CN103645264A - Pretreatment method of blood sample - Google Patents
Pretreatment method of blood sample Download PDFInfo
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- CN103645264A CN103645264A CN201310687036.7A CN201310687036A CN103645264A CN 103645264 A CN103645264 A CN 103645264A CN 201310687036 A CN201310687036 A CN 201310687036A CN 103645264 A CN103645264 A CN 103645264A
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Abstract
The invention discloses a pretreatment method of a blood sample. The method comprises the following steps: adding an acetonitrile solvent and plasma to a test tube, wherein the volume ratio of the plasma to the acetonitrile solvent is greater than or equal to 1:3; separating foreign proteins in the plasma to form a floccule; attaching desmosine in the floccule; extracting the floccule; adding a high-performance liquid chromatography acidic water solution to the floccule, wherein the desmosine in the floccule drifts away in the high-performance liquid chromatography acidic water solution in an ionic form; extracting the high-performance liquid chromatography acidic water solution with desmosine ions; attaching the desmosine in the floccule; adding subsequent floccule to the high-performance liquid chromatography acidic water solution; dissolving the desmosine and extracting the high-performance liquid chromatography acidic water solution with the desmosine ions for high-performance liquid chromatography (HPLC) detection. The pretreatment method of the blood sample in the technical scheme does not need the steps of extraction, nitrogen drying and the like in the prior art, the pretreatment step of the blood sample is simplified, the pretreatment time is saved, an operator is not needed to monitor in real time, and the dependency on people is reduced.
Description
Technical field
The present invention relates to field of medical technology, particularly a kind of blood sample preprocess method.
Background technology
Blood sample is carried out to pre-service to be referred to and from blood sample, extracts determinand.Follow-up by by detecting content, the one-tenth gradation parameter of this determinand, reach the object of disease examination, health examination.
For example, in the prior art, elastic fibrous tissue's degraded that injury of lungs patient is rich in due to lung generates specificity product desmosine (desmosine, DES), causes the rising of desmosine concentration in blood plasma.Therefore, use high performance liquid chromatography (High Performance Liquid Chromatography, HPLC) to detect DES concentration in blood plasma, according to DES concentration, can realize the clinical examination of injury of lungs and research.But the pre-service of HPLC before to blood sample loading has comparatively harsh requirement.
Due to complicated component in blood plasma, wherein DES content is relatively low, and traditional preprocess method is comparatively complicated.Traditional blood sample preprocess method comprises: in test tube, add A solution and blood plasma, A solution is for separating of the foreign protein in blood plasma; Vortex vibration, fully mixes A solution and blood plasma in test tube, and the foreign protein in blood plasma forms floccus, and desmosine is free in solution with ionic state; Centrifugal, test tube is placed in to hydro-extractor and carries out centrifugal treating, floccus is separated with solution, and is deposited on test tube bottom; Extraction, the solution that in the upper solution from test tube, extraction contains desmosine is to another test tube; Nitrogen dries up, and uses nitrogen to brush the solution in another test tube, dries up completely to liquid, and the dried powder in test tube comprises desmosine composition, and whole nitrogen dries up process probably needs 7 to 8 hours; Redissolve, to being equipped with in the test tube of the dried powder drying up, add B solution, dried powder dissolves again, and desmosine is wherein free in B solution with ionic condition; Filter, remove other granulometric impurities that redissolve in solution, obtain having the solution of desmosine.The solution with desmosine ion detects for HPLC.
Preprocess method to traditional blood sample is analyzed, and whole process is too loaded down with trivial details, and whole blood sample pretreatment time is always consuming time reaches tens hours.Nitrogen dries up in step, need to stop in time nitrogen and brush, otherwise dried powder may be blown away after drying up.And nitrogen to dry up step spended time oversize, this is high especially to operating personnel's technical requirement and dependence, needs operating personnel's Real-Time Monitoring.In addition, traditional required plasma sample of blood sample preprocess method is more, generally at least need 0.5～1mL, and utilization factor is also low.
