CN116148391A - Blood sample preservation device, application thereof and blood sample detection method - Google Patents
Blood sample preservation device, application thereof and blood sample detection methodDownload PDFInfo
- Publication number
- CN116148391A CN116148391ACN202310217663.8ACN202310217663ACN116148391ACN 116148391 ACN116148391 ACN 116148391ACN 202310217663 ACN202310217663 ACN 202310217663ACN 116148391 ACN116148391 ACN 116148391A
- Authority
- CN
- China
- Prior art keywords
- blood sample
- adsorbent
- desiccant
- storage device
- container
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
- 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
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/10—Preservation of living parts
- A01N1/12—Chemical aspects of preservation
- A01N1/122—Preservation or perfusion media
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/10—Preservation of living parts
- A01N1/14—Mechanical aspects of preservation; Apparatus or containers therefor
- A01N1/142—Apparatus
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/4044—Concentrating samples by chemical techniques; Digestion; Chemical decomposition
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/405—Concentrating samples by adsorption or absorption
- 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
- 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
- G01N30/08—Preparation using an enricher
- 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
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
- 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
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
- 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
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- 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
- 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
- G01N2030/042—Standards
- G01N2030/047—Standards external
- 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
- 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
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
- G01N2030/8813—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample biological materials
- G01N2030/8822—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample biological materials involving blood
- 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
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
- G01N2030/884—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample organic compounds
Landscapes
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Dentistry (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Engineering & Computer Science (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Description
Translated fromChinese技术领域Technical Field
本发明涉及血液检测技术领域，尤其是涉及一种血液样品保存装置及其应用、血液样品的检测方法。The present invention relates to the technical field of blood testing, and in particular to a blood sample storage device and application thereof, and a blood sample testing method.
背景技术Background Art
在临床上、全血根据收集的条件，分成不抗凝和抗凝两种。同时，不抗凝分离出的上层黄色的液体称之为血清；抗凝分离出的上层黄色的液体称之为血浆。标本的存放温度及时间，血清、血浆及细胞分离的方法步骤也是分析前影响检验结果的重要因素。抽血后，血细胞因代谢需要，要消耗掉部分营养成分，故对这些成分进行测定时，需及时地离心以分离掉细胞成分。但是，对于大多数需要检测药物浓度的血清或者血浆样本需要在被分离后，放置-20℃以下条件下的低温冰箱进行储存，对于需要进行PK/PD的血清样本则需要在使用干冰在更低温条件下进行运输至检测中心进行检测，才能获得有效的临床数据。在这一过程中，不可避免需要花费大量的经济成本、时间成本、人力成本，从而保证项目的有效进行。In clinical practice, whole blood is divided into two types: non-anticoagulated and anticoagulated according to the conditions of collection. At the same time, the upper yellow liquid separated by non-anticoagulation is called serum; the upper yellow liquid separated by anticoagulation is called plasma. The storage temperature and time of the specimen, and the method and steps of serum, plasma and cell separation are also important factors affecting the test results before analysis. After blood is drawn, blood cells consume some nutrients due to metabolic needs. Therefore, when these components are measured, centrifugation is required in time to separate the cell components. However, for most serum or plasma samples that need to detect drug concentrations, they need to be stored in a low-temperature refrigerator below -20°C after separation. For serum samples that need to be tested for PK/PD, they need to be transported to the testing center for testing under lower temperature conditions using dry ice to obtain effective clinical data. In this process, it is inevitable to spend a lot of economic costs, time costs, and labor costs to ensure the effective implementation of the project.
现有血液样品的保存主要以低温储存为主，但存在如下缺陷：The current storage of blood samples is mainly based on low-temperature storage, but it has the following defects:
1、使用超低温冰箱储存样本，运输样本需要使用大量干冰，经济成本增加；1. Use ultra-low temperature refrigerators to store samples, and transport samples requires a large amount of dry ice, which increases economic costs;
2、低温储存的血清/血浆样本虽然可以提高了分析物的稳定性，解决了室温稳定性的问题，但是在进行检测前，需要提前解冻复溶至室温后进行后续化处理，时间较长；另外，对于代谢快的药物，在冻融这一过程中无法完全避免药物在临床血清/血浆中降解。2. Although serum/plasma samples stored at low temperatures can improve the stability of the analytes and solve the problem of room temperature stability, they need to be thawed and reconstituted at room temperature before testing, which takes a long time. In addition, for drugs with rapid metabolism, it is impossible to completely avoid drug degradation in clinical serum/plasma during the freeze-thaw process.
