CN106124645A - Needle-based Solid-Phase Extraction filter - Google Patents
Needle-based Solid-Phase Extraction filter Download PDFInfo
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- CN106124645A CN106124645A CN201610409291.9A CN201610409291A CN106124645A CN 106124645 A CN106124645 A CN 106124645A CN 201610409291 A CN201610409291 A CN 201610409291A CN 106124645 A CN106124645 A CN 106124645A
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- phase extraction
- needle
- filter
- solid
- sieve plate
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- 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
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- 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
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- 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
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
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- 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
- G01N2030/062—Preparation extracting sample from raw material
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
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Abstract
The invention discloses a kind of needle-based Solid-Phase Extraction filter, including housing, be respectively provided with the syringe nipple interface and pintle interface being connected with housing cavity in the both sides up and down of housing;Upper sieve plate, solid phase extraction filler, lower sieve plate and filter membrane is set gradually from top to bottom in housing cavity;The sieve aperture that aperture is 95~105 μm it is respectively equipped with on upper sieve plate and lower sieve plate.The particle diameter of solid phase extraction filler is 110~130 μm.Solid-Phase Extraction is combined in one apparatus with membrane filtration by the present invention first, is greatly simplified operating procedure, saves solvent, filler and running cost;Present invention can be suitably applied to different samples and solvent, applied widely, effective.
Description
Technical field
The present invention relates to material separation technology field, particularly relate to a kind of needle-based Solid-Phase Extraction filter.
Background technology
Sample pre-treatments before chromatography is it is critical that step, current Solid-Phase Extraction (Solid-phase
Extraction, SPE) it is highly important method, target components enrichment method can be reached the purpose of Sample Purification on Single.With biography
System liquid-liquid extraction is compared, and can improve the response rate, more effectively target components be separated with impurity chaff interference, is widely used in doctor
The fields such as medicine, food, environment, chemical industry.
Bulky grain analytical tool detections such as liquid chromatograph of being allowed for access need to be removed through the component of Solid-Phase Extraction eluting to analyze.Filter
Membrance separation is the most noticeable in application biological, chemical analysis field.Structure of pinhead type filter is simple, with low cost, and
And good filtration effect, general applicability is strong, is widely used in the lab, as the filtration treatment device of small samples.
The structure of current existing syringe-driven filter is: arrange filter membrane in housing cavity, in the both sides up and down of housing
It is respectively provided with the syringe nipple interface and pintle interface being connected with housing cavity, the most only does filtration use, do not possess solid phase
Extraction function.
Solid-Phase Extraction and the separate operations of membrane filtration, add the complexity of consumables cost and pre-treatment.Prior art
Solid-Phase Extraction column volume is relatively big, consumes filler and solvent is more, and extracting operation is relatively costly.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of simple in construction, needle-based Solid-Phase Extraction easy to use filters
Device.
In order to solve above-mentioned technical problem, the present invention provides a kind of needle-based Solid-Phase Extraction filter, including housing, at shell
The both sides up and down of body are respectively provided with the syringe nipple interface and pintle interface being connected with housing cavity;
Upper sieve plate, solid phase extraction filler, lower sieve plate and filter membrane is set gradually from top to bottom in housing cavity;
The sieve aperture that aperture is 95~105 μm (preferably 100 μm) it is respectively equipped with on described upper sieve plate and lower sieve plate.
Improvement as the needle-based Solid-Phase Extraction filter of the present invention: the particle diameter of described solid phase extraction filler be 110~
130 μm (preferably 120 μm), solid phase extraction filler is-C18 ,-C8, silica gel, aluminium oxide, Florisil, kieselguhr, activated carbon,
At least one in Diol (glycol-based).
Further improvement as the needle-based Solid-Phase Extraction filter of the present invention: filter membrane is nylon.
Further improvement as the needle-based Solid-Phase Extraction filter of the present invention: the pore size filter of filter membrane is 0.5 μm.
Further improvement as the needle-based Solid-Phase Extraction filter of the present invention: syringe nipple interface and pintle interface
It is standard straight socket or standard thread mouth.