Summary of the invention
The problem that the present invention solves is that the process of traditional blood sample preprocess method is too loaded down with trivial details.
For addressing the above problem, the invention provides a kind of blood sample preprocess method, described blood sample preprocess method comprises:
In test tube, add acetonitrile solvent and blood plasma, the volume ratio of blood plasma and acetonitrile solvent is more than or equal to 1:3, the separated floccus that forms of foreign protein in blood plasma, and desmosine is attached in floccus;
Extract floccus;
In described floccus, add high-efficiency liquid chromatographic-grade acidic aqueous solution, the desmosine in floccus is free in described high-efficiency liquid chromatographic-grade acidic aqueous solution with ionic forms;
Extraction has the high-efficiency liquid chromatographic-grade acidic aqueous solution of desmosine ion.
Alternatively, described high-efficiency liquid chromatographic-grade acidic aqueous solution is formic acid solution or trifluoroacetic acid solution, and the volumetric concentration scope of described formic acid solution, trifluoroacetic acid solution is 0.5 ‰～1 ‰.
Alternatively, before extracting described floccus, to being housed, the test tube of acetonitrile solvent and blood plasma carries out vortex vibration.
Alternatively, the method for described extraction floccus is: carry out centrifugal filtration processing, obtain the floccus of drying, in described centrifugal filtration process, the range of speeds of hydro-extractor is 4000～5000rpm.
Alternatively, in described floccus, add after high-efficiency liquid chromatographic-grade acidic acid solution standing 2～3min.
Alternatively, standing after, to the test tube of high-efficiency liquid chromatographic-grade acidic aqueous solution is housed, carry out vortex vibration.
Alternatively, the time range of described vortex vibration is 20～30s.
Alternatively, the method that extraction has the high-efficiency liquid chromatographic-grade acidic aqueous solution of desmosine ion is: carry out centrifugal filtration processing, obtain having the high-efficiency liquid chromatographic-grade acidic aqueous solution of desmosine ion, in described centrifugal filtration process, the rotating speed of hydro-extractor is 10000rpm.
Alternatively, described centrifugal filtration time range is 2～3min.
Alternatively, in described blood sample preprocess method, the test tube of use comprises the first pipe, the separable pipe of second in the first pipe that is set in,
Described the second pipe has the first opening and the second opening, and described the first opening has the extension that can lie across at the first tube opening edge, and acetonitrile solvent, blood plasma and high-efficiency liquid chromatographic-grade acidic aqueous solution enter in the second pipe by the first opening;
In described the second pipe, be provided with filter membrane, described filter membrane surface is relative with described the first opening, described filter membrane edge and the laminating of the second inside pipe wall, the second tube portion between described filter membrane and the first opening is for splendid attire acetonitrile solvent, blood plasma and high-efficiency liquid chromatographic-grade acidic aqueous solution;
Extract floccus process, use the solution in membrane filtration the second pipe, the solution in the second pipe flows into one first pipe, and floccus is deposited on filter membrane surface;
After extracting floccus, separated the first pipe and the second pipe, be loaded on the second pipe box in another first pipe;
Extraction has the high-efficiency liquid chromatographic-grade acidic aqueous solution process of desmosine ion, and the high-efficiency liquid chromatographic-grade acidic aqueous solution with desmosine ion flows into another the first pipe.
Compared with prior art, technical scheme of the present invention has the following advantages:
The volume ratio of blood plasma and acetonitrile solvent is more than or equal to 1:3, can make desmosine be attached in floccus.Because desmosine is attached in floccus, follow-up floccus adds in high-efficiency liquid chromatographic-grade acidic aqueous solution, and desmosine discharges and dissolves from floccus, and the high-efficiency liquid chromatographic-grade acidic aqueous solution that extraction afterwards has desmosine ion detects for HPLC.The blood sample preprocess method of the technical program, without steps such as extraction of the prior art, nitrogen dry up, has been simplified blood sample pre-treatment step, saves blood sample pretreatment time, does not also need operating personnel's Real-Time Monitoring, reduces the dependence to people.