有鉴于此，特提出本发明。In view of this, the present invention is proposed.
发明内容Summary of the invention
本发明的第一目的在于提供一种血液样品保存装置，以解决上述问题中的至少一种。A first object of the present invention is to provide a blood sample storage device to solve at least one of the above problems.
本发明的第二目的在于提供上述血液样品保存装置的应用。A second object of the present invention is to provide an application of the above-mentioned blood sample storage device.
本发明的第三目的在于提供一种血液样品的检测方法。The third object of the present invention is to provide a method for detecting a blood sample.
第一方面，本发明提供了一种血液样品保存装置，包括干燥剂、吸附剂、蛋白变性剂和容器；In a first aspect, the present invention provides a blood sample storage device, comprising a desiccant, an adsorbent, a protein denaturant, and a container;
所述吸附剂和蛋白变性剂混合后与干燥剂分区域填装于所述容器中；The adsorbent and protein denaturant are mixed and then filled into the container in different areas with the desiccant;
所述吸附剂用于吸附血液样品中的待分析物。The adsorbent is used for adsorbing the analyte in the blood sample.
作为进一步技术方案，所述干燥剂为中性干燥剂；As a further technical solution, the desiccant is a neutral desiccant;
所述中性干燥剂包括无水氯化钙、无水硫酸镁、无水硫酸钠和无水硫酸钙中的至少一种。The neutral desiccant includes at least one of anhydrous calcium chloride, anhydrous magnesium sulfate, anhydrous sodium sulfate and anhydrous calcium sulfate.
作为进一步技术方案，所述吸附剂包括C18填料和PSA(乙二胺-N-丙基硅烷化硅胶吸附剂)填料中的至少一种；As a further technical solution, the adsorbent includes at least one of a C18 filler and a PSA (ethylenediamine-N-propyl silanized silica gel adsorbent) filler;
所述蛋白变性剂包括硫酸铜和氯化铜中的至少一种。The protein denaturant includes at least one of copper sulfate and copper chloride.
作为进一步技术方案，所述干燥剂、吸附剂、蛋白变性剂的质量比为50:20:5-50:5:1。As a further technical solution, the mass ratio of the desiccant, adsorbent and protein denaturant is 50:20:5-50:5:1.
作为进一步技术方案，所述容器包括离心管。As a further technical solution, the container includes a centrifuge tube.
作为进一步技术方案，所述吸附剂和蛋白变性剂混合后填装于容器的中心，并与容器壁以干燥剂间隔。As a further technical solution, the adsorbent and protein denaturant are mixed and filled in the center of the container, and separated from the container wall by a desiccant.
作为进一步技术方案，所述血液样品包括血清和血浆；As a further technical solution, the blood sample includes serum and plasma;
所述待分析物包括帕罗西汀、米氮平、舍曲林、艾司西酞普兰、西酞普兰、米安色林、度洛西汀、阿戈美拉汀、文拉法辛、O-去甲文拉法辛、氟伏沙明、阿米替林、去甲替林、多虑平、去甲多虑平、氯米帕明、去甲氯米帕明、氟西汀、去甲氟西汀、曲唑酮、安非他酮和羟基安非他酮。The analytes include paroxetine, mirtazapine, sertraline, escitalopram, citalopram, mianserin, duloxetine, agomelatine, venlafaxine, O-desmethylvenlafaxine, fluvoxamine, amitriptyline, nortriptyline, doxepin, nordoxepin, clomipramine, norclomipramine, fluoxetine, norfluoxetine, trazodone, bupropion and hydroxybupropion.
