CN103645264B - Pretreatment method of blood sample - Google Patents
Pretreatment method of blood sample Download PDFInfo
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- CN103645264B CN103645264B CN201310687036.7A CN201310687036A CN103645264B CN 103645264 B CN103645264 B CN 103645264B CN 201310687036 A CN201310687036 A CN 201310687036A CN 103645264 B CN103645264 B CN 103645264B
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Abstract
A kind of pretreatment method of blood sample, 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, and the foreign protein in blood plasma is separated and forms floccus, 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; Extract the high-efficiency liquid chromatographic-grade acidic aqueous solution with desmosine ion.Desmosine is attached in floccus, and follow-up floccus adds in high-efficiency liquid chromatographic-grade acidic aqueous solution, and desmosine dissolves, and extracts the high-efficiency liquid chromatographic-grade acidic aqueous solution with desmosine ion and detects for HPLC.The pretreatment method of blood sample of the technical program, without the need to steps such as the extraction of prior art, nitrogen dry up, simplifies blood sample pre-treatment step, saves pretreatment time, also do not need operating personnel's Real-Time Monitoring, reduce the dependence to people.
Description
Technical field
The present invention relates to field of medical technology, particularly a kind of pretreatment method of blood sample.
Background technology
Carry out pre-service to blood sample to refer to extract determinand from blood sample.Follow-up by the content by detecting this determinand, one-tenth gradation parameter, reach the object of disease examination, health examination.
Such as, in the prior art, elastic fibrous tissue's degraded that injury of lungs patient is rich in due to lung generates specific 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, clinical examination to injury of lungs and research can be realized according to DES concentration.But HPLC has comparatively harsh requirement to the pre-service before blood sample loading.
Due to complicated component in blood plasma, wherein DES content is relatively low, and traditional preprocess method is comparatively complicated.Traditional pretreatment method of blood sample comprises: in test tube, add solution A and blood plasma, and solution A is for separating of the foreign protein in blood plasma; Vortex oscillation, makes the solution A in test tube and blood plasma fully mix, and the foreign protein in blood plasma forms floccus, and desmosine is free in solution with ionic state; Centrifugal, test tube is placed in hydro-extractor and carries out centrifugal treating, floccus is separated with solution, and is deposited on bottom test tube; Extraction, from the upper solution test tube, the solution of extraction containing desmosine is in another test tube; Nitrogen dries up, and use nitrogen brushes the solution in another test tube, and dry up completely to liquid, the dried powder in test tube comprises desmosine composition, and whole nitrogen dries up process probably needs 7 to 8 hours; Redissolve, in the test tube that the dried powder dried up is housed, add B solution, dried powder dissolves again, and desmosine is wherein free in B solution with ionic condition; Filter, remove other granulometric impurities redissolved in solution, obtain the solution with desmosine.The solution with desmosine ion detects for HPLC.
Analyze the preprocess method of traditional blood sample, whole process is too loaded down with trivial details, and whole blood sample pretreatment time is always consuming time reaches tens hours.In nitrogen gas blow dry step, need to stop nitrogen brushing in time after drying up, otherwise dried powder may be blown away.And nitrogen gas blow dry step spended time is oversize, this to the technical requirement of operating personnel and dependence high especially, need operating personnel's Real-Time Monitoring.In addition, needed for traditional pretreatment method of blood sample, plasma sample is more, generally at least needs 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 pretreatment method of blood sample is too loaded down with trivial details.
For solving the problem, the invention provides a kind of pretreatment method of blood sample, described pretreatment method of blood sample 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, and the foreign protein in blood plasma is separated and forms floccus, 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;
Extract the high-efficiency liquid chromatographic-grade acidic aqueous solution with desmosine ion.
Alternatively, described high-efficiency liquid chromatographic-grade acidic aqueous solution is formic acid solution or trifluoroacetic acid solution, and the volume concentration range of described formic acid solution, trifluoroacetic acid solution is 0.5 ‰ ~ 1 ‰.