Accompanying drawing explanation
Fig. 1 is the cross-sectional view of the test tube that uses of the blood sample preprocessing process of the specific embodiment of the invention.
Embodiment
The invention provides a kind of blood sample preprocess method, this preprocess method is simple to operate, and step is few, saves time.
For above-mentioned purpose of the present invention, feature and advantage can more be become apparent, below in conjunction with accompanying drawing, specific embodiments of the invention are described in detail.
The first step, get a clean tube, in test tube, add acetonitrile solvent and blood plasma, the volume ratio of blood plasma and acetonitrile is more than or equal to 1:3, under the effect of acetonitrile solvent, the separated floccus that forms of foreign protein in blood plasma, the desmosine in blood plasma is attached in floccus with ionic state, and floccus is arranged in the solution of test tube.In the present embodiment, the volume range of blood plasma is 75 μ L～100 μ L, and compared with prior art, the required blood plasma volume of the present embodiment is very little.
In the present embodiment, the volume ratio of blood plasma and acetonitrile solvent is 1:3, can guarantee that like this desmosine in blood plasma is substantially all adsorbed in floccus, avoids existing desmosine residual in mixed solution, and affects testing result.In specific embodiment, the volume ratio of blood plasma and acetonitrile solvent is not limited to 1:3, and it is also feasible being greater than 1:3.
In the present embodiment, can follow the liquid that first adds mass density less, after add the principle of the liquid that mass density is larger, first in test tube, add acetonitrile solvent, the mass density of acetonitrile solvent is less than the mass density of blood plasma; Then in acetonitrile solvent, add blood plasma.The blood plasma that mass density is larger can spread fast in acetonitrile solvent, guarantees that blood plasma fully mixes with acetonitrile solvent.
When blood plasma mixes with acetonitrile solvent, under the effect of acetonitrile solvent, the foreign protein cohesion in blood plasma forms floccus, and the desmosine in blood plasma is ionic state simultaneously, and along with the cohesion of foreign protein is attached in floccus.In the present embodiment, acetonitrile solvent is high-efficiency liquid chromatographic-grade acetonitrile solvent, and so-called high-efficiency liquid chromatographic-grade refers to that the purity of acetonitrile is very high, the reagent purity that can use in high performance liquid chromatography.In specific embodiment, except the high-efficiency liquid chromatographic-grade acetonitrile solvent of the present embodiment, also can use other feasible high-purity or ultrapure acetonitrile solvents.
Second step, vortex vibration.Test tube is placed in to vortex oscillator, clogs the opening of test tube, carry out vortex oscillation treatment, vortex vibration fully mixes blood plasma and acetonitrile solvent, promotes the formation of floccus.In vortex oscillatory process, can stir simultaneously, the relatively large floccus in test tube is broken up.Because desmosine is floccus surface except adhering to, also may be wrapped in floccus, break up floccus and can make desmosine fully be exposed to floccus surface, like this in the solution of subsequent process, desmosine in floccus can fully be dissolved, accurate with the desmosine content that guarantees to record.
In specific embodiment, the time range of vortex vibration is 20～30s, can realize blood plasma and acetonitrile solvent fully mixes, and breaks up relatively large floccus.In the present embodiment, the time of vortex vibration is 30s.
The 3rd step, centrifugal filtration.Carry out centrifugal filtration processing.In centrifugal filtration process, solution is separated with floccus, and solution suffers centrifugal filtration to other containers, and the liquid in floccus is dried substantially, obtains dry floccus, and dry floccus is put into another test tube.In the present embodiment, the filter membrane 14 that filter process is used is organic phase filter membrane, allows organic solvent to pass through.The filter opening aperture of filter membrane 14 is not more than 0.5 μ m, can filter out the most indissoluble granulometric impurities in solution.