第二方面，本发明提供了上述血液样品保存装置在血液样品保存或检测中的应用。In a second aspect, the present invention provides the use of the above-mentioned blood sample storage device in the storage or detection of blood samples.
第三方面，本发明提供了一种血液样品的检测方法，包括以下步骤：In a third aspect, the present invention provides a method for detecting a blood sample, comprising the following steps:
将血液样品置于所述的血液样品保存装置中保存，然后对血液样品中的待分析物进行分离提取和检测。The blood sample is placed in the blood sample storage device for storage, and then the analyte in the blood sample is separated, extracted and detected.
作为进一步技术方案，所述分离提取的方法包括萃取；所述萃取的萃取液包括乙腈；As a further technical solution, the separation and extraction method includes extraction; the extraction liquid includes acetonitrile;
所述检测的方法包括液相色谱检测和质谱检测。The detection method includes liquid chromatography detection and mass spectrometry detection.
与现有技术相比，本发明具有如下有益效果：Compared with the prior art, the present invention has the following beneficial effects:
1、能够快速干化处理血清/血浆样本，同时固定化吸附样本中的待分析物，待分析物由液态转化为固态储存方式，降低待分析物化学反应速率；1. It can quickly dry serum/plasma samples and immobilize and adsorb the analytes in the samples, converting the analytes from liquid to solid storage, thereby reducing the chemical reaction rate of the analytes;
2、填料中添加酶变性剂材料，可使血清/血浆中的酶变性失活，从而降低待分析物的酶代谢途径；2. Adding enzyme denaturant materials to the filler can denature and inactivate the enzymes in serum/plasma, thereby reducing the enzyme metabolic pathway of the analyte;
3、可提高血清/血浆样本中待分析物的稳定性，最终降低储存温度苛刻的难度，经发明人研究发现，在2-8℃条件下，本发明提供的保存装置能够实现血清/血浆样本不低于180天，不超过365天的保存；3. It can improve the stability of the analytes in the serum/plasma samples, and ultimately reduce the difficulty of the harsh storage temperature. The inventors have found that under the condition of 2-8°C, the storage device provided by the present invention can achieve the storage of serum/plasma samples for no less than 180 days and no more than 365 days;
4、临床样本储存过程已参与部分前处理过程，降低后续化前处理的难度。4. The clinical sample storage process has already participated in some pre-processing processes, reducing the difficulty of subsequent pre-processing.
附图说明BRIEF DESCRIPTION OF THE DRAWINGS
为了更清楚地说明本发明具体实施方式或现有技术中的技术方案，下面将对具体实施方式或现有技术描述中所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图是本发明的一些实施方式，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他的附图。In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.
图1为本发明实施例提供的流程图。FIG. 1 is a flow chart provided by an embodiment of the present invention.
具体实施方式DETAILED DESCRIPTION
下面将结合实施方式和实施例对本发明的实施方案进行详细描述，但是本领域技术人员将会理解，下列实施方式和实施例仅用于说明本发明，而不应视为限制本发明的范围。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。未注明具体条件者，按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者，均为可以通过市售购买获得的常规产品。The embodiments of the present invention will be described in detail below in conjunction with the embodiments and examples, but it will be appreciated by those skilled in the art that the following embodiments and examples are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. Based on the embodiments in the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative work are within the scope of protection of the present invention. If specific conditions are not specified, proceed according to normal conditions or the conditions recommended by the manufacturer. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be purchased commercially.
第一方面，本发明提供了一种血液样品保存装置，包括干燥剂、吸附剂、蛋白变性剂和容器；In a first aspect, the present invention provides a blood sample storage device, comprising a desiccant, an adsorbent, a protein denaturant, and a container;
所述吸附剂和蛋白变性剂混合后与干燥剂分区域填装于所述容器中；The adsorbent and protein denaturant are mixed and then filled into the container in different areas with the desiccant;
所述吸附剂用于吸附血液样品中的待分析物。The adsorbent is used for adsorbing the analyte in the blood sample.