Alternatively, before the described floccus of extraction, vortex oscillation is carried out to the test tube that acetonitrile solvent and blood plasma are housed.
Alternatively, the method for described extraction floccus is: carry out centrifugal filtration process, and obtain the floccus dried, at described centrifugal filtration process, the range of speeds of hydro-extractor is 4000 ~ 5000rpm.
Alternatively, add high-efficiency liquid chromatographic-grade acidic acid solution in described floccus after, leave standstill 2 ~ 3min.
Alternatively, after leaving standstill, vortex oscillation is carried out to the test tube that high-efficiency liquid chromatographic-grade acidic aqueous solution is housed.
Alternatively, the time range of described vortex oscillation is 20 ~ 30s.
Alternatively, extracting the method with the high-efficiency liquid chromatographic-grade acidic aqueous solution of desmosine ion is: carry out centrifugal filtration process, obtain the high-efficiency liquid chromatographic-grade acidic aqueous solution with desmosine ion, at described centrifugal filtration process, the rotating speed of hydro-extractor is 10000rpm.
Alternatively, described centrifugal filtration time range is 2 ~ 3min.
Alternatively, in described pretreatment method of blood sample, the test tube of use comprises the first pipe, separable the second pipe be set in the first pipe,
Described second pipe has the first opening and the second opening, and described 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;
Filter membrane is provided with in described second pipe, described filter membrane surface is relative with described first opening, described filter membrane edge and the second inside pipe wall are fitted, and the second tube portion between described filter membrane and the first opening is used 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 second pipe, the solution in the second pipe flows into one first pipe, and floccus is deposited on filter membrane surface;
After extraction floccus, be separated the first pipe and manage with second, the second pipe box is loaded in another first pipe;
Extract the high-efficiency liquid chromatographic-grade acidic aqueous solution process with desmosine ion, 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, and desmosine can be made to 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, extracts the high-efficiency liquid chromatographic-grade acidic aqueous solution with desmosine ion afterwards and detects for HPLC.The pretreatment method of blood sample of the technical program, without the need to steps such as extraction of the prior art, nitrogen dry up, simplifies blood sample pre-treatment step, saves blood sample pretreatment time, does not also need operating personnel's Real-Time Monitoring, reduce the dependence to people.
Accompanying drawing explanation
Fig. 1 is the cross-sectional view of the test tube that the blood sample preprocessing process of the specific embodiment of the invention uses.
Embodiment
The invention provides a kind of pretreatment method of blood sample, this preprocess method is simple to operate, and step is few, saves time.
For enabling above-mentioned purpose of the present invention, feature and advantage more become apparent, and are described in detail specific embodiments of the invention below in conjunction with accompanying drawing.
The first step, get a clean tube, acetonitrile solvent and blood plasma is added in test tube, the volume ratio of blood plasma and acetonitrile is more than or equal to 1:3, under the effect of acetonitrile solvent, foreign protein in blood plasma is separated and forms floccus, and 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, needed for the present embodiment, Plasma volumes is very little.
In the present embodiment, the volume ratio of blood plasma and acetonitrile solvent is 1:3, can ensure that the desmosine in blood plasma is substantially all adsorbed in floccus like this, avoids there is desmosine in mixed solution and remains, and affect testing result.In a particular embodiment, the volume ratio of blood plasma and acetonitrile solvent is not limited to 1:3, and it is also feasible for being greater than 1:3.
In the present embodiment, can follow and first add the less liquid of mass density, after add the principle of the larger liquid of mass density, 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, blood plasma is added.The blood plasma that mass density is larger can spread fast in acetonitrile solvent, ensures that blood plasma fully mixes with acetonitrile solvent.
When blood plasma mixes with acetonitrile solvent, under the effect of acetonitrile solvent, the cohesion of foreign protein in blood plasma forms floccus, and the desmosine simultaneously in blood plasma is ionic state, and is attached in floccus along with the cohesion of foreign protein.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 a particular embodiment, except the high-efficiency liquid chromatographic-grade acetonitrile solvent of the present embodiment, the high-purity or ultrapure acetonitrile solvent that other are feasible also can be used.