When the present invention is actually used, syringe nipple interface is connected with standard syringe nipple, pintle interface and injection
Device pintle is connected.
The needle-based Solid-Phase Extraction filter of the present invention, is Solid-Phase Extraction to be effectively combined with membrane filtration, solves existing
Solid-Phase Extraction present in technology, membrane filtration step are loaded down with trivial details, conventional solid extraction column volume compared with big, use that filler is more, eluting
Liquid consumes the problems such as more, running cost is higher.
The using method of the needle-based Solid-Phase Extraction filter of the present invention, is specially and follows the steps below successively:
One, prepared by sample: accurate weighing sample, extracts with aqueous solutions of organic solvent, takes supernatant after high speed centrifugation 15min
Liquid, repeats precipitate to extract operation 2 times, merges supernatant evaporated under reduced pressure, adds a small amount of aqueous solutions of organic solvent and redissolves.
Two, filler activator: use aqueous solutions of organic solvent that packing layer (solid phase extraction filler) is carried out activation processing, use
Aqueous solutions of organic solvent is injected into housing cavity from syringe nipple interface by 3mL syringe, and aqueous solutions of organic solvent flows successively
After upper sieve plate, solid phase extraction filler, lower sieve plate and filter membrane, it is discharged housing from pintle seam.Aqueous solutions of organic solvent
The speed flowing through packing layer is about 0.1~2mL/min.Effluent collected and abandons, reaching needle-based Solid-Phase Extraction filter
Activation and the purpose cleaned.
Three, loading: taking 3mL syringe extraction appropriate amount of sample solution, the speed syringe needle formula with 0.1~2mL/min is solid
Phase extraction filter, target components is attracted on filler, and impurity directly flows out needle-based Solid-Phase Extraction with sample solution and filters
Device also abandons.
Four, drip washing: inject 3mL leacheate with the speed of 0.1~2mL/min to needle-based Solid-Phase Extraction filter, described
Leacheate be ultra-pure water (containing 0~10% methanol or acetonitrile) in order to wash away the impurity co-adsorption with target components.
Five, eluting: adding 1~3mL eluant to needle-based Solid-Phase Extraction filter, described eluant is methanol or acetonitrile
(containing 0~30% ultra-pure water), flow velocity is 0.1~2mL/min, collects eluent.
During invention, inventor has found that syringe-driven filter and solid-phase extraction column all can produce in experimental implementation
Raw relatively strong pressure, can cause the problem that solution is detained after both pressure superposition;Meanwhile, filtration experiment solvent is single, and solid phase extraction
It is complicated that treating excess syndrome tests dicyandiamide solution, can cause the problem that filter membrane dissolves;The two reason makes Solid-Phase Extraction in the combination with filter membrane
Difficulty, does not the most temporarily have Related product.Therefore, the present invention use particle diameter be 110~130 μm fillers (such as ,-
C18, when particle diameter is 120 μm, specific surface area about 500m2/ g), make solvent under normal pressure can naturally flow out solid phase extraction filler, filter membrane
Material uses nylon, compares traditional teflon membrane filter (being only applicable to organic solvent) and mixed cellulose ester is (the most applicable
In water), nylon membrane is with inorganic and organic solvent is the most compatible;Filter sizes uses 0.5 μm, compares conventional 0.45 and 0.22 μm filter
Film pressure is less.The present invention combines above technological improvement point makes Solid-Phase Extraction be combined with microporous filter membrane.
The present invention has a following technical advantage:
1, first Solid-Phase Extraction is combined in one apparatus with membrane filtration, is greatly simplified operating procedure, save solvent, fill out
Material and running cost.
2, follow sample prepare, filler activator, loading, drip washing and elution step, simple easily training, and without specific apparatus
Equipment, practical, applicable laboratory is widely used.
3, the method for the present invention is stable, efficient, by regulating different fillers and membrane material, is applicable to different samples
With solvent, applied widely, effective.
Accompanying drawing explanation
Below in conjunction with the accompanying drawings the detailed description of the invention of the present invention is described in further detail.