需要说明的是，本发明中“分区域填装”是指将吸附剂和蛋白变性剂混合物与干燥剂填装于容器的不同区域，区别于将吸附剂、蛋白变性剂和干燥剂混合后再填装的方式，例如可以将吸附剂和蛋白变性剂混合物填装于容器的下层，将干燥剂填装于容器的上层；也可以将吸附剂和蛋白变性剂混合物填装于容器的上层，将干燥剂填装于容器的下层；还可以将吸附剂和蛋白变性剂混合物填装于容器的左侧，将干燥剂填装于容器的右侧；优选将吸附剂和蛋白变性剂填装于容器的中心，并与容器壁以干燥剂间隔，该方式能够使分析物更好的脱离有水的条件、不容易变质，因此保存效果更好。It should be noted that "divided area filling" in the present invention refers to filling the adsorbent and protein denaturant mixture and the desiccant in different areas of the container, which is different from the method of mixing the adsorbent, protein denaturant and desiccant and then filling them. For example, the adsorbent and protein denaturant mixture can be filled in the lower layer of the container, and the desiccant can be filled in the upper layer of the container; the adsorbent and protein denaturant mixture can be filled in the upper layer of the container, and the desiccant can be filled in the lower layer of the container; the adsorbent and protein denaturant mixture can be filled in the left side of the container, and the desiccant can be filled in the right side of the container; preferably, the adsorbent and protein denaturant are filled in the center of the container and separated from the container wall by the desiccant. This method can better keep the analyte away from water conditions and not easily deteriorate, so the preservation effect is better.
本发明利用除水剂、吸附剂、蛋白变性剂作为装填材料填充至离心管中待用，能够用于血清的保存，通过除水剂快速吸收样本中的水分，吸附剂能够快速吸附血清中的分析物，蛋白变性剂能使血清中的酶等蛋白质失活，从而可使样本快速干化保存，降低储存条件；并且在这一过程中，利用填料的特性，完成部分前处理操作，待需检测时，可减少前处理操作，提高样品处理的效率。The present invention utilizes dehydrating agent, adsorbent and protein denaturing agent as filling materials to fill into centrifuge tube for standby use, which can be used for serum preservation. The dehydrating agent can quickly absorb water in the sample, the adsorbent can quickly adsorb analytes in the serum, and the protein denaturing agent can inactivate enzymes and other proteins in the serum, so that the sample can be quickly dried and preserved, and the storage conditions are reduced. In this process, the characteristics of the filler are utilized to complete part of the pre-treatment operation, and when detection is required, the pre-treatment operation can be reduced, thereby improving the efficiency of sample processing.
在一些优选的实施方式中，所述干燥剂为中性干燥剂；In some preferred embodiments, the desiccant is a neutral desiccant;
所述中性干燥剂包括但不限于无水氯化钙、无水硫酸镁、无水硫酸钠和无水硫酸钙，或者采用本领域技术人员所熟知的其他中性干燥剂。The neutral desiccant includes, but is not limited to, anhydrous calcium chloride, anhydrous magnesium sulfate, anhydrous sodium sulfate and anhydrous calcium sulfate, or other neutral desiccant known to those skilled in the art.
在一些优选的实施方式中，所述吸附剂包括但不限于C18填料和PSA填料中的至少一种，能够实现对血液样品中待分析物的吸附。In some preferred embodiments, the adsorbent includes but is not limited to at least one of a C18 filler and a PSA filler, and is capable of adsorbing the analyte in the blood sample.
所述蛋白变性剂包括但不限于硫酸铜和氯化铜，或者采用本领域技术人员所熟知的其他蛋白变性剂。The protein denaturing agent includes, but is not limited to, copper sulfate and copper chloride, or other protein denaturing agents known to those skilled in the art.
在一些优选的实施方式中，所述干燥剂、吸附剂、蛋白变性剂的质量比例如可以为，但不限于50:20:5、50:10:5、50:5:5、50:5:2、50:5:1、50:10:1或50:20:1。In some preferred embodiments, the mass ratio of the desiccant, adsorbent and protein denaturant can be, for example, but not limited to, 50:20:5, 50:10:5, 50:5:5, 50:5:2, 50:5:1, 50:10:1 or 50:20:1.