In specific embodiment, centrifugal filtration process is used low temperature ultracentrifuge, and the range of speeds of hydro-extractor is 4000～5000rpm, and described cryogenic temperature is 4 ℃, and whole centrifugal process continues 2～3min.If the rotating speed of hydro-extractor, lower than 4000rpm, is unfavorable for drying floccus, in floccus, acetonitrile may be had residual.If the rotating speed of hydro-extractor higher than 5000rpm, can make floccus in the over-bonded compacting of filter membrane surface, be unfavorable for that follow-up floccus separation dissolves again.
The 4th step, after carrying out centrifugal filtration, floccus is contained in another new test tube, in this new test tube, add high-efficiency liquid chromatographic-grade acidic aqueous solution, standing, time of repose scope 2～3min, fully soaks in high-efficiency liquid chromatographic-grade acidic aqueous solution to realize floccus.In the present embodiment, time of repose is 2min.
Desmosine is by three allysine side chains and a crosslinked that lysine side-chain forms, on floccus surface, with ionic forms, adsorb, under the effect of high-efficiency liquid chromatographic-grade acidic aqueous solution, the desmosine on floccus surface easily departs from floccus surface, and desmosine is free in high-efficiency liquid chromatographic-grade acidic aqueous solution with ionic forms.
In specific embodiment, described high-efficiency liquid chromatographic-grade acidic aqueous solution is formic acid solution or trifluoroacetic acid solution, and the volumetric concentration scope of high-efficiency liquid chromatographic-grade acidic aqueous solution was 0.5 ‰～1 ‰ (comprising end points).In the present embodiment, described high-efficiency liquid chromatographic-grade acidic aqueous solution is trifluoroacetic acid solution, and the volumetric concentration of trifluoroacetic acid solution is 1 ‰.In follow-up efficient liquid phase chromatographic analysis process, high-efficiency liquid chromatographic-grade acidic aqueous solution is as mobile phase.
In specific embodiment, the volume of high-efficiency liquid chromatographic-grade acidic aqueous solution approximates blood plasma volume, can realize fully soaking into of floccus.
The 5th step, after standing, carries out vortex vibration, and the time range of vortex vibration is 20～30s, so that floccus and high-efficiency liquid chromatographic-grade acidic aqueous solution fully mix.When vortex vibrates, can stir, break up floccus, the desmosine being beneficial in floccus discharges.In the present embodiment, the time of vortex vibration is 30s.
The 6th step, centrifugal filtration.In centrifugal filtration process, the high-efficiency liquid chromatographic-grade acidic aqueous solution that is dissolved with desmosine enters in next test tube pipe fast by filter membrane, other impurity, particulate in solution remain on filter membrane, and the high-efficiency liquid chromatographic-grade acidic aqueous solution that is dissolved with desmosine is required solution.In the present embodiment, the range of speeds of hydro-extractor is 10000rpm, to guarantee that solution is centrifuged in next test tube as far as possible completely.In specific embodiment, centrifugal environment temperature of living in is 4 ℃, and centrifugation time scope is 2～3min.In the present embodiment, centrifugation time is 2min.
Through above blood sample preprocessing process, obtain having the high-efficiency liquid chromatographic-grade acidic aqueous solution of desmosine ion, this high-efficiency liquid chromatographic-grade acidic aqueous solution can be used for follow-up high performance liquid chromatography and detects.
In blood sample preprocess method of the present invention, can use existing test tube.In specific embodiment, except using existing test tube, also can use a kind of new test tube, will elaborate this new test tube application in the present invention below.
With reference to Fig. 1, the test tube of use comprises the first pipe 101 and the separable pipe of second in the first pipe 101 102 that is set in.