Fig. 1 is the cross-sectional view of the needle-based Solid-Phase Extraction filter of embodiment 1;
Wherein, 1, upper sieve plate;2, solid phase extraction filler;3, lower sieve plate;4, filter membrane;5, housing;6, syringe nipple interface;
7, pintle interface.
Fig. 2 is to use needle-based Solid-Phase Extraction filter to carry out the PHOSPHATIDYL ETHANOLAMINE of pre-treatment and the color of phosphatidylcholine
Spectrogram.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is described further, but protection scope of the present invention is not limited in
This: below in conjunction with accompanying drawing describe in detail the present invention detailed description of the invention, make those skilled in the art be more clearly understood that as
What puts into practice the present invention.Should be appreciated that although combining its preferred embodiment describes the present invention, but these embodiments
Simply illustrate rather than limit the scope of the present invention.
Embodiment 1, a kind of needle-based Solid-Phase Extraction filter, as it is shown in figure 1, include housing 5, at up and down the two of housing 5
Side is respectively provided with the syringe nipple interface 6 with housing 5 intracavity inter-connection and pintle interface 7;In housing 5 inner chamber from top to bottom
Set gradually upper sieve plate 1, solid phase extraction filler 2, lower sieve plate 3 and filter membrane 4;That is, upper sieve plate 1, solid phase extraction filler 2, lower sieve plate 3
It is stacked layer by layer for gapless with filter membrane 4.
The sieve aperture that aperture is 100 μm it is respectively provided with on upper sieve plate 1, lower sieve plate 3.Upper sieve plate 1 is just to syringe nipple interface
6, and filter membrane 4 is just to pintle interface 7.Syringe nipple interface 6 and pintle interface 7 are standard straight socket or standard thread mouth.
This octadecyl chlorosilane C18 be particle diameter be the filler of 120 μm, specific surface area about 500m2/g。
Time actually used, following size can be taked:
Upper sieve plate 1, lower sieve plate 3 are polythene PE material (thickness 1mm), and solid phase extraction filler 2 is octadecyl chloride silicon
Alkane C18 (thickness 2.5mm), filter membrane 4 are nylon material (thickness about 0.5mm, aperture 0.5 μm).
The external dimensions of whole needle-based Solid-Phase Extraction filter is: diameter 15mm, thickness 10m (disregard interface).
The diameter of upper sieve plate 1, solid phase extraction filler 2, lower sieve plate 3 and filter membrane 4 all=housing 5 internal diameter (13mm).
Phosphatidylcholine and phosphatidyl in experiment 1, the needle-based Solid-Phase Extraction filter prawn head tissue of Application Example 1
The enrichment of ethanolamine purifies.
One, prepared by sample: accurately weighed ground prawn head sample 0.1g, mixes with chloroform-methanol (2:1, volume ratio)
Liquid 3mL ice-bath ultrasonic extracts 30min, adds ultra-pure water 1mL and with 12,000rpm/min high speed frozen centrifugation 5 minutes.Use
The lower clear liquid of 200mL liquid-transfering gun transfer, adds 2mL chloroform in remaining supernatant and solid content, repeats to carry according to aforesaid operations
Take twice.Merge lower clear liquid and dry up with nitrogen, adding 1mL methanol and redissolve, obtain crude extract.
Two, filler activator: needle-based Solid-Phase Extraction filter uses 3mL methanol, 50% methanol water-soluble before using the most respectively
Liquid, acetonitrile activate and balance.
Three, loading: take loading after crude extract 100 μ L with liquid-transfering gun, and with 1mL/min flow velocity by needle-based Solid-Phase Extraction
Filter.
Four, drip washing: inject 3mL leacheate (5% acetonitrile water with the speed of 1mL/min to needle-based Solid-Phase Extraction filter
Solution), wash away the impurity co-adsorption with phosphatidylcholine and PHOSPHATIDYL ETHANOLAMINE.
Five, eluting: carry out eluting with 90% acetonitrile 1mL, flow velocity is 1mL/min, dries up with nitrogen after collecting eluent, and
Mass spectral analysis is prepared with 200 μ L acetonitriles after redissolving.
