CN110987565A - Method for quantitatively detecting serum elements by inductively coupled plasma mass spectrometry - Google Patents

Method for quantitatively detecting serum elements by inductively coupled plasma mass spectrometry Download PDF

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CN110987565A
CN110987565A CN201911356784.0A CN201911356784A CN110987565A CN 110987565 A CN110987565 A CN 110987565A CN 201911356784 A CN201911356784 A CN 201911356784A CN 110987565 A CN110987565 A CN 110987565A
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冯振
景叶松
弭兆元
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Shandong Ying Sheng Biotechnology Co ltd
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Abstract

The invention discloses a method for quantitatively detecting serum elements by an inductively coupled plasma mass spectrometry, which comprises the steps of preprocessing a sample tube, and quantitatively measuring the serum elements by adopting the preprocessed sample tube, wherein the preprocessing step of the sample tube is as follows: and (3) soaking the sample tube in a nitric acid solution with the volume fraction of 1.5-5.5%, and cleaning and drying the soaked sample tube. The method can effectively reduce the sample background value in the experiment, and has the characteristics of simple operation, high accuracy, low detection limit and the like.

Description

Method for quantitatively detecting serum elements by inductively coupled plasma mass spectrometry
Technical Field
The invention belongs to the technical field of serum element analysis, and relates to a method for quantitatively detecting serum elements by inductively coupled plasma mass spectrometry (ICP-MS).
Background
The information in this background section is only for enhancement of understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.
The elements in human body can be divided into macroelements, microelements and toxic elements, wherein the macroelements are large in proportion in the human body, are required by the organism in large quantity, and are necessary elements for the organism, such as lithium, sodium, magnesium, phosphorus, potassium, calcium and the like; the trace elements are low in amount in a human body, but are also essential elements for maintaining life activities, such as chromium, manganese, iron, cobalt, nickel, copper, zinc, selenium and the like; toxic elements mainly refer to elements which are toxic to organisms (human bodies) and have no biological function, such as arsenic, molybdenum, cadmium, mercury, thallium, lead and the like. The environment where people are located and the food eaten by people influence the content of elements in human bodies, so that the detection of the content of the elements in the human bodies can help people to monitor the health problems and provide a basis for diagnosis and treatment of clinical element poisoning and the like.
At present, the hospital mainly adopts an Atomic Absorption Spectroscopy (AAS) method for detecting the content of elements in a blood sample. However, the AAS method has the advantages of low analysis speed, narrow linear range, easy interference of main components when measuring trace elements in a complex matrix sample, low precision and the like in a high-background low-content sample measurement task, and the inductively coupled plasma mass spectrometry (ICP-MS) has the advantages of high analysis speed, simultaneous measurement of multiple elements, wide linear range, high precision, good accuracy, low detection limit and the like, is suitable for trace element and trace element analysis, and is more suitable for the detection of multiple elements in human serum compared with the AAS method. The ICP-MS pretreatment method comprises a microwave digestion method, a high-pressure digestion method, a direct dilution method and the like, and compared with the microwave digestion method, the dilute nitric acid direct dilution method has the advantages of simple and quick operation steps, greatly improved pretreatment efficiency, shortened pretreatment time, avoidance of some potential pollution caused by microwave digestion to a certain extent, and capability of meeting the analysis requirements of clinical samples.
However, the research of the inventor of the present invention finds that the ICP-MS, even if the pretreatment is performed by a direct digestion method, is still interfered by some external conditions due to its high sensitivity, so that the background value of the trace elements measured in the sample is too high, thereby affecting the measurement result of the trace elements in the serum, increasing the experimental error, and affecting the accurate quantification of the trace elements.
Disclosure of Invention
In order to solve the defects of the prior art, the invention aims to provide the method for quantitatively detecting the serum elements by the inductively coupled plasma mass spectrometry, which can effectively reduce the sample background value in the experiment and has the characteristics of simple operation, high accuracy, low detection limit and the like.
In order to achieve the purpose, the technical scheme of the invention is as follows:
a method for quantitatively detecting serum elements by inductively coupled plasma mass spectrometry comprises the steps of preprocessing a sample tube, and quantitatively measuring the serum elements by adopting the preprocessed sample tube, wherein the preprocessing process of the sample tube comprises the following steps: and (3) soaking the sample tube in a nitric acid solution with the volume fraction of 1.5-5.5%, and cleaning and drying the soaked sample tube.