The second pipe 102 has the first opening 121 and the second opening 122.Liquid, as blood plasma, acetonitrile solvent, high-efficiency liquid chromatographic-grade acidic aqueous solution, enters the second pipe 102, the second openings 122 by the first opening 121 and flows out for liquid.Described the first opening 121 has the extension 123 that can lie across at the first pipe 101 edge of opening, and in the first pipe 101 and second 102 whens suit of pipe, 123 of extensions are on the edge of opening of the first pipe 101.
With reference to Fig. 1, in described the second pipe 102, be provided with filter membrane 104, described filter membrane 104 is near described the second opening 122, described filter membrane 104 surfaces are relative with described the first opening, described filter membrane edge and the laminating of the second inside pipe wall, it is bonding that for example the edge surface of filter membrane 104 and the second inside pipe wall are used glue, avoids the impurity in the second pipe 102 solution to pass through from the gap between filter membrane 104 and the inwall of the second pipe 102.
For the test tube of the present embodiment, the first, the filter opening aperture of filter membrane 104 is less, and pore diameter range is not more than 0.5 μ m.Like this, when blood plasma, acetonitrile solvent and high-efficiency liquid chromatographic-grade acidic aqueous solution enter the second pipe 102 by the first opening 121, liquid can not revealed by filter membrane 104 fast, liquid is extremely slow by the speed of filter membrane 104, and the second tube portion between filter membrane 104 and the first opening 121 is for splendid attire blood plasma, acetonitrile solvent and high-efficiency liquid chromatographic-grade acidic aqueous solution.And the second pipe 102 is set in the first pipe 101, even if liquid slowly passes through filter membrane 104, the liquid flowing down enters the first pipe 101, there will not be liquid to spill, and also can not affect testing result.
Second, in the 3rd step of blood sample preprocess method of the present invention, extract the centrifugal filtration process of floccus, the acetonitrile solvent in the second pipe 102 and the liquid component in blood plasma are fast by filter membrane 104, enter in the first pipe 101, in this process, filter membrane 104 plays a role in filtering.When high speed centrifugation, the acetonitrile solvent in floccus is dried substantially completely, obtains dry floccus, and dry floccus is deposited on filter membrane 104 surfaces, and is fitted in filter membrane 104 surfaces.In the 6th step of blood sample preprocess method of the present invention, extract the high-efficiency liquid chromatographic-grade acidic aqueous solution process with desmosine, filter membrane 104 plays a role in filtering, and the impurity in solution is stayed filter membrane 104 surfaces.There is the high-efficiency liquid chromatographic-grade acidic aqueous solution of desmosine fast by filter membrane 104, enter in another clean first test tube 101, and as the required filtrate for efficient liquid phase chromatographic analysis.
When using traditional test tube extract floccus and there is the high-efficiency liquid chromatographic-grade acidic aqueous solution of desmosine, need to use special filter centrifugal, and the solution of filtration need to be transferred in another test tube by a test tube.By comparison, use the new test tube of the present embodiment, in general hydro-extractor, just can realize centrifugal filtration, and the solution filtering out transfers to the first pipe from the second pipe, simplified operation.
In specific embodiment, with reference to Fig. 1, between filter membrane 104 and the second opening 122, be provided with porous support 103, the second inside pipe walls and be fixedly connected with support 103 at the edge of support 103, in the form of sheets, the surface of the surface of support 103 and filter membrane 104 fits tightly support 103.Support 103 is for carrying filter membrane 104, avoids filter membrane 104 to break in the vortex oscillatory process of blood sample preprocess method of the present invention, high speed centrifugation process.In specific embodiment, support 103 is cellular and allows the liquid in the second pipe 102 to flow out.