在一些优选的实施方式中，所述容器包括但不限于离心管，或者本领域技术人员所熟知的用于血液样品保存的容器。In some preferred embodiments, the container includes but is not limited to a centrifuge tube, or a container for storing blood samples known to those skilled in the art.
在一些优选的实施方式中，所述血液样品包括血清和血浆；In some preferred embodiments, the blood sample includes serum and plasma;
所述待分析物包括帕罗西汀、米氮平、舍曲林、艾司西酞普兰、西酞普兰、米安色林、度洛西汀、阿戈美拉汀、文拉法辛、O-去甲文拉法辛、氟伏沙明、阿米替林、去甲替林、多虑平、去甲多虑平、氯米帕明、去甲氯米帕明、氟西汀、去甲氟西汀、曲唑酮、安非他酮和羟基安非他酮。The analytes include paroxetine, mirtazapine, sertraline, escitalopram, citalopram, mianserin, duloxetine, agomelatine, venlafaxine, O-desmethylvenlafaxine, fluvoxamine, amitriptyline, nortriptyline, doxepin, nordoxepin, clomipramine, norclomipramine, fluoxetine, norfluoxetine, trazodone, bupropion and hydroxybupropion.
第二方面，本发明提供了上述血液样品保存装置在血液样品保存或检测中的应用。In a second aspect, the present invention provides the use of the above-mentioned blood sample storage device in the storage or detection of blood samples.
本发明提供的血液样品保存装置，通过除水剂快速吸收样本中的水分，吸附剂能够快速吸附血清中的分析物，蛋白变性剂能使血清中的酶等蛋白质失活，从而可使样本快速干化保存，降低储存条件；并且在这一过程中，利用填料的特性，完成部分前处理操作，待需检测时，可减少前处理操作，提高样品处理的效率。The blood sample storage device provided by the present invention can quickly absorb moisture in the sample through a dehydrating agent, the adsorbent can quickly adsorb analytes in the serum, and the protein denaturing agent can inactivate enzymes and other proteins in the serum, so that the sample can be quickly dried and stored, reducing storage conditions; and in this process, the characteristics of the filler are used to complete part of the pre-treatment operations, and when testing is required, the pre-treatment operations can be reduced, thereby improving the efficiency of sample processing.
第三方面，本发明提供了一种血液样品的检测方法，包括以下步骤：In a third aspect, the present invention provides a method for detecting a blood sample, comprising the following steps:
将血液样品置于所述的血液样品保存装置中保存，然后对血液样品中的待分析物进行分离提取和检测。The blood sample is placed in the blood sample storage device for storage, and then the analyte in the blood sample is separated, extracted and detected.
本发明提供的血液样品的检测方法简单方便，能够准确实现血液样品中待分析物的检测。The blood sample detection method provided by the present invention is simple and convenient, and can accurately detect the analyte in the blood sample.
在一些优选的实施方式中，所述分离提取的方法包括萃取；所述萃取的萃取液包括乙腈。In some preferred embodiments, the separation and extraction method comprises extraction; and the extraction liquid comprises acetonitrile.
在一些优选的实施方式中，在超声的条件下进行萃取。In some preferred embodiments, the extraction is performed under ultrasonic conditions.
在一些优选的实施方式中，所述检测的方法包括但不限于液相色谱检测和质谱检测，或者采用本领域技术人员所熟知的其他检测方法。In some preferred embodiments, the detection method includes but is not limited to liquid chromatography detection and mass spectrometry detection, or other detection methods well known to those skilled in the art.
下面通过具体的实施例和对比例进一步说明本发明，但是，应当理解为，这些实施例仅仅是用于更详细地说明之用，而不应理解为用于以任何形式限制本发明。The present invention is further described below by specific examples and comparative examples. However, it should be understood that these examples are only used for more detailed description and should not be construed as limiting the present invention in any form.
实施例1Example 1
实验流程如图1所示。The experimental process is shown in Figure 1.