The inventor of the invention discovers that the pretreatment of the sample tube influences the background value of the sample in the experimental process, and the experiment discovers that the background value of the sample can be obviously reduced after the sample tube is soaked by a nitric acid solution with the volume fraction of 1.5-5.5%, so that the accuracy and the sensitivity of the quantitative detection of the serum element by the inductively coupled plasma mass spectrometry are improved.
The invention has the beneficial effects that:
the invention can effectively reduce the influence of impurities in the experimental consumables on the experimental result, so that the element measurement result, especially the measurement result of trace elements in serum, is more accurate, the operation method is simple, 21 elements in serum can be detected at one time, and a method is provided for clinical element detection.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this specification, are included to provide a further understanding of the invention, and are incorporated in and constitute a part of this specification, illustrate exemplary embodiments of the invention and together with the description serve to explain the invention and not to limit the invention.
FIG. 1 is a calibration curve for Al in an example of the present invention;
FIG. 2 is a calibration curve of Ca in the example of the present invention;
FIG. 3 is a calibration curve for Cd in an embodiment of the present invention;
FIG. 4 is a calibration curve of Co in an example of the present invention;
FIG. 5 is a calibration curve of Cr in an embodiment of the present invention;
FIG. 6 is a calibration curve for Cu in an example of the present invention;
FIG. 7 is a calibration curve for Fe in an example of the present invention;
FIG. 8 is a calibration curve for Hg in an example of the present invention;
FIG. 9 is a calibration curve for K in an example of the present invention;
FIG. 10 is a calibration curve for Li in an example of the present invention;
FIG. 11 is a calibration curve for Mg in an example of the present invention;
FIG. 12 is a calibration curve of Mn in the example of the present invention;
FIG. 13 is a calibration curve of Mo in the example of the present invention;
FIG. 14 is a calibration curve for Na in an example of the present invention;
FIG. 15 is a calibration curve for Ni in an example of the present invention;
FIG. 16 is a calibration curve for P in an example of the present invention;
FIG. 17 is a calibration curve of As in an example of the present invention;
FIG. 18 is a calibration curve of Se in an example of the present invention;
FIG. 19 is a calibration curve for Pb in the example of the present invention;
FIG. 20 is a calibration curve of Zn in the example of the present invention;
FIG. 21 is a calibration curve of Ti in examples of the present invention.
Detailed Description
It is to be understood that the following detailed description is exemplary and is intended to provide further explanation of the invention as claimed. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the invention. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.
In view of the problem of overhigh background value of trace elements measured by ICP-MS in the prior art, the invention provides a method for quantitatively detecting serum elements by inductively coupled plasma mass spectrometry.
The invention provides a method for quantitatively detecting serum elements by an inductively coupled plasma mass spectrometry, which comprises the steps of preprocessing a sample tube, and quantitatively measuring the serum elements by adopting the preprocessed sample tube, wherein the preprocessing step of the sample tube is as follows: and (3) soaking the sample tube in a nitric acid solution with the volume fraction of 1.5-5.5%, and cleaning and drying the soaked sample tube.
Experiments show that after the sample tube is soaked by a nitric acid solution with the volume fraction of 1.5-5.5%, the background value of the sample can be obviously reduced, so that the accuracy and the sensitivity of the inductively coupled plasma mass spectrometry on the quantitative detection of the serum elements are improved.
In one or more embodiments of this embodiment, the soaking time is not less than 30 min. Experiments show that the signal intensity of the blank sample of the sample tube is further reduced along with the increase of the soaking time, and the reduced signal intensity is not greatly different when the soaking time is more than 30min (including 30 min). In order to reduce the detection time and the detection cost, the sample tube can be soaked for 30 min.
The embodiment of using distilled water such as ouahaha drinking water, homemade purified water, and distilled water of Chen and Chen who is washed after soaking finds that the 3 kinds of water have little difference in detecting element content52Cr is most affected by blank background value, resulting in52The Cr measurement result is always inaccurate52Cr detection is more accurate, and in one or more examples of this embodiment, the soaked sample tube is cleaned with Chen's distilled water.