In specific embodiment, with reference to Fig. 1, the second inside pipe wall between described the first opening 121 and filter membrane 104 is provided with annular element 109, described annular element 109 is around the axis of the second pipe 102, space between described filter membrane 104 and annular element 109, as accommodation space 106, is provided with porous cutting member 107 in described accommodation space 106.Before extracting described floccus, while carrying out vortex vibration to the test tube of acetonitrile solvent and blood plasma is housed; Or have the high-efficiency liquid chromatographic-grade acidic aqueous solution of desmosine ion in extraction before, while carrying out vortex vibration to the test tube of floccus and high-efficiency liquid chromatographic-grade acidic aqueous solution is housed, cutting member 107 can cut floccus, realizes the object of breaing up floccus.With the vortex oscillatory process of using traditional test tube, the test tube of the present embodiment more easily operates, simple and easy to do.
In specific embodiment, in the form of sheets, porous cutting member 107 is positioned at the surface of filter membrane 104 to this porous cutting member 107, and the diameter of cutting member 107 is less than the internal diameter of the second pipe 102.The porous of cutting member 107 presents grid and distributes, and cutting member 107 can be movable in accommodation space 106.In vortex oscillatory process, because the diameter of cutting member 107 is less than the second pipe 102 internal diameters, in the liquid of cutting member 107 in the second pipe 102, rock, rock mesh lines cutting floccus in process, realize the object of breaing up floccus.In other embodiments, also accommodation space can be set, cutting member 107 is fixedly connected with the second inside pipe wall, and spaced apart a little between cutting member 107 and filter membrane 104, and when vortex vibrates, mesh lines also can cut floccus.Therefore, use the test tube of the present embodiment, can, in vortex vibration, break up floccus.
In specific embodiment, with reference to Fig. 1, test tube also comprises the tapered portion 108 of a hollow, truncated, the two ends of this tapered portion 108 have opening, one end opening of tapered portion 108 docks with the second opening 122 of the second pipe 102, the other end opening of tapered portion 108 is positioned at the second pipe 102 outsides, and the other end opening internal diameter of tapered portion 108 is less than the second opening 122 internal diameters.In the High Rotation Speed of centrifugal filtration process, solution droplets by filter membrane 104 is thrown away at a high speed, tapered portion 108 can stop that the drop splashing flies to the first pipe 101 inwalls, and drop can slowly enter the bottom of the first pipe 101 along the inwall of tapered portion 108 by the opening with less internal diameter of tapered portion 108.
In specific embodiment, with reference to Fig. 1, described test tube also comprises lid 105, lid 105 comprise be covered on the body 151 of second pipe the first opening 121, handle 152, the first pipe outer walls that are connected with body 151 are connected with lid 105 with the junction between body 151 at handle 152.In vortex vibration, centrifugal filtration process, lid 105 covers the first opening 121 of the second pipe 102, avoids the liquid in the second pipe 102 to splash away from the first opening 121.
The new test tube that uses the present embodiment, the filter membrane in the second pipe plays a role in filtering, and the cutting member in the second pipe can be realized the floccus effect of breaing up.Compare with traditional test tube, new test tube has a plurality of functions, can improve blood sample pre-service efficiency, saves pretreatment time.In addition, the second pipe can be realized separable suit with the first pipe, and this makes the Renewal process of the first pipe convenient especially, and the first pipe also can repeatedly be used in a preprocessing process.
Although the present invention discloses as above, the present invention is not defined in this.Any those skilled in the art, without departing from the spirit and scope of the present invention, all can make various changes or modifications, so protection scope of the present invention should be as the criterion with claim limited range.
Claims (10)
1. a blood sample preprocess method, is characterized in that, comprising:
In test tube, add acetonitrile solvent and blood plasma, the volume ratio of blood plasma and acetonitrile solvent is more than or equal to 1:3, the separated floccus that forms of foreign protein in blood plasma, and desmosine is attached in floccus;
Extract floccus;
In described floccus, add high-efficiency liquid chromatographic-grade acidic aqueous solution, the desmosine in floccus is free in described high-efficiency liquid chromatographic-grade acidic aqueous solution with ionic forms;
Extraction has the high-efficiency liquid chromatographic-grade acidic aqueous solution of desmosine ion.