1、制备血液样品保存装置1. Preparation of blood sample storage device
称取无水CaCl2、无水MgSO4混合固体粉末各25mg混匀，装填至离心管底部；称取C18填料10mg、PSA填料10mg和蛋白变性剂硫酸铜2mg混匀，装填至离心管中心位置，与离心管壁以干燥剂间隔。Weigh 25 mg of anhydrous CaCl2 and anhydrous MgSO4 mixed solid powder, mix evenly, and fill it to the bottom of the centrifuge tube; weigh 10 mg of C18 filler, 10 mg of PSA filler and 2 mg of protein denaturant copper sulfate, mix evenly, and fill it to the center of the centrifuge tube, separating it from the wall of the centrifuge tube with a desiccant.
2、制备血清校准品、质控品溶液2. Preparation of serum calibrator and quality control solutions
(A)5％BSA溶液:按照5g/100mL的比例称取牛血清白蛋白、磷酸缓冲溶液置于250mL PP瓶中，按照0.4％的比例添加防腐剂Proclin 300，颠倒混匀后，置于超声波清洗器中超声混匀10min。(A) 5% BSA solution: Weigh bovine serum albumin and phosphate buffer solution at a ratio of 5 g/100 mL and place in a 250 mL PP bottle. Add the preservative Proclin 300 at a ratio of 0.4%. Mix by inversion and place in an ultrasonic cleaner for 10 min.
(B)取混合治疗药物标准品中间液和5％BSA溶液按照5：95的比例混合，得到系列校准品、质控品溶液，振荡混匀10min，浓度参照下表，备用。(B) Mix the intermediate solution of the mixed therapeutic drug standard and 5% BSA solution in a ratio of 5:95 to obtain a series of calibration and quality control solutions. Oscillate and mix for 10 minutes. The concentrations are based on the table below and are set aside.
注：表中数据的单位为ng/mL。Note: The unit of data in the table is ng/mL.
3、固定化处理样本3. Fixation of samples
采用上述制备得到的血液样品保存装置固定化处理校准品、质控品，步骤如下：The blood sample storage device prepared above is used to immobilize the calibrant and the quality control product, and the steps are as follows:
1)向血液样品保存装置中添加校准品或质控品50μL、吸附10min；1) Add 50 μL of calibrator or quality control product to the blood sample storage device and adsorb for 10 minutes;
2)将处理好的校准品或质控品放置2-8℃储存。2) Store the processed calibrators or quality control materials at 2-8°C.
4、质控品赋值与上机检测4. Assignment of quality control products and on-machine testing
(A)取校准品、质控品、待测临床样本后续化前处理后，吸取上清液100μL，进样上机、检测。(A) After subsequent pretreatment of calibrators, quality control products, and clinical samples to be tested, 100 μL of the supernatant was aspirated and injected into the instrument for testing.
(B)LC－MS参数(B) LC-MS parameters
质谱仪为杭州谱聚医疗科技有限公司生产的PreMed5200液相色谱串联质谱系统。The mass spectrometer is the PreMed5200 liquid chromatography tandem mass spectrometry system produced by Hangzhou PuJu Medical Technology Co., Ltd.
①液相色谱条件：①Liquid chromatography conditions:
色谱柱：艾杰尔C18；Chromatographic column: Aeger C18;
50×3.0mm，5μm色谱柱。50×3.0mm, 5μm column.
流动相A：0.1％甲酸、2mM甲酸铵水溶液超声10min，备用。Mobile phase A: 0.1% formic acid, 2 mM ammonium formate aqueous solution, sonicate for 10 min and set aside.
流动相B：0.1％甲酸、2mM甲酸铵甲醇溶液超声10min，备用。Mobile phase B: 0.1% formic acid, 2 mM ammonium formate methanol solution, sonicate for 10 min and set aside.
柱温：35℃。Column temperature: 35°C.
进样量：5μl(100微升定量环，微升进样模式，吸样针取样高度2mm)。Injection volume: 5 μl (100 μl quantitative loop, microliter injection mode, sampling needle sampling height 2 mm).