In one or more embodiments of this embodiment, the drying time is 1-4 hours.
In one or more examples of this embodiment, the element in the test serum is one or more of lithium, sodium, magnesium, aluminum, phosphorus, potassium, calcium, chromium, manganese, iron, cobalt, nickel, copper, zinc, arsenic, selenium, molybdenum, cadmium, mercury, thallium, and lead.
In one or more embodiments of this embodiment, the serum sample is placed in a pretreated sample tube, and a diluent is added to prepare the sample, wherein the volume fraction of the diluent is not higher than 10% of the nitric acid solution. When the volume fraction of the nitric acid solution is 1%, the detection effect is better.
In the series of examples, the volume ratio of the serum sample to the nitric acid solution is 0.1: 3.9-4.1.
In one or more embodiments of the present disclosure, a serum sample solution in a sample tube after pretreatment is brought into a plasma flame by a carrier gas in the form of aerosol, and is evaporated, decomposed, excited, and ionized to form charged ions, interference of polyatomic ions is removed by a collision reaction cell, then ions with different mass-to-nuclear ratios (m/z) are selected to pass through the collision reaction cell to detect the intensity of an ion, and the intensity of an element is analyzed and calculated, where the abundance intensity of the ion is proportional to the content of the element. The instrument quantitatively determines the calibration curve, draws a standard curve of each element, and calculates the content of lithium, sodium, magnesium, aluminum, phosphorus, potassium, calcium, chromium, manganese, iron, cobalt, nickel, copper, zinc, arsenic, selenium, molybdenum, cadmium, mercury, thallium and lead in the serum sample.
In one or more embodiments of this embodiment, the calibration solution is formulated as: and diluting the element standard mother liquor into a standard solution by adopting a diluent. In this series of examples, the dilution used in the calibration solution preparation process was the same as the dilution used in the sample preparation process.
In this series of examples, the diluent in the preparation of the internal standard solution was the same as the diluent in the preparation of the sample.
In order to make the technical solutions of the present invention more clearly understood by those skilled in the art, the technical solutions of the present invention will be described in detail below with reference to specific embodiments.
Examples
First, using the instrument and the instrument condition
The apparatus used was: a seemer fly iCAPQ plasma mass spectrometer.
The KED mode is selected as the instrument mode, and the collision gas is He.
Before the instrument is used, tuning and calibrating the instrument by using tuning liquid and correction liquid, wherein the instrument parameters are as follows:
forward power (Forward power): 1550V.
Nebulizer flow rate (Nebuliser flow): 1.02 L.min-1。
Helper flow rate (auxiary flow): 0.8 L.min-1。
Cooling gas flow rate (Cool flow): 14 L.min-1。
Sampling depth (Sampling depth): 158 nm.
Collision voltage (KED voltage): 3V.
The measured elements and the mass numbers thereof are as follows:
element(s) Mass number Element(s) Mass number
Li
7 Co 59
Na 23 Ni 60
Mg 24 Cu 63
Al 27 Zn 66
P 31 As 75
K 39 Se 78
Ca 44 Mo 95
Cr 52 Cd 111
Mn 55 Hg 202
Fe 57 Ti 205
Pb 208
Secondly, consumable cleaning process: respectively using different volume ratios: 0%, 0.1%, 1%, 2%, 5% dilute nitric acid, different soaking times: 0min, 15min, 30min, 1h, 2h, 4h, different washing water: the water purification method comprises the following steps of (1) Wahaha drinking water, Chechen distilled water and purified water prepared by a water purifier of the company (the water meets the requirements of YY/T1244-2014 purified water for in-vitro diagnostic reagents), and drying for different times: ICP-MS sample tubes (Thermo Scientific, USA) were cleaned and dried in four different ways, 1h, 2h and 4 h.
Thirdly, preparing a blank sample: 4mL of 1% dilute nitric acid solution was placed in the washed and dried sample tube as a blank sample.
Fourthly, preparing a serum sample: adding 100 μ L of serum sample into the dried sample tube, and adding 4mL of diluent, i.e. 1% HNO3And (3) solution.