2. blood sample preprocess method as claimed in claim 1, is characterized in that, described high-efficiency liquid chromatographic-grade acidic aqueous solution is formic acid solution or trifluoroacetic acid solution, and the volumetric concentration scope of described formic acid solution, trifluoroacetic acid solution is 0.5 ‰～1 ‰.
3. blood sample preprocess method as claimed in claim 1, is characterized in that, before extracting described floccus, to the test tube of acetonitrile solvent and blood plasma is housed, carries out vortex vibration.
4. blood sample preprocess method as claimed in claim 1, is characterized in that, the method for described extraction floccus is: carry out centrifugal filtration processing, obtain the floccus of drying, in described centrifugal filtration process, the range of speeds of hydro-extractor is 4000～5000rpm.
5. blood sample preprocess method as claimed in claim 1, is characterized in that, adds after high-efficiency liquid chromatographic-grade acidic acid solution standing 2～3min in described floccus.
6. blood sample preprocess method as claimed in claim 5, is characterized in that, standing after, to the test tube of high-efficiency liquid chromatographic-grade acidic aqueous solution is housed, carry out vortex vibration.
7. the blood sample preprocess method as described in claim 3 or 6, is characterized in that, the time range of described vortex vibration is 20～30s.
8. blood sample preprocess method as claimed in claim 1, it is characterized in that, the method that extraction has the high-efficiency liquid chromatographic-grade acidic aqueous solution of desmosine ion is: carry out centrifugal filtration processing, obtain having the high-efficiency liquid chromatographic-grade acidic aqueous solution of desmosine ion, in described centrifugal filtration process, the rotating speed of hydro-extractor is 10000rpm.
9. the blood sample preprocess method as described in claim 4 or 8, is characterized in that, described centrifugal filtration time range is 2～3min.
10. blood sample preprocess method as claimed in claim 1, is characterized in that, in described blood sample preprocess method, the test tube of use comprises the first pipe, the separable pipe of second in the first pipe that is set in,
Described the second pipe has the first opening and the second opening, and described the first opening has the extension that can lie across at the first tube opening edge, and acetonitrile solvent, blood plasma and high-efficiency liquid chromatographic-grade acidic aqueous solution enter in the second pipe by the first opening;
In described the second pipe, be provided with filter membrane, described filter membrane surface is relative with described the first opening, described filter membrane edge and the laminating of the second inside pipe wall, the second tube portion between described filter membrane and the first opening is for splendid attire acetonitrile solvent, blood plasma and high-efficiency liquid chromatographic-grade acidic aqueous solution;
Extract floccus process, use the solution in membrane filtration the second pipe, the solution in the second pipe flows into one first pipe, and floccus is deposited on filter membrane surface;
After extracting floccus, separated the first pipe and the second pipe, be loaded on the second pipe box in another first pipe;
Extraction has the high-efficiency liquid chromatographic-grade acidic aqueous solution process of desmosine ion, and the high-efficiency liquid chromatographic-grade acidic aqueous solution with desmosine ion flows into another the first pipe.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4083788A (en) * | 1975-11-19 | 1978-04-11 | Ferrara Louis T | Blood serum-isolation device |
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2013
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4083788A (en) * | 1975-11-19 | 1978-04-11 | Ferrara Louis T | Blood serum-isolation device |
Non-Patent Citations (4)
| Title |
|---|
| JENS LAMERZ等: "Determination of free desmosine in human plasma and its application in two experimental medicine studies", 《ANALYTICAL BIOCHEMISTRY》 * |
| SHUREN MA等: "Quantitation of desmosine and isodesmosine in urine, plasma, and sputum by LC–MS/MS as biomarkers for elastin degradation", 《JOURNAL OF CHROMATOGRAPHY B》 * |
| 孙炎等: "高效液相色谱法检测弹性蛋白中锁链素", 《色谱》 * |
| 顾吉晋等: "RP-HPLC法测定大鼠血浆中姜黄素含量", 《成都医学院学报》 * |
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