Second step, vortex oscillation.Test tube is placed in turbula shaker, clogs the opening of test tube, carry out vortex oscillation process, vortex oscillation makes blood plasma and acetonitrile solvent fully mix, and promotes the formation of floccus.In vortex oscillation process, can stir simultaneously, the relatively large floccus in test tube is broken up.Due to desmosine except attachment is floccus surface, also may be wrapped in floccus, break up floccus and desmosine can be made fully to be exposed to floccus surface, like this in the solution of subsequent process, desmosine in floccus can fully be dissolved, accurate to ensure the desmosine content recorded.
In a particular embodiment, the time range of vortex oscillation 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 oscillation is 30s.
3rd step, centrifugal filtration.Carry out centrifugal filtration process.In centrifugal filtration process, solution is separated with floccus, and solution suffers centrifugal filtration in other containers, and the liquid in floccus is dried substantially, obtains dry floccus, the floccus of drying is put into another test tube.In the present embodiment, the filter membrane 14 that filter process uses 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 μm, can filter out the most indissoluble granulometric impurities in solution.
In a particular embodiment, centrifugal filtration process uses low temperature ultracentrifuge, and the range of speeds of hydro-extractor is 4000 ~ 5000rpm, and described cryogenic temperature is 4 DEG C, and whole centrifugal process continues 2 ~ 3min.If the rotating speed of hydro-extractor is lower than 4000rpm, be unfavorable for drying floccus, acetonitrile may be had remain in floccus.If the rotating speed of hydro-extractor is higher than 5000rpm, floccus can be made in the compacting of filter membrane surface over-bonded, be unfavorable for that follow-up floccus is separated and dissolve again.
4th step, after carrying out centrifugal filtration, floccus is contained in another new test tube, high-efficiency liquid chromatographic-grade acidic aqueous solution is added in this new test tube, leave standstill, 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 the crosslinked formed by three allysine side chains and a lysine side-chain, adsorb with ionic forms on floccus surface, 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 a particular embodiment, described high-efficiency liquid chromatographic-grade acidic aqueous solution is formic acid solution or trifluoroacetic acid solution, and the volume concentration range 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 a particular embodiment, the volume of high-efficiency liquid chromatographic-grade acidic aqueous solution approximates Plasma volumes, can realize fully soaking into of floccus.
5th step, after leaving standstill, carry out vortex oscillation, the time range of vortex oscillation is 20 ~ 30s, fully mixes to make floccus and high-efficiency liquid chromatographic-grade acidic aqueous solution.When vortex oscillation, can stir, break up floccus, be beneficial to the desmosine release in floccus.In the present embodiment, the time of vortex oscillation is 30s.
6th step, centrifugal filtration.In centrifugal filtration process, the high-efficiency liquid chromatographic-grade acidic aqueous solution being dissolved with desmosine enters in next test tube pipe by filter membrane fast, other impurity in solution, particulate then remain on filter membrane, and the high-efficiency liquid chromatographic-grade acidic aqueous solution being dissolved with desmosine is required solution.In the present embodiment, the range of speeds of hydro-extractor is 10000rpm, to ensure that solution is centrifuged in next test tube as far as possible completely.In a particular embodiment, centrifugal residing environment temperature is 4 DEG C, and centrifugation time scope is 2 ~ 3min.In the present embodiment, centrifugation time is 2min.
Through above blood sample preprocessing process, obtain the high-efficiency liquid chromatographic-grade acidic aqueous solution with desmosine ion, this high-efficiency liquid chromatographic-grade acidic aqueous solution can be used for follow-up high performance liquid chromatography and detects.
In pretreatment method of blood sample of the present invention, existing test tube can be used.In a particular embodiment, except the existing test tube of use, also can use a kind of new test tube, will this new test tube application in the present invention be elaborated below.
With reference to Fig. 1, the test tube of use comprises the first pipe 101 and separable the second pipe 102 be set in the first pipe 101.