Six, liquid chromatography/mass spectrometry detection: use Agilent 1100 liquid chromatographic system, selects YMC Triart diol
HILIC chromatographic column (4.6mm × 250mm, 3 μm);Flowing is A mutually: containing the acetonitrile solution of 53mM formic acid, B: the formic acid containing 53mM is water-soluble
Liquid;Flow velocity is 200 μ L/min;Eluent gradient system is 0 15min:2%A (98%B), 15 60min: from 2%A to 40%
A, 60 70min:40%A, 70 80min: from 40%A to 2%A.Mass spectrum uses the QSTAR triple four of American AB Sciex company
Level bar/flight time mass spectrum, sweep limits is from m/z 450 to 950;Under negative ions pattern, ion spray voltage is respectively 5.5
And 4.5kV.Ion source temperature is 450 DEG C;Atomization gas and gas curtain edema caused by disorder of QI are not 35 and 20psi.Bunch voltage and collision voltage is gone to divide
It is not 100 and 40V.The spectrogram of phosphatidylcholine and PHOSPHATIDYL ETHANOLAMINE is as shown in Figure 2.
Result
This experiment carries out gradient dilution sample introduction (that is, with this alternative steps one with standard concentration scope 0.5-200 μ g/mL
Crude extract), standard substance content is abscissa, and corresponding peak area is vertical coordinate, with y=a+bx linear fit, phosphatidyl gallbladder
Alkali and the R of PHOSPHATIDYL ETHANOLAMINE2It is all higher than 0.99.The detection of target analytes is determined respectively with 3 times and 10 times of signal to noise ratios (S/N)
Limit (LOD) and quantitative limit (LOQ), the detection limit recording phosphatidylcholine and PHOSPHATIDYL ETHANOLAMINE is respectively 0.12 and 0.23 μ g/
ML, is quantitatively limited to 0.37 and 0.71 μ g/mL.Blank muscle tissue sample pitch-based sphere is respectively LOQ and 20 times of LOQ, each
Concentration arranges 3 samples, and by drafting determination of experimental method, calculating precision, the response rate are shown in Table 1.Precision is less than 6.1%, table
Bright instrument precision is good;The phospholipid response rate is 82.3%~90.7%, meets and analyzes test needs.
The quantitative parameter of table 1, phosphatidylcholine and PHOSPHATIDYL ETHANOLAMINE
As fully visible, the needle-based Solid-Phase Extraction filter of the present invention can effectively the phosphatidylcholine in prawn head and
PHOSPHATIDYL ETHANOLAMINE enrichment purifies, single needle-based Solid-Phase Extraction filter with low cost, and the response rate is high, precision
Good.Compared with existing commercialization solid-phase extraction column, Solid-Phase Extraction is combined in one apparatus with membrane filtration, is greatly simplified behaviour
Make step, save solvent, filler and running cost.
Embodiment 2, made the solid phase extraction filler 2 in embodiment 1 into octyl silicon respectively by octadecyl chlorosilane C18
Alkane bonded silica gel C8, silica gel, aluminium oxide, Florisil, kieselguhr, activated carbon, Diol (glycol-based);Particle diameter is still 120 μm, its
More than be equal to embodiment 1.
Detect as tested 1 with the needle-based Solid-Phase Extraction filter of gained, phosphatidylcholine and phosphatidyl ethanol
The quantitative parameter of amine is as described in Table 2.
Table 2
Comparative example 1, by the octadecyl chlorosilane C18 in embodiment 1 by " particle diameter 120 μm, specific surface area about 500m2/g”
Make " particle diameter 45 μm filler, the specific surface area about 480m of routine into2/ g ", remaining is equal to embodiment 1.
Comparative example 2, by the aperture of the filter membrane 4 in embodiment 1 by 0.5 μm make into routine 0.45 μm;Remaining is equal to reality
Execute example 1.
Detect as tested 1 with the needle-based Solid-Phase Extraction filter of above-mentioned 2 comparative example gained, phosphatidylcholine
As described in Table 3 with the quantitative parameter of PHOSPHATIDYL ETHANOLAMINE.
