CN114324689A - Preparation method of blank serum - Google Patents
Preparation method of blank serum Download PDFInfo
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- CN114324689A CN114324689A CN202111654145.XA CN202111654145A CN114324689A CN 114324689 A CN114324689 A CN 114324689A CN 202111654145 A CN202111654145 A CN 202111654145A CN 114324689 A CN114324689 A CN 114324689A
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- serum
- quantitative ring
- pump
- purified
- switched
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- 210000002966 serum Anatomy 0.000 title claims abstract description 75
- 238000002360 preparation method Methods 0.000 title claims abstract description 8
- 239000007788 liquid Substances 0.000 claims abstract description 30
- 238000000746 purification Methods 0.000 claims abstract description 30
- 239000012086 standard solution Substances 0.000 claims abstract description 9
- 150000001875 compounds Chemical class 0.000 claims abstract description 6
- 238000000034 method Methods 0.000 claims description 12
- 239000002904 solvent Substances 0.000 claims description 9
- 239000002245 particle Substances 0.000 claims description 7
- 239000011347 resin Substances 0.000 claims description 7
- 229920005989 resin Polymers 0.000 claims description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- 210000004369 blood Anatomy 0.000 claims description 4
- 239000008280 blood Substances 0.000 claims description 4
- 239000003513 alkali Substances 0.000 claims description 3
- 239000000284 extract Substances 0.000 claims description 3
- 230000002572 peristaltic effect Effects 0.000 claims description 3
- 238000002791 soaking Methods 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 238000005086 pumping Methods 0.000 claims description 2
- 238000002347 injection Methods 0.000 description 7
- 239000007924 injection Substances 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 239000011159 matrix material Substances 0.000 description 2
- 229920006395 saturated elastomer Polymers 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- HSINOMROUCMIEA-FGVHQWLLSA-N (2s,4r)-4-[(3r,5s,6r,7r,8s,9s,10s,13r,14s,17r)-6-ethyl-3,7-dihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-17-yl]-2-methylpentanoic acid Chemical compound C([C@@]12C)C[C@@H](O)C[C@H]1[C@@H](CC)[C@@H](O)[C@@H]1[C@@H]2CC[C@]2(C)[C@@H]([C@H](C)C[C@H](C)C(O)=O)CC[C@H]21 HSINOMROUCMIEA-FGVHQWLLSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 238000010811 Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003613 bile acid Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000002858 neurotransmitter agent Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
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- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention provides a preparation method of blank serum, which comprises the following steps: allowing the serum to enter a purifying column to remove endogenous compounds in the serum; the first multi-channel direction selection valve is switched, and purified serum is discharged from the purification column and enters a first quantitative ring; the first multi-channel direction selection valve is switched with the multi-way valve, the first pump pushes purified serum in the first quantitative ring, the purified serum enters the second quantitative ring, and two ends of the second quantitative ring are respectively connected with ports of the multi-way valve; switching the multi-way valve, wherein the carrier liquid pushes the purified serum in the second quantitative ring to enter an analyzer, and the analyzer gives parameters of the purified serum; calculating the deviation between the parameters and the parameters of the standard solution; if the deviation is in the set range, the serum in the purifying column is pumped by a first pump, and the purified serum is sent to a storage unit by switching of a first multi-channel direction selection valve; if the deviation is not within the set range, the serum in the purifying column is purified continuously. The invention has the advantages of good reliability and the like.
Description
Technical Field
The invention relates to serum purification, in particular to a preparation method of blank serum.
Background
When UPLC-MS/MS analyzes human serum, the matrix effect seriously affects the analysis of the instrument, so when preparing a standard curve, human/animal serum is used as a matrix to be simultaneously analyzed. Some endogenous analytes are always in serum, so that the standard curve is difficult to prepare, and analysis by using background subtraction causes overlarge deviation of low-concentration analysis and unreliable data.
Disclosure of Invention
In order to overcome the defects in the prior art scheme, the invention provides a blank serum preparation device.
The purpose of the invention is realized by the following technical scheme:
the preparation method of the blank serum comprises the following steps:
allowing the serum to enter a purifying column to remove endogenous compounds in the serum;
the first multi-channel direction selection valve is switched, the first pump pumps, and purified blood is discharged out of the purification column and enters the first quantitative ring; the first pump is sequentially connected with the first quantitative ring and the common end of the first multi-channel direction selection valve;
the first multi-channel direction selection valve is switched with the multi-way valve, the first pump pushes purified serum in the first quantitative ring, the purified serum enters the second quantitative ring, and two ends of the second quantitative ring are respectively connected with ports of the multi-way valve;
the multi-way valve is switched, the carrier liquid pushes the purified serum in the second quantitative ring to enter an analyzer, and the analyzer gives parameters of the purified serum;
calculating the deviation between the parameters and the parameters of the standard solution;
if the deviation is in a set range, the serum in the purifying column is pumped by the first pump, and the purified serum is sent to a storage unit by switching of a first multi-channel direction selection valve;
and if the deviation is no longer within the set range, continuously purifying the serum in the purifying column.
Compared with the prior art, the invention has the beneficial effects that:
1. automation;
the purification, analysis and comparison of the serum are automatically completed, the endogenous substance content of the purified serum is judged according to the measurement result, if the endogenous substance content is qualified, the endogenous substance content is collected and stored, and if the endogenous substance content is not qualified, the purification process is carried out again, the whole process is automatically operated without human intervention;
after the filler in the purification column is saturated, the automatic elution and purification are carried out, so that the reutilization is realized;
2. effectively solves the problem that endogenous compounds such as water-soluble vitamins, bile acid, neurotransmitter and the like in the current clinical kit select blank matrixes.
Drawings
The disclosure of the present invention will become more readily understood with reference to the accompanying drawings. As is readily understood by those skilled in the art: these drawings are only for illustrating the technical solutions of the present invention and are not intended to limit the scope of the present invention. In the figure:
FIG. 1 is a schematic flow diagram of a method for preparing blank serum according to an embodiment of the present invention.
Detailed Description
Fig. 1 and the following description depict alternative embodiments of the invention to teach those skilled in the art how to make and reproduce the invention. Some conventional aspects have been simplified or omitted for the purpose of explaining the technical solution of the present invention. Those skilled in the art will appreciate that variations or substitutions from these embodiments will be within the scope of the invention. Those skilled in the art will appreciate that the features described below can be combined in various ways to form multiple variations of the invention. Thus, the present invention is not limited to the following alternative embodiments, but is only limited by the claims and their equivalents.
Example 1:
as shown in fig. 1, the preparation method of the blank serum according to the embodiment of the present invention comprises:
driven by a pump, enabling the serum to enter a purifying column to remove endogenous compounds in the serum;
the first multi-channel direction selection valve is switched, the first pump pumps, and purified blood is discharged out of the purification column and enters the first quantitative ring; the first pump is sequentially connected with the first quantitative ring and the common end of the first multi-channel direction selection valve;
the first multi-channel direction selection valve is switched with the multi-way valve, the first pump pushes purified serum in the first quantitative ring, the purified serum enters the second quantitative ring, and two ends of the second quantitative ring are respectively connected with ports of the multi-way valve;
the multi-way valve is switched, the carrier liquid pushes the purified serum in the second quantitative ring to enter an analyzer, and the analyzer gives parameters of the purified serum;
calculating the deviation between the parameters and the parameters of the standard solution;
if the deviation is in a set range, the serum in the purifying column is pumped by the first pump, and the purified serum is sent to a storage unit by switching of a first multi-channel direction selection valve;
and if the deviation is no longer within the set range, continuously purifying the serum in the purifying column.
In order to automatically obtain the parameters of the standard solution, further, the parameters of the standard solution are obtained by the following steps:
switching a first multi-channel direction selection valve, pumping by a first pump, and allowing the standard liquid to enter a first quantitative ring;
the first multi-channel direction selection valve and the multi-way valve are switched, and the first pump pushes the standard liquid in the first quantitative ring to enter the second quantitative ring;
and the multi-way valve is switched, the carrier liquid pushes the standard liquid in the second quantitative ring to enter an analyzer, and the analyzer gives parameters of the standard liquid.
In order to deliver the serum, the serum is further delivered by a peristaltic pump, and the serum is filtered and then delivered to a purification column.
In order to automatically push the liquid in the second quantitative ring into the analyzer, further, the carrier liquid adopts a solvent, and the mode for pushing the liquid in the second quantitative ring is as follows:
the second multi-channel direction selection valve is switched, and the second pump extracts the selected solvent;
and the second multi-way valve option valve and the multi-way valve are switched, and the second pump pushes out the sucked solvent, so that the liquid in the second quantitative ring is pushed to enter the analyzer.
In order to recycle the resin particles in the purification column, the resin particles in the purification column are discharged, and then are subjected to alkali liquor soaking, ethanol purification and pure water purification in sequence, and then are sent back to the purification column.
Example 2:
example of an application of the method for preparing blank serum according to example 1 of the present invention.
In the present application example, as shown in fig. 1, the blank serum is prepared by the following method:
under the drive of a peristaltic pump, allowing the serum to enter a purification column, removing endogenous compounds in the serum, and filling macroporous resin particles in the purification column;
the first multi-channel direction selection valve is switched, the first injection pump pumps, and purified blood is discharged out of the purification column and enters the first quantitative ring; the first injection pump is sequentially connected with the first quantitative ring and the common end of the first multi-channel direction selection valve;
the first multi-channel direction selection valve and the six-way valve are switched, the first injection pump pushes purified serum in the first quantitative ring, the purified serum enters the second quantitative ring, and two ends of the second quantitative ring are respectively connected with the port of the six-way valve;
the six-way valve is switched, carrier liquid pushes the purified serum in the second quantitative ring to enter an analyzer (sequentially enter a chromatographic column and a mass spectrometer), and the analyzer gives parameters of the purified serum;
switching a first multi-channel direction selection valve, sucking by a first injection pump, and allowing the standard solution to enter a first quantitative ring;
the first multi-channel direction selection valve and the six-way valve are switched, and the first injection pump pushes the standard liquid in the first quantitative ring to enter the second quantitative ring;
the six-way valve is switched, carrier liquid pushes the standard liquid in the second quantitative ring to enter an analyzer (sequentially enter a chromatographic column and a mass spectrometer), and the analyzer gives parameters of the standard liquid;
calculating the deviation between the parameters and the parameters of the standard solution;
if the deviation is in a set range, the serum in the purifying column is pumped by the first pump, and the purified serum is sent to a storage unit by switching of a first multi-channel direction selection valve, wherein the storage unit comprises a refrigerator and a container;
if the deviation is no longer within the set range, continuously purifying the serum in the purifying column;
in the above process, the carrier liquid is solvent, and the mode of pushing the liquid in the second quantitative ring is as follows:
the second multi-channel direction selection valve is switched, and the second injection pump extracts the selected solvent;
the second multi-way valve option valve and the six-way valve are switched, and the second injection pump pushes out the absorbed solvent, so that the liquid (purified serum or standard liquid) in the second quantitative ring is pushed into the analyzer;
with the lengthening of the serum purification time, the macroporous resin particles in the purification column are gradually saturated, at the moment, the resin particles in the purification column are discharged, and are subjected to alkali liquor soaking, ethanol purification and pure water purification in sequence, and then are sent back to the purification column.
Claims (7)
1. The preparation method of the blank serum comprises the following steps:
allowing the serum to enter a purifying column to remove endogenous compounds in the serum;
the first multi-channel direction selection valve is switched, the first pump pumps, and purified blood is discharged out of the purification column and enters the first quantitative ring; the first pump is sequentially connected with the first quantitative ring and the common end of the first multi-channel direction selection valve;
the first multi-channel direction selection valve is switched with the multi-way valve, the first pump pushes purified serum in the first quantitative ring, the purified serum enters the second quantitative ring, and two ends of the second quantitative ring are respectively connected with ports of the multi-way valve;
the multi-way valve is switched, the carrier liquid pushes the purified serum in the second quantitative ring to enter an analyzer, and the analyzer gives parameters of the purified serum;
calculating the deviation between the parameters and the parameters of the standard solution;
if the deviation is in a set range, the serum in the purifying column is pumped by the first pump, and the purified serum is sent to a storage unit by switching of a first multi-channel direction selection valve;
and if the deviation is no longer within the set range, continuously purifying the serum in the purifying column.
2. The method for preparing blank serum according to claim 1, wherein the parameters of the standard solution are obtained by:
switching a first multi-channel direction selection valve, pumping by a first pump, and allowing the standard liquid to enter a first quantitative ring;
the first multi-channel direction selection valve and the multi-way valve are switched, and the first pump pushes the standard liquid in the first quantitative ring to enter the second quantitative ring;
and the multi-way valve is switched, the carrier liquid pushes the standard liquid in the second quantitative ring to enter an analyzer, and the analyzer gives parameters of the standard liquid.
3. The method for preparing blank serum according to claim 1, wherein the serum is delivered by a peristaltic pump, and the serum is filtered and then delivered to a purification column.
4. The method of claim 1, wherein the analyzer comprises a chromatographic column and a mass spectrometer.
5. The method for preparing blank serum according to claim 1, wherein macroporous resin particles are filled in the purification column.
6. The method for preparing blank serum according to claim 1, wherein the carrier liquid is a solvent, and the method for pushing the liquid in the second quantitative ring is as follows:
the second multi-channel direction selection valve is switched, and the second pump extracts the selected solvent;
and the second multi-way valve option valve and the multi-way valve are switched, and the second pump pushes out the sucked solvent, so that the liquid in the second quantitative ring is pushed to enter the analyzer.
7. The method for preparing blank serum according to claim 5, wherein the resin particles in the purification column are discharged, subjected to alkali soaking, ethanol purification and pure water purification in sequence, and then returned to the purification column.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111654145.XA CN114324689A (en) | 2021-12-30 | 2021-12-30 | Preparation method of blank serum |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111654145.XA CN114324689A (en) | 2021-12-30 | 2021-12-30 | Preparation method of blank serum |
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| Publication Number | Publication Date |
|---|---|
| CN114324689A true CN114324689A (en) | 2022-04-12 |
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| CN202111654145.XA Pending CN114324689A (en) | 2021-12-30 | 2021-12-30 | Preparation method of blank serum |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH07185584A (en) * | 1993-12-27 | 1995-07-25 | Nippon Sharyo Seizo Kaisha Ltd | Waste water treatment device |
| US5468643A (en) * | 1991-08-28 | 1995-11-21 | The United States Of America As Represented By The Department Of Health And Human Services | Switching valve system for direct biological sample injection for LC analysis |
| JP2007135908A (en) * | 2005-11-18 | 2007-06-07 | Jms Co Ltd | Blood purifying device |
| CN108717083A (en) * | 2018-01-18 | 2018-10-30 | 苏州和合医学检验有限公司 | The on-line solid phase extraction analysis method and system of diazepam and Clozapine detection in human serum |
| CN110873753A (en) * | 2018-08-30 | 2020-03-10 | 中国科学院大连化学物理研究所 | Enrichment method for separating gas-phase free propofol in whole blood sample |
-
2021
- 2021-12-30 CN CN202111654145.XA patent/CN114324689A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5468643A (en) * | 1991-08-28 | 1995-11-21 | The United States Of America As Represented By The Department Of Health And Human Services | Switching valve system for direct biological sample injection for LC analysis |
| JPH07185584A (en) * | 1993-12-27 | 1995-07-25 | Nippon Sharyo Seizo Kaisha Ltd | Waste water treatment device |
| JP2007135908A (en) * | 2005-11-18 | 2007-06-07 | Jms Co Ltd | Blood purifying device |
| CN108717083A (en) * | 2018-01-18 | 2018-10-30 | 苏州和合医学检验有限公司 | The on-line solid phase extraction analysis method and system of diazepam and Clozapine detection in human serum |
| CN110873753A (en) * | 2018-08-30 | 2020-03-10 | 中国科学院大连化学物理研究所 | Enrichment method for separating gas-phase free propofol in whole blood sample |
Non-Patent Citations (1)
| Title |
|---|
| 周从亚等: "液相色谱－ 离子淌度差分质谱法测定人血清中睾酮及雄烯二酮的浓度", 中国临床药理学杂志, vol. 36, no. 12, pages 1702 * |
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