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, second opening 122 by the first opening 121 and flows out for liquid.Described first opening 121 has the extension 123 that can lie across at the first pipe 101 edge of opening, and when the first pipe 101 and the second pipe 102 are set with, extension 123 frame is on the edge of opening of the first pipe 101.
With reference to Fig. 1, filter membrane 104 is provided with in described second pipe 102, described filter membrane 104 is near described second opening 122, described filter membrane 104 surface is relative with described first opening, described filter membrane edge and the second inside pipe wall are fitted, the edge surface of such as filter membrane 104 and the second inside pipe wall use glue to bond, and avoid 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 μm.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 be revealed quickly through filter membrane 104, 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 used 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 slow transits through filter membrane 104, the liquid flowed 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 pretreatment method of blood sample 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 quickly through filter membrane 104, enter in the first pipe 101, filter membrane 104 plays a role in filtering in this process.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 surface, and is fitted in filter membrane 104 surface.In the 6th step of pretreatment method of blood sample 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 stays filter membrane 104 surface.The high-efficiency liquid chromatographic-grade acidic aqueous solution with desmosine, quickly through filter membrane 104, enters in another clean first test tube 101, and as being used for the required filtrate of efficient liquid phase chromatographic analysis.
When extracting floccus using traditional test tube and there is the high-efficiency liquid chromatographic-grade acidic aqueous solution of desmosine, special filter centrifugal need be used, and need the solution of filtration 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 filtered out is transferred to the first pipe from the second pipe, simplifies operation.
In a particular 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 wall is fixedly connected with support 103 at the edge of support 103, support 103 in the form of sheets, fit by the surface of support 103 and the intimate surface of filter membrane 104.Support 103, for carrying filter membrane 104, avoids filter membrane 104 to break in the vortex oscillation process, high speed centrifugation process of pretreatment method of blood sample of the present invention.In a particular embodiment, support 103 flows out in cellular and in permission the second pipe 102 liquid.
In a particular embodiment, with reference to Fig. 1, the second inside pipe wall between described 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 the described floccus of extraction, when vortex oscillation is carried out to the test tube that acetonitrile solvent and blood plasma are housed; Or before extraction has the high-efficiency liquid chromatographic-grade acidic aqueous solution of desmosine ion, when carrying out vortex oscillation to the test tube that floccus and high-efficiency liquid chromatographic-grade acidic aqueous solution are housed, cutting member 107 can cut floccus, realize the object of breaing up floccus.With the vortex oscillation process using traditional test tube, the test tube of the present embodiment more easily operates, simple and easy to do.
In a particular 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 distribution, and cutting member 107 can be movable in accommodation space 106.In vortex oscillation process, the diameter due to cutting member 107 is less than the second pipe 102 internal diameter, rocks in the liquid of cutting member 107 in the second pipe 102, rocks mesh lines cutting floccus in process, realizes the object of breaing up floccus.In other embodiments, also can not arrange accommodation space, 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 oscillation, mesh lines also can cut floccus.Therefore, use the test tube of the present embodiment, while vortex oscillation, floccus can be broken up.
In a particular 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, the one end open of tapered portion 108 is docked with the second opening 122 of the second pipe 102, the other end opening of tapered portion 108 is positioned at outside the second pipe 102, and the other end opening inside diameter of tapered portion 108 is less than the second opening 122 internal diameter.In the High Rotation Speed of centrifugal filtration process, thrown away at a high speed by the solution droplets of filter membrane 104, tapered portion 108 can stop that the drop of splashing flies to the first pipe 101 inwall, and drop slowly can enter the bottom of the first pipe 101 by the opening with less internal diameter of tapered portion 108 along the inwall of tapered portion 108.
In a particular embodiment, with reference to Fig. 1, described test tube also comprises lid 105, and lid 105 comprises the body 151 being covered on the second pipe first opening 121, the junction of handle 152, first pipe outer wall between handle 152 with body 151 be connected with body 151 is connected with lid 105.At vortex oscillation, 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.