Table 3
Finally, in addition it is also necessary to be only several specific embodiments of the present invention it is noted that listed above.Obviously, this
Bright it is not limited to above example, it is also possible to have many deformation.Those of ordinary skill in the art can be from present disclosure
The all deformation directly derived or associate, are all considered as protection scope of the present invention.
Claims (6)
1. needle-based Solid-Phase Extraction filter, is characterized in that: include housing (5), be respectively provided with in the both sides up and down of housing (5) with
The syringe nipple interface (6) of housing (5) intracavity inter-connection and pintle interface (7);
Upper sieve plate (1), solid phase extraction filler (2), lower sieve plate (3) and filter membrane is set gradually from top to bottom in housing (5) inner chamber
(4);
The sieve aperture that aperture is 95~105 μm it is respectively equipped with on described upper sieve plate (1) and lower sieve plate (3).
Needle-based Solid-Phase Extraction filter the most according to claim 1, is characterized in that: described solid phase extraction filler (2)
Particle diameter is 110~130 μm.
Needle-based Solid-Phase Extraction filter the most according to claim 2, is characterized in that: described solid phase extraction filler (2) for-
At least one in C18 ,-C8, silica gel, aluminium oxide, Florisil, kieselguhr, activated carbon, Diol (glycol-based).
4., according to the arbitrary described needle-based Solid-Phase Extraction filter of claims 1 to 3, it is characterized in that: described filter membrane (4) is Buddhist nun
Dragon.
Needle-based Solid-Phase Extraction filter the most according to claim 4, is characterized in that: the pore size filter of described filter membrane (4)
It it is 0.5 μm.
Needle-based Solid-Phase Extraction filter the most according to claim 1, is characterized in that: syringe nipple interface (6) and pin
Bolt mouth (7) is standard straight socket or standard thread mouth.
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Cited By (5)
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CN108469466A (en) * | 2018-03-23 | 2018-08-31 | 中国检验检疫科学研究院 | A kind of on-line extraction purification needle point electrospray ionization device and sample fast determining method |
CN109916700A (en) * | 2019-03-26 | 2019-06-21 | 华东理工大学 | A kind of small-sized Solid Phase Extraction and solid phase spectra detect aggregate device |
CN110691635A (en) * | 2017-11-17 | 2020-01-14 | 株式会社Lg化学 | Micro device for solid phase extraction |
CN111405933A (en) * | 2018-08-21 | 2020-07-10 | 株式会社Lg化学 | Solid phase extraction method using micro device |
CN111474249A (en) * | 2020-03-11 | 2020-07-31 | 台州学院 | Method for detecting organic alcohol amine compounds in environmental water sample |
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Cited By (9)
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CN110691635A (en) * | 2017-11-17 | 2020-01-14 | 株式会社Lg化学 | Micro device for solid phase extraction |
US11065557B2 (en) | 2017-11-17 | 2021-07-20 | Lg Chem, Ltd. | Micro device for solid phase extraction |
CN110691635B (en) * | 2017-11-17 | 2021-08-17 | 株式会社Lg化学 | Micro device for solid phase extraction |
CN108469466A (en) * | 2018-03-23 | 2018-08-31 | 中国检验检疫科学研究院 | A kind of on-line extraction purification needle point electrospray ionization device and sample fast determining method |
CN111405933A (en) * | 2018-08-21 | 2020-07-10 | 株式会社Lg化学 | Solid phase extraction method using micro device |
CN111405933B (en) * | 2018-08-21 | 2021-12-07 | 株式会社Lg化学 | Solid phase extraction method using micro device |
US11260319B2 (en) | 2018-08-21 | 2022-03-01 | Lg Chem, Ltd. | Solid phase extraction method using micro device |
CN109916700A (en) * | 2019-03-26 | 2019-06-21 | 华东理工大学 | A kind of small-sized Solid Phase Extraction and solid phase spectra detect aggregate device |
CN111474249A (en) * | 2020-03-11 | 2020-07-31 | 台州学院 | Method for detecting organic alcohol amine compounds in environmental water sample |
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