugation, 10000rpm is centrifuged 3min under the conditions of being included in 4 DEG C；500 parts are added into lower layer's water phase
Ethyl acetate, centrifugation, 10000rpm is centrifuged 3min under the conditions of being included in 4 DEG C.
3. quasi-arachidonic acid substance detection method as claimed in claim 1 or 2, it is characterised in that: described to use acetonitrile-
Aqueous isopropanol redissolves, wherein with volume basis, acetonitrile: isopropanol=1:1.
4. quasi-arachidonic acid substance detection method as claimed in claim 1 or 2, it is characterised in that: the sample, including
Plasma sample.
5. quasi-arachidonic acid substance detection method as claimed in claim 1 or 2, it is characterised in that: further include, by 2- chlorine
Phenylalanine replaces solvent as internal standard, in the detection process, is reflected by 2- chlorophenylalanine peak area and retention time real
The acquisition situation of border sample.
6. quasi-arachidonic acid substance detection method as claimed in claim 5, it is characterised in that: the 2- chlorophenylalanine
Concentration is 1 μ g/mL.
7. the quasi-arachidonic acid substance detection method as described in any in claim 1,2,6, it is characterised in that: also wrap
Include, using 4,4 '-di-2-ethylhexylphosphine oxides (2- chloroaniline), P-anisidine, l-tyrosine methyl esters, 3- chloroaniline, 2,4- dimethyl quinoline,
Sulfapryidine, Atrazine, sulfadoxine, DL-leucine, N- benzoyl-l-tyrosine ethyl ester, 6- phenyl -2- thiocarbamide are phonetic
Pyridine, N- (toluoyl) glycine, Metronidazole, enoxolone, flavanones, 6-caprolactone, 2- ammonia
The mixed sample of yl pyridines is as general quality-control product, and detection is at least into the general quality-control product of two needles every time, according to spectrogram overlapping cases
The state of analysis and detecting instrument.
8. quasi-arachidonic acid substance detection method as claimed in claim 7, it is characterised in that: by quality ratio, described
4,4 '-di-2-ethylhexylphosphine oxides (2- chloroaniline): P-anisidine: l-tyrosine methyl esters: 3- chloroaniline: 2,4- dimethyl quinolines: sulfanilamide (SN) pyrrole
Pyridine: Atrazine: sulfadoxine: DL-leucine: N- benzoyl-l-tyrosine ethyl ester: 6- phenyl -2- deracil: N-
(toluoyl) glycine: Metronidazole: enoxolone: flavanones: 6-caprolactone: 2-aminopyridine
=1:0.5:0.5:2:1:0.4:0.1:0.1:0.2:0.5:0.5:1:1:0.1:0.5:2:0.2.
9. quasi-arachidonic acid substance detection method as claimed in claim 8, it is characterised in that: described 4,4 '-methylene
The concentration of bis- (2- chloroanilines) is 1 μ g/mL, the concentration of P-anisidine is 0.5 μ g/mL, the concentration of l-tyrosine methyl esters is 0.5 μ
The concentration of g/mL, 3- chloroaniline is 2 μ g/mL, the concentration of 2,4- dimethyl quinoline is 1 μ g/mL, the concentration of sulfapryidine is 0.4 μ
G/mL, Atrazine concentration be 0.1 μ g/mL, the concentration of sulfadoxine is 0.1 μ g/mL, the concentration of DL-leucine is 0.2 μ
G/mL, N- benzoyl-l-tyrosine ethyl ester concentration are 0.5 μ g/mL, the concentration of 6- phenyl -2- deracil is 0.5 μ g/
The concentration of mL, N- (toluoyl) glycine is 1 μ g/mL, the concentration of Metronidazole is 1 μ g/mL, sweet
The concentration of careless hypo acid is 0.1 μ g/mL, the concentration of flavanones is 0.5 μ g/mL, the concentration of 6-caprolactone is 2 μ g/mL, 2- amino pyrrole
The concentration of pyridine is 0.2 μ g/mL.
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