Use the new test tube of the present embodiment, the filter membrane in the second pipe plays a role in filtering, and the cutting member in the second pipe can realize breaing up floccus effect.Compared with traditional test tube, new test tube has multiple function, can improve blood sample pre-service efficiency, saves pretreatment time.In addition, the second pipe can realize 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 use 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, and therefore protection scope of the present invention should be as the criterion with claim limited range.
Claims (10)
1. a pretreatment method of blood sample, 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, and the foreign protein in blood plasma is separated and forms floccus, 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;
Extract the high-efficiency liquid chromatographic-grade acidic aqueous solution with desmosine ion.
2. pretreatment method of blood sample as claimed in claim 1, it is characterized in that, described high-efficiency liquid chromatographic-grade acidic aqueous solution is formic acid solution or trifluoroacetic acid solution, and the volume concentration range of described formic acid solution, trifluoroacetic acid solution is 0.5 ‰ ~ 1 ‰.
3. pretreatment method of blood sample as claimed in claim 1, is characterized in that, before the described floccus of extraction, carries out vortex oscillation to the test tube that acetonitrile solvent and blood plasma are housed.
4. pretreatment method of blood sample as claimed in claim 1, it is characterized in that, the method for described extraction floccus is: carry out centrifugal filtration process, and obtain the floccus dried, at described centrifugal filtration process, the range of speeds of hydro-extractor is 4000 ~ 5000rpm.
5. pretreatment method of blood sample as claimed in claim 1, is characterized in that, after adding high-efficiency liquid chromatographic-grade acidic acid solution, leaves standstill 2 ~ 3min in described floccus.
6. pretreatment method of blood sample as claimed in claim 5, is characterized in that, after leaving standstill, carries out vortex oscillation to the test tube that high-efficiency liquid chromatographic-grade acidic aqueous solution is housed.
7. the pretreatment method of blood sample as described in claim 3 or 6, is characterized in that, the time range of described vortex oscillation is 20 ~ 30s.
8. pretreatment method of blood sample as claimed in claim 1, it is characterized in that, extracting the method with the high-efficiency liquid chromatographic-grade acidic aqueous solution of desmosine ion is: carry out centrifugal filtration process, obtain the high-efficiency liquid chromatographic-grade acidic aqueous solution with desmosine ion, at described centrifugal filtration process, the rotating speed of hydro-extractor is 10000rpm.
9. the pretreatment method of blood sample as described in claim 4 or 8, is characterized in that, described centrifugal filtration time range is 2 ~ 3min.
10. pretreatment method of blood sample as claimed in claim 1, is characterized in that, in described pretreatment method of blood sample, the test tube of use comprises the first pipe, separable the second pipe be set in the first pipe,
Described second pipe has the first opening and the second opening, and described 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;
Filter membrane is provided with in described second pipe, described filter membrane surface is relative with described first opening, described filter membrane edge and the second inside pipe wall are fitted, and the second tube portion between described filter membrane and the first opening is used 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 second pipe, the solution in the second pipe flows into the first pipe, and floccus is deposited on filter membrane surface;
After extraction floccus, be separated the first pipe and manage with second, the second pipe box is loaded in another first pipe;
Extract the high-efficiency liquid chromatographic-grade acidic aqueous solution process with desmosine ion, 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 |
|---|
| Determination of free desmosine in human plasma and its application in two experimental medicine studies;Jens Lamerz等;《Analytical Biochemistry》;20130208;第436卷;127-136 * |
| Quantitation of desmosine and isodesmosine in urine, plasma, and sputum by LC–MS/MS as biomarkers for elastin degradation;Shuren Ma等;《Journal of Chromatography B》;20110513;第879卷;1893-1898 * |
| RP-HPLC法测定大鼠血浆中姜黄素含量;顾吉晋等;《成都医学院学报》;20091231;第4卷(第4期);文章第2.4.1节 * |
| 高效液相色谱法检测弹性蛋白中锁链素;孙炎等;《色谱》;19970531;第15卷(第3期);235-236 * |
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