Fifthly, preparing a standard solution: 1% HNO was used3The solution is used for diluting a standard stock solution containing 21 elements, wherein the standard stock solution is a mixed standard working solution with the content range of the elements in human serum as reference, major elements such as sodium, magnesium, phosphorus, potassium and calcium being diluted to ppm (mg/L), trace elements such as chromium, manganese, iron, cobalt, nickel, copper, zinc and selenium and toxic elements such as arsenic, molybdenum, cadmium, mercury, thallium and lead being diluted to ppb (mu g/L), and the preparation and dilution of the standard solution are carried out in a dried sample tube.
Sixthly, preparing an internal standard solution: choose to use45Sc、89Y、103Rh、159Tb、165Ho、115In、209Taking Bi as an internal standard, taking an internal standard mother solution, putting the internal standard mother solution into a dried sample tube, and using 1% of HNO3A mixed standard solution diluted to 10 ppb.
The background value of the blank sample was determined under the instrument conditions of the one above.
After the sample tubes are cleaned by the dilute nitric acid with different volume ratios, 4mL of blank samples are added and directly operated on a computer to determine the signal intensity, namely the blank background value, each sample is determined for 3 times, the obtained result is an average value of three times, and the result is shown in the following table 1:
TABLE 1 influence of signal intensity of blank samples after cleaning sample tubes with dilute nitric acid of different volume ratios
Figure BDA0002336143760000061
As can be seen from Table 1, the signal intensity of the blank sample in the experiment is reduced by using nitric acid to clean the sample tube, and the cleaning effect of 2% and 5% nitric acid is better through comparison, so that from the economic point of view, the nitric acid with the volume ratio of 2% can be selected to clean the sample tube.
After the sample tube is soaked in 2% dilute nitric acid for different time, 4mL of blank sample is added and directly operated on a computer to determine the blank background value, each sample is determined for 3 times, the obtained result is an average value of three times, and the result is as shown in the following table 2:
TABLE 2 influence of signal intensity of blank samples after soaking in 2% dilute nitric acid for various periods of time
Figure BDA0002336143760000071
It can be seen from table 2 that after the sample tube is soaked in 2% dilute nitric acid for a period of time, the signal intensity of the blank sample of the sample tube is further reduced, and the reduced signal intensities in the soaking times of 30min, 1h, 2h and 4h are not much different, so that 30min can be selected to soak the sample tube in order to save time.
The elements contained in the water also influence the signal intensity of a blank sample and influence the background value of the sample, thereby influencing the accuracy of the determination, so that the invention determines 3 different kinds of water: the water meets the requirements of YY/T1244-2014 'purified water for in-vitro diagnostic reagents' on element signal intensity in the Wahaha drinking water, the Chechen distilled water and the purified water prepared by the water purifier of the company. After soaking the sample tube in 2% dilute nitric acid for 30min, respectively washing the sample tube with 3 kinds of water, respectively preparing a blank sample in the corresponding water-washed sample tube with 3 kinds of water, respectively, measuring each sample for 3 times, and obtaining results as an average value of three times, wherein the results are as follows in table 3:
TABLE 3 influence of different water samples on the signal intensity of blank samples
Figure BDA0002336143760000081
As can be seen from the above table, the element contents in the 3 water samples are not greatly different, and the experiment of the invention52Cr is most affected by blank background value, resulting in52The Cr measurement result is always inaccurate, so the method is selected52Preparing diluent and cleaning a sample tube by using the Chen's distilled water with a lower Cr background value.
After the sample tube is cleaned, water needs to be drained to reduce errors, and the natural draining time is long, so that the method for drying the sample tube by using the oven saves time, and researches the influence of different drying times on the background value of the blank sample. As the sample tube is not completely dried within 30min, the drying time of the invention is 1h, 2h and 4 h. Soaking the sample tube in 2% dilute nitric acid for 30min, cleaning with Dreches distilled water, and drying the sample tube for different drying times. Preparing blank samples by using the Dreches distilled water in sample tubes after cleaning and drying, measuring each sample for 3 times, and obtaining results which are average values of three times, wherein the results are shown in the following table 4:
TABLE 4 influence of different drying times on the signal intensity of blank samples
Figure BDA0002336143760000091
After a series of cleaning, the background value of the blank sample in the sample tube is very low, and the drying time has little influence on the blank sample, so that the sample tube only needs to be dried without residual moisture.