梯度洗脱：Gradient elution:
②质谱条件②Mass spectrometry conditions
电喷雾离子源(ESI)，正离子，MRM扫描模式。Electrospray ionization (ESI), positive ion, MRM scanning mode.
离子源参数：Ion source parameters:
质谱MRM参数：Mass spectrometry MRM parameters:
5、结果5. Results
按照上述装置处理样本，分别在180天与365天测试稳定性，去甲文拉法辛、米氮平、羟基安非他酮、曲唑酮、安非他酮、文拉法辛、艾司西酞普兰、西酞普兰、米安色林、多虑平、去甲多虑平、度洛西汀、帕罗西汀、阿米替林、去甲替林、氟西汀、去甲氟西汀、氟伏沙明、氯米帕明、舍曲林、阿戈美拉汀、去甲氯米帕明准确度均在85％-115％之间，稳定性满足方法学要求。另外，其他种类临床药物以及更长的稳定性效期未作验证。The samples were processed according to the above device, and the stability was tested for 180 days and 365 days respectively. The accuracy of desvenlafaxine, mirtazapine, hydroxybupropion, trazodone, bupropion, venlafaxine, escitalopram, citalopram, mianserin, doxepin, nordoxepin, duloxetine, paroxetine, amitriptyline, nortriptyline, fluoxetine, norfluoxetine, fluvoxamine, clomipramine, sertraline, agomelatine, and norclomipramine was between 85% and 115%, and the stability met the methodological requirements. In addition, other types of clinical drugs and longer stability periods were not verified.
最后应说明的是：以上各实施例仅用以说明本发明的技术方案，而非对其限制；尽管参照前述各实施例对本发明进行了详细的说明，本领域的普通技术人员应当理解：其依然可以对前述各实施例所记载的技术方案进行修改，或者对其中部分或者全部技术特征进行等同替换；而这些修改或者替换，并不使相应技术方案的本质脱离本发明各实施例技术方案的范围。Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein by equivalents. However, these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention.
Claims (10)
Translated fromChinesePriority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310217663.8ACN116148391A (en) | 2023-03-02 | 2023-03-02 | Blood sample preservation device, application thereof and blood sample detection method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310217663.8ACN116148391A (en) | 2023-03-02 | 2023-03-02 | Blood sample preservation device, application thereof and blood sample detection method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116148391Atrue CN116148391A (en) | 2023-05-23 |
Family
ID=86361824
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310217663.8APendingCN116148391A (en) | 2023-03-02 | 2023-03-02 | Blood sample preservation device, application thereof and blood sample detection method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116148391A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115713193A (en)* | 2022-04-20 | 2023-02-24 | 杭州谱聚医疗科技有限公司 | Sequence planning method for automatic operation of samples |
| CN117705974A (en)* | 2023-12-13 | 2024-03-15 | 杭州汉科生物科技有限公司 | Kit for screening pesticide and drug poisoning and application method thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4871675A (en)* | 1984-11-23 | 1989-10-03 | Jiri Coupek | Storage container of samples for analysis |
| US20050227269A1 (en)* | 2004-04-09 | 2005-10-13 | Research Think Tank, Inc. | Devices and methods for collection, storage and transportation of biological specimens |
| CN101743317A (en)* | 2007-05-16 | 2010-06-16 | 凡利安股份有限公司 | Devices and methods for reducing matrix effects |
| WO2017210218A1 (en)* | 2016-05-31 | 2017-12-07 | Siscapa Assay Technologies, Inc. | Device and methods for sample collection |
| CN111239282A (en)* | 2020-02-08 | 2020-06-05 | 宁波市疾病预防控制中心 | Method and kit for determining phenobarbital in blood and application |
| CN113820424A (en)* | 2021-09-27 | 2021-12-21 | 厦门市仙岳医院(厦门市精神卫生中心) | HPLC-MS/MS method for simultaneously determining concentration of 14 antidepressants in human plasma |
- 2023
- 2023-03-02CNCN202310217663.8Apatent/CN116148391A/enactivePending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4871675A (en)* | 1984-11-23 | 1989-10-03 | Jiri Coupek | Storage container of samples for analysis |
| US20050227269A1 (en)* | 2004-04-09 | 2005-10-13 | Research Think Tank, Inc. | Devices and methods for collection, storage and transportation of biological specimens |
| CN101743317A (en)* | 2007-05-16 | 2010-06-16 | 凡利安股份有限公司 | Devices and methods for reducing matrix effects |