Method detection limit
Soaking a sample tube for 30min by using 2% dilute nitric acid, cleaning the sample tube by using Chechen distilled water, then putting the sample tube into an oven for drying for 1h, adding a diluent prepared by Chechen distilled water, namely a blank sample, and carrying out 10 times of repeated measurement on the blank sample, wherein a calibration curve of 21 detected elements is shown in figures 1-21, the concentration value of the element corresponding to 3 times of standard deviation is a detection limit, and the detection limit of each element is shown in the following table 5:
TABLE 5 detection limits of the elements under the method
Element(s) Detection limit Element(s) Detection limit
7Li[ppb] 9.712 59Co[ppb] 0.072
23Na[ppm] 0.256 60Ni[ppb] 0.033
24Mg[ppm] 0.009 63Cu[ppb] 0.786
27Al[ppb] 1.344 66Zn[ppb] 0.842
31P[ppm] 0.026 75As[ppb] 0.043
39K[ppm] 0.604 78Se[ppb] 0.217
44Ca[ppm] 0.479 95Mo[ppb] 0.043
52Cr[ppb] 0.022 111Cd[ppb] 0.002
55Mn[ppb] 0.065 202Hg[ppb] 0.068
57Fe[ppb] 4.113 205Ti[ppb] 0.009
208Pb[ppb] 0.005
The method can reduce the background value of the sample, has low detection limit and good accuracy, and provides a method for measuring the element content by using ICP-MS in clinical experiments.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (10)

1. A method for quantitatively detecting serum elements by an inductively coupled plasma mass spectrometry is characterized by comprising the steps of preprocessing a sample tube and quantitatively measuring the serum elements by adopting the preprocessed sample tube, wherein the preprocessing step of the sample tube is as follows: and (3) soaking the sample tube in a nitric acid solution with the volume fraction of 1.5-5.5%, and cleaning and drying the soaked sample tube.
2. The method for quantitatively detecting serum elements by inductively coupled plasma mass spectrometry as claimed in claim 1, wherein the soaking time is not less than 30 min.
3. The method for quantitatively detecting serum elements by inductively coupled plasma mass spectrometry as claimed in claim 1, wherein the soaked sample tube is washed with Chen's distilled water.
4. The method for quantitatively detecting serum elements by inductively coupled plasma mass spectrometry as claimed in claim 1, wherein the drying time is 1-4 h.
5. The method of claim 1, wherein the serum is detected as one or more of lithium, sodium, magnesium, aluminum, phosphorus, potassium, calcium, chromium, manganese, iron, cobalt, nickel, copper, zinc, arsenic, selenium, molybdenum, cadmium, mercury, thallium, and lead.
6. The method for quantitatively detecting serum elements by inductively coupled plasma mass spectrometry as claimed in claim 1, wherein the serum sample is placed in a pretreated sample tube, and a diluent is added to prepare the sample, wherein the volume fraction of the diluent is not higher than 10% of that of a nitric acid solution.
7. The method for quantitatively detecting serum elements by inductively coupled plasma mass spectrometry as claimed in claim 6, wherein the volume ratio of the serum sample to the nitric acid solution is 0.1: 3.9-4.1.
8. The method as claimed in claim 1, wherein the method comprises introducing the serum sample solution in the sample tube into the plasma flame in the form of aerosol by carrier gas, evaporating, decomposing, exciting and ionizing to form charged ions, removing the interference of polyatomic ions by collision reaction, detecting the intensity of ions by selecting ions with different mass-to-nuclear ratios, analyzing and calculating the intensity of a certain element, quantitatively determining the calibration curve, drawing the standard curve of each element, and calculating the content of Li, Na, Mg, Al, P, K, Ca, Cr, Mn, Fe, Co, Ni, Cu, Zn, As, Se, Mo, Cd, Hg, Tl, Pb.
9. The method of claim 1, wherein the calibration solution is prepared by: diluting the element standard mother liquor into a standard solution by adopting a diluent;
preferably, the diluent in the calibration solution preparation process is the same as the diluent in the sample preparation process.
10. The method of claim 1, wherein the diluent in the preparation of the internal standard solution is the same as the diluent in the preparation of the sample.
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Application publication date: 20200410