| WO2017210218A1 (en)* | 2016-05-31 | 2017-12-07 | Siscapa Assay Technologies, Inc. | Device and methods for sample collection |
| CN111239282A (en)* | 2020-02-08 | 2020-06-05 | 宁波市疾病预防控制中心 | Method and kit for determining phenobarbital in blood and application |
| CN113820424A (en)* | 2021-09-27 | 2021-12-21 | 厦门市仙岳医院(厦门市精神卫生中心) | HPLC-MS/MS method for simultaneously determining concentration of 14 antidepressants in human plasma |
Non-Patent Citations (1)
| Title |
|---|
| 艾 贾维茨 等: "医学微生物学", 31 October 1983, 人民卫生出版社, pages: 105 - 106* |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115713193A (en)* | 2022-04-20 | 2023-02-24 | 杭州谱聚医疗科技有限公司 | Sequence planning method for automatic operation of samples |
| CN117705974A (en)* | 2023-12-13 | 2024-03-15 | 杭州汉科生物科技有限公司 | Kit for screening pesticide and drug poisoning and application method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN116148391A (en) | Blood sample preservation device, application thereof and blood sample detection method | |
| Keevil | The analysis of dried blood spot samples using liquid chromatography tandem mass spectrometry | |
| US7901943B2 (en) | Materials and methods for controlling isotope effects during fractionation of analytes | |
| US20130236977A1 (en) | Compositions and methods for plasma peptide analysis | |
| WO2022120752A1 (en) | Method for quantitative analysis of free amino acids in biological sample by liquid chromatography-tandem mass spectrometry | |
| Moore et al. | Characterization of an immobilized enzyme reactor for on-line protein digestion | |
| CN104374848B (en) | A kind of method that phenyl boric acid material is enriched with glycopeptide | |
| AU2002341862A1 (en) | Controlling isotope effects during fractionation of analytes | |
| CN111398480A (en) | Kit for simultaneously detecting triazole antifungal drugs and glycopeptide antibiotics and detection method thereof | |
| JP2014507657A (en) | Method and system for determining the presence or amount of testosterone in a sample | |
| US20240350944A1 (en) | Separating and quantifying unbound target analytes from biological samples | |
| CN111983244A (en) | A kind of detection method and detection kit of four vitamins in dried blood spot | |
| US20070166767A1 (en) | Method and apparatus for mass spectrometric immunoassay analysis of specific biological fluid proteins | |
| CN113834888B (en) | Accurate detection method and kit for 4 ceramides in blood | |
| CN119125382A (en) | Method for detecting PTH1-84 by liquid chromatography-mass spectrometry | |
| Wang et al. | Mass spectrometry approaches for detection and determination of prostaglandins from biological samples | |
| De Cristofaro et al. | Decoding the Challenges: navigating Intact Peptide Mass Spectrometry-Based Analysis for Biological Applications | |
| CN115656400A (en) | Method for detecting 11-dehydrothromboxane B in urine 2 Liquid chromatography-tandem mass spectrometry method and kit | |
| CN114755348A (en) | Method for simultaneously detecting contents of 20 medicines and metabolites thereof | |
| CN118501313B (en) | Angiotensin detection kit and preparation method thereof | |
| CN116858948A (en) | Quantitative analysis method of plasma or serum protein based on liquid mass spectrometry technology | |
| CN111965369A (en) | Kit and method for detecting angiotensin I in blood plasma | |
| CN114609261A (en) | A method for the detection of 25-hydroxyvitamin D in dried blood spots by HPLC-MS | |
| CN118465117B (en) | A method and kit for specifically detecting multiple tricyclic antidepressants and its application | |
| CN118243826B (en) | Metabolite combination for assessing risk of biliary tract locking of neonate and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |