CN108226271A - The detection method and kit of 19 kinds of elements of human body are detected simultaneously - Google Patents
The detection method and kit of 19 kinds of elements of human body are detected simultaneously Download PDFInfo
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Abstract
The present invention relates to a kind of while detect the detection method and kit of 19 kinds of elements of human body, detection method includes the following steps：19 kinds of elements are respectively：Zinc, strontium, selenium, lithium, aluminium, molybdenum, arsenic, iron, barium, copper, lead, mercury, nickel, calcium halophosphate activated by antimony andmanganese, cadmium, thallium, chromium and cobalt；(1) dilution：It is marked with containing 1.5 2.5mg/L gold and 0.1 2wt% aqueous solution of nitric acid of 0.05 0.2wt% Triton X-100s；Standard solution：It is formulated by adding in 19 kinds of element standards in the dilution；Inner mark solution：It is formulated by adding in internal standard element standard items in the dilution；(2) random time point urine is collected；The urine is through the diluted to get the sample to be tested；(3) it is detected using Plasma-Mass Spectroscopy ICP MS.Above-mentioned detection method can detect 19 kinds of elements in human body simultaneously, and the method for sampling is simple, sample is easily stored, pre-treatment is simple, high sensitivity, high specificity, detects flux height.
Description
Technical field
The present invention relates to technical field of analysis and detection, and plasma mass spectrometry (ICP-MS) is used simultaneously more particularly to a kind of
Detect the method and kit of 19 kinds of elements of human body.
Background technology
The organism that human body is made of various chemical elements.According to each element physiological action in human body, battalion can be divided into
Support element and toxic element.Nutrient, which refers to, special physiological function, and the element to maintaining human normal vital movement beneficial,
Such as：Calcium, phosphorus, magnesium, potassium, sodium, iron, zinc, copper, selenium etc..Toxic element refers to the micro member for existing and adverse effect will being caused to human body
Element common are heavy metal, such as arsenic, cadmium, lead, mercury.
The imbalance (nutrient deficiency or toxic element are excessively high) of element all can generate adverse effect to human body.2004
《Chinese residents nutrition and Health Situation》Investigation show although Chinese eat to become better and better, but there are still micro battalion
The shortage of element is supported, this shortage is often not noticeable, therefore is defined as " recessive hunger " by the World Health Organization (WHO).Country
What health and Family Planning Committee issued《Chinese residents nourishment and chronic disease status report (2015)》, show China resident
Diet structure is mostly there are unreasonable, and the minerals and vitamins such as calcium, iron, zinc, selenium do not reach recommended intake, and there are such battalion
Support the shortage of element.A variety of chronic diseases and inferior health are excessive related to toxicity in vivo element according to the study.In recent years due to undesirable
Life style and environmental pollution, we are at every moment likely to be exposed in the environment full of toxic element.So we need
It is to be understood that the situation of endotrophic prime element and toxic element, and assess whether that there are the shortage of nutrient, toxic elements to tire out
Situations such as product or poisoning.
The nutrient of detection low concentration and the common method of toxic element have at present：Atomic absorption method, Anodic Stripping
Voltammetry, atomic fluorescence method, neutron activation method, Ultraviolet Photometric Method, plasma emission spectrum (ICP-AES) and plasma mass spectrometry
(ICP-MS).Most common of which is ICP-AES and ICP-MS, but ICP-AES is directed to the detection limit of Partial Elements, sensitivity etc.
ICP-MS is below, and ICP-MS high sensitivities, specificity are stronger, can be described as the goldstandard of low concentration Element detection at present.
The concentration difference of nutrient and toxic element is larger in human body, it is impossible to while multiple element is detected, so when detecting usually
It needs to need to be placed on difference in an item detection, such as element of ppm grades of concentration and ppb ranks by the close element combinations of concentration
Project in.The sample type of detection human elementary is also more at present common are blood, twenty-four-hour urine liquid, hair etc..Blood
Sample pretreatment trouble and sampling complexity, the sample of hair are easier to contaminated and complex pretreatment, twenty-four-hour urine liquid sample and adopt
Sample is cumbersome to be needed the urine of collection 24 hours and transports cumbersome.Therefore we need to establish that a kind of method of sampling is simple, sample
The detection method that easily stored, pre-treatment is simple, high sensitivity, high specificity, detection flux are high.
Invention content
Based on this, the object of the present invention is to provide a kind of plasma mass spectrometry (ICP-MS) while 19 kinds of members of human body are detected
The method of element.
Specific technical solution is as follows：
Detection method that is a kind of while detecting 19 kinds of elements of human body, includes the following steps：
19 kinds of elements are respectively：Zinc, strontium, selenium, lithium, aluminium, molybdenum, arsenic, iron, barium, copper, lead, mercury, nickel, calcium halophosphate activated by antimony andmanganese, cadmium,
Thallium, chromium and cobalt；
(1) solution is prepared
Dilution：The 0.1-2wt% aqueous solution of nitric acid of the gold containing 1.5-2.5mg/L；
Standard solution：It is formulated by adding in 19 kinds of element standards in the dilution, wherein described 19
The concentration of kind of element is respectively：Zinc：5-2000 μ g/mL, strontium：2.5-1000μg/mL；Selenium 1.5-600 μ g/mL；Lithium：1.5-600μ
G/mL, aluminium：1.5-600 μ g/mL, molybdenum：0.75-300 μ g/mL, arsenic：0.75-300 μ g/mL, iron：0.75-300 μ g/mL, barium：
0.5-200 μ g/mL, copper：0.25-100 μ g/mL, lead：0.1-40 μ g/mL, mercury：0.1-40 μ g/mL, nickel：0.1-40 μ g/mL,
Antimony：0.05-20 μ g/mL, manganese：0.05-20 μ g/mL, cadmium：0.05-20 μ g/mL, thallium：0.025-10 μ g/mL, chromium：0.025-10μ
G/mL, cobalt：0.025-10μg/mL；
Inner mark solution：Be formulated by adding in internal standard element standard items in the dilution, the internal standard element for gallium and
Rhodium；
(2) sample to be tested
Acquire sample：Collect random time point urine；
Sample pre-treatments：The urine is through the diluted to get the sample to be tested；
(3) it detects
Inner mark solution is added in the standard solution, is detected using Plasma-Mass Spectroscopy ICP-MS, builds standard curve；
Wherein Mass Spectrometry Conditions are：
Add in inner mark solution in the sample to be tested, using Plasma-Mass Spectroscopy ICP-MS detect to get.
In wherein some embodiments, described the step of collecting sample, further includes：The urine of random time point is acquired, takes filter
Scraps of paper urine, dries, and obtains dry nappy sample.
In wherein some embodiments, the step of sample pre-treatments, further includes：The dry paper described in distillation water wash is urinated
Piece sample centrifuges to obtain the sample to be tested.
In wherein some embodiments, quality-control product solution is further included in the solution preparation steps：
It is formulated by adding in 19 kinds of element standards in the dilution, the quality-control product solution includes low dense
Spend three kinds of quality-control product solution, middle concentration quality-control product solution and high concentration quality-control product solution concentration；
The concentration of 19 kinds of elements described in the low concentration quality-control product solution is respectively：Zinc：120 μ g/mL, strontium：60μg/
mL；Selenium：40μg/mL；Lithium：20 μ g/mL, aluminium：20 μ g/mL, molybdenum：16 μ g/mL, arsenic：16 μ g/mL, 8 μ g/mL of iron, barium： 8μg/
ML, copper：4 μ g/mL, lead：1.6 μ g/mL, mercury：1.6 μ g/mL, nickel：1.6 μ g/mL, antimony：0.8 μ g/mL, manganese：0.8 μ g/mL, cadmium：
0.8 μ g/mL, thallium：0.4 μ g/mL, chromium：0.4 μ g/mL, cobalt：0.4μg/mL；
The concentration of 19 kinds of elements described in the middle concentration quality-control product solution is respectively：Zinc：240 μ g/mL, strontium：120μ
g/mL；Selenium：80μg/mL；Lithium：40 μ g/mL, aluminium：40 μ g/mL, molybdenum：32 μ g/mL, arsenic：32 μ g/mL, iron：16 μ g/mL, barium：16μ
G/mL, copper：8 μ g/mL, lead：3.2 μ g/mL, mercury：3.2 μ g/mL, nickel：3.2 μ g/mL, antimony：1.6 μ g/mL, manganese：1.6 μ g/mL,
Cadmium：1.6 μ g/mL, thallium：0.8 μ g/mL, chromium：0.8 μ g/mL, cobalt：0.8μg/mL；
The concentration of 19 kinds of elements described in the high concentration quality-control product solution is respectively：Zinc：600 μ g/mL, strontium：300μ
g/mL；Selenium：200μg/mL；Lithium：100 μ g/mL, aluminium：100 μ g/mL, molybdenum：80 μ g/mL, arsenic：80 μ g/mL, iron：40 μ g/mL, barium：
40 μ g/mL, copper：20 μ g/mL, lead：8 μ g/mL, mercury：8 μ g/mL, nickel：8 μ g/mL, antimony：4 μ g/mL, manganese：4 μ g/mL, cadmium：4μg/
ML, thallium：2 μ g/mL, chromium：2 μ g/mL, cobalt：2μg/mL.
The present invention also provides a kind of while detect the kits of 19 kinds of elements of human body, including dilution, standard solution with
And inner mark solution；
The dilution is the 0.1-2wt% aqueous solution of nitric acid of the gold containing 1.5-2.5mg/L；
The standard solution：It is formulated by adding in 19 kinds of element standards in the dilution, wherein described
The concentration of 19 kinds of elements is respectively：Zinc：5-2000 μ g/mL, strontium：2.5-1000μg/mL；Selenium 1.5-600 μ g/mL；Lithium：1.5-
600 μ g/mL, aluminium：1.5-600 μ g/mL, molybdenum：0.75-300 μ g/mL, arsenic：0.75-300 μ g/mL, iron：0.75-300 μ g/mL,
Barium：0.5-200 μ g/mL, copper：0.25-100 μ g/mL, lead：0.1-40 μ g/mL, mercury：0.1-40 μ g/mL, nickel：0.1-40μg/
ML, antimony：0.05-20 μ g/mL, manganese：0.05-20 μ g/mL, cadmium：0.05-20 μ g/mL, thallium：0.025-10 μ g/mL, chromium：0.025-
10 μ g/mL, cobalt：0.025-10μg/mL；
The inner mark solution：It is formulated by adding in internal standard element standard items in the dilution, the internal standard element is
Gallium and rhodium, a concentration of 1000 μ g/mL of gallium, a concentration of 1000 μ g/mL of rhodium.
In wherein some embodiments, the kit further includes quality-control product solution：
It is formulated by adding in 19 kinds of element standards in the dilution, the quality-control product solution includes low dense
Spend three kinds of quality-control product solution, middle concentration quality-control product solution and high concentration quality-control product solution concentration；
The concentration of 19 kinds of elements described in the low concentration quality-control product solution is respectively：Zinc：120 μ g/mL, strontium：60μg/
mL；Selenium：40μg/mL；Lithium：20 μ g/mL, aluminium：20 μ g/mL, molybdenum：16 μ g/mL, arsenic：16 μ g/mL, 8 μ g/mL of iron, barium： 8μg/
ML, copper：4 μ g/mL, lead：1.6 μ g/mL, mercury：1.6 μ g/mL, nickel：1.6 μ g/mL, antimony：0.8 μ g/mL, manganese：0.8 μ g/mL, cadmium：
0.8 μ g/mL, thallium：0.4 μ g/mL, chromium：0.4 μ g/mL, cobalt：0.4μg/mL；
The concentration of 19 kinds of elements described in the middle concentration quality-control product solution is respectively：Zinc：240 μ g/mL, strontium：120μ
g/mL；Selenium：80μg/mL；Lithium：40 μ g/mL, aluminium：40 μ g/mL, molybdenum：32 μ g/mL, arsenic：32 μ g/mL, iron：16 μ g/mL, barium：16μ
G/mL, copper：8 μ g/mL, lead：3.2 μ g/mL, mercury：3.2 μ g/mL, nickel：3.2 μ g/mL, antimony：1.6 μ g/mL, manganese：1.6 μ g/mL,
Cadmium：1.6 μ g/mL, thallium：0.8 μ g/mL, chromium：0.8 μ g/mL, cobalt：0.8μg/mL；
The concentration of 19 kinds of elements described in the high concentration quality-control product solution is respectively：Zinc：600 μ g/mL, strontium：300μ
g/mL；Selenium：200μg/mL；Lithium：100 μ g/mL, aluminium：100 μ g/mL, molybdenum：80 μ g/mL, arsenic：80 μ g/mL, iron：40 μ g/mL, barium：
40 μ g/mL, copper：20 μ g/mL, lead：8 μ g/mL, mercury：8 μ g/mL, nickel：8 μ g/mL, antimony：4 μ g/mL, manganese：4 μ g/mL, cadmium：4μg/
ML, thallium：2 μ g/mL, chromium：2 μ g/mL, cobalt：2μg/mL.
In wherein some embodiments, the standard solution includes the concentration of 9 standard curves, wherein 19 kinds of members
Element concentration be respectively：Zinc：5μg/mL、10μg/mL、20μg/mL、40μg/mL、125μg/mL、250μg/mL、500μg/mL、
1000μg/mL、2000μg/mL；
Strontium：2.5μg/mL、5μg/mL、10μg/mL、20μg/mL、62.5μg/mL、125μg/mL、250μg/mL、500μg/
mL、 1000μg/mL；
Selenium：1.5μg/mL、3μg/mL、6μg/mL、12μg/mL、37.5μg/mL、75μg/mL、150μg/mL、300μg/
mL、600μg/mL；
Lithium：1.5μg/mL、3μg/mL、6μg/mL、12μg/mL、37.5μg/mL、75μg/mL、150μg/mL、300μg/
mL、 600μg/mL；
Aluminium：1.5μg/mL、3μg/mL、6μg/mL、12μg/mL、37.5μg/mL、75μg/mL、150μg/mL、300μg/
mL、 600μg/mL；
Molybdenum：0.75μg/mL、1.5μg/mL、3μg/mL、6μg/mL、18.75μg/mL、37.5μg/mL、75μg/mL、150μ
g/mL、 300μg/mL；
Arsenic：0.75μg/mL、1.5μg/mL、3μg/mL、6μg/mL、18.75μg/mL、37.5μg/mL、75μg/mL、150μ
g/mL、 300μg/mL；
Iron：0.75μg/mL、1.5μg/mL、3μg/mL、6μg/mL、18.75μg/mL、37.5μg/mL、75μg/mL、150μ
g/mL、 300μg/mL；
Barium：0.5μg/mL、1μg/mL、2μg/mL、4μg/mL、12.5μg/mL、25μg/mL、50μg/mL、100μg/mL、
200μg/mL；
Copper：0.25μg/mL、0.5μg/mL、1μg/mL、2μg/mL、6.25μg/mL、12.5μg/mL、25μg/mL、50μg/
mL、 100μg/mL；
Lead：0.1μg/mL、0.2μg/mL、0.4μg/mL、0.8μg/mL、2.5μg/mL、5μg/mL、10μg/mL、20μg/
mL、 40μg/mL；
Mercury：0.1μg/mL、0.2μg/mL、0.4μg/mL、0.8μg/mL、2.5μg/mL、5μg/mL、10μg/mL、20μg/
mL、 40μg/mL；
Nickel：0.1μg/mL、0.2μg/mL、0.4μg/mL、0.8μg/mL、2.5μg/mL、5μg/mL、10μg/mL、20μg/
mL、 40μg/mL；
Antimony：0.05μg/mL、0.1μg/mL、0.2μg/mL、0.4μg/mL、1.25μg/mL、2.5μg/mL、5μg/mL、10μ
g/mL、 20μg/mL；
Manganese：0.05μg/mL、0.1μg/mL、0.2μg/mL、0.4μg/mL、1.25μg/mL、2.5μg/mL、5μg/mL、10μ
g/mL、 20μg/mL；
Cadmium：0.05μg/mL、0.1μg/mL、0.2μg/mL、0.4μg/mL、1.25μg/mL、2.5μg/mL、5μg/mL、10μ
g/mL、 20μg/mL；
Thallium：0.025μg/mL、0.05μg/mL、0.1μg/mL、0.2μg/mL、0.625μg/mL、1.25μg/mL、2.5μg/
mL、 5μg/mL、10μg/mL；
Chromium：0.025μg/mL、0.05μg/mL、0.1μg/mL、0.2μg/mL、0.625μg/mL、1.25μg/mL、2.5μg/
mL、 5μg/mL、10μg/mL；
Cobalt：0.025μg/mL、0.05μg/mL、0.1μg/mL、0.2μg/mL、0.625μg/mL、1.25μg/mL、2.5μg/
mL、 5μg/mL、10μg/mL。
Above-mentioned detection method can detect 19 kinds of elements in human body simultaneously, and analyze speed is fast, and detection flux is high；Sampling side
Method is simple, sample is easily stored, convenient transportation, even if situations such as sample is rotten will not occur in long-distance transport；Pre-treatment is simple,
High sensitivity, high specificity, interference are small, being capable of the relatively low element of accurate quantitative analysis concentration.
Specific embodiment
For the ease of understanding the present invention, the present invention will be described more fully below.But the present invention can be with perhaps
Mostly different form is realized, however it is not limited to embodiment described herein.On the contrary, the purpose for providing these embodiments is to make
To the understanding more thorough and comprehensive of the disclosure.
Unless otherwise defined, all of technologies and scientific terms used here by the article is with belonging to technical field of the invention
The normally understood meaning of technical staff is identical.Term used in the description of the invention herein is intended merely to description tool
The purpose of the embodiment of body, it is not intended that in the limitation present invention.Term as used herein "and/or" includes one or more phases
The arbitrary and all combination of the Listed Items of pass.
Embodiment 1
The detection method of 19 kinds of elements of the present embodiment, includes the following steps：
First, main reagent and instrument
U.S. Agilent Technologies ICP-MS (7900), IKA VOTEX3 whirlpool oscillators, Germany
Eppendorf pipettors and centrifuge tube, Vitlab volumetric flasks and jug, A Μ W220D electronic balances etc..
Μ P-S level isopropanols, analysis level-one water as defined in water is GB/T6682, resistivity >=18.2M Ω .cm, LC-
MS/MS grades of water (Fisher Chemical W6-4), top pure grade nitric acid, 1000mg/L goldstandard solution, Sigma
The Triton X-100Bioxtra of Aldrich, argon gas, liquid argon>99.999%.
2nd, the configuration of solution
2.1 dilutions (2mg/L gold marks, 0.1%Triton X-100,0.1% nitric acid)
200mL Fisher deionized waters are taken in 250mL volumetric flasks, add in 65% concentrated nitric acid 385 μ L, 500 μ L
The goldstandard solution of 1000mg/L, 250 μ L Triton X-100 solution, is shaken up so that nitric acid with Fisher deionized water constant volumes
Content be 0.1%.
2.2 standard solution
2.2.1 19 Heavy Metallic Elements Standard Stock solutions TS1
10mL Fisher deionized waters are taken in 100mL volumetric flasks, add in 65% concentrated nitric acids of 3mL, add in golden standard liquid 300
μ L, it (is specially zinc 10mL, strontium 5mL, selenium 3mL, lithium 3mL, aluminium 3mL, molybdenum 1.5mL, arsenic 1.5mL, iron to be separately added into respective volume
1.5mL, barium 1mL, 500 μ L of copper, 200 μ L of lead, 200 μ L of mercury, 200 μ L of nickel, 100 μ L of antimony, 100 μ L of manganese, 100 μ L of cadmium, 50 μ L of thallium,
50 μ L of chromium, 50 μ L of cobalt) a concentration of 1000mg/L element standard, with deionized water constant volume, shake up, be transferred to 100mL slots
In bottle, deposit in spare under normal temperature condition.Each ion concentration is respectively zinc 100ppm, strontium 50ppm, selenium 30ppm, lithium at this time
30ppm, aluminium 30ppm, molybdenum 15ppm, arsenic 15ppm, iron 15ppm, barium 10ppm, copper 5ppm, lead 2ppm, mercury 2ppm, nickel 2ppm, antimony
1ppm, manganese 1ppm, cadmium 1ppm, thallium 0.5ppm, chromium 0.5ppm, cobalt 0.5ppm.
2.2.2 standard curve TS2
10mL Fisher deionized waters are taken in 100mL volumetric flasks, 20 μ L gold standard liquids of addition, the concentrated nitric acid of 3mL 65%,
TS1 storing solution 1mL add Fisher deionized water constant volumes, shake up, and deposit in spare under room temperature.Each ion concentration point at this time
Not Wei zinc 1000ppb, strontium 500ppb, selenium 300ppb, lithium 300ppb, aluminium 300ppb, molybdenum 150ppb, arsenic 150ppb, iron 150ppb,
Barium 100ppb, copper 50ppb, lead 20ppb, mercury 20ppb, nickel 20ppb, antimony 10ppb, manganese 10ppb, cadmium 10ppb, thallium 5ppb, chromium
5ppb, cobalt 5ppb.
2.2.3 the configuration of Working calibration curve
The configuration of Working calibration curve such as the following table 1,
The configuration of 1. Working calibration curve of table
2. Working calibration curve concentration of table
2.3 inner mark solution
2.3.1 19 internal standard working solutions of element
10mL Fisher deionized waters are added in the volumetric flask of 100mL, add in the concentrated nitric acid of 3mL 65%, are added in
15mL Μ P-S level isopropanols, then the rhodium Rh storing solutions of the gallium Ga storing solutions (1000mg/L) of 200 μ L, 200 μ L are drawn respectively
(1000mg/L) mixing adds Fisher deionized water constant volumes, shakes up, spare under room temperature.
The preparation of 2.4 Quality Controls
2.4.1 19 Quality Control stock solution TQS1 of element
It first prepares 100mL and contains only 2% nitric acid dilution of 3mg/L gold target：10mL Fisher deionized waters are added to
In the volumetric flask of 100mL, the concentrated nitric acid of 3mL 65% is added in, 300 μ L of golden standard liquid is added in, is settled to Fisher deionized waters
Scale shakes up.
9 2mL centrifuge tubes are taken, 900 μ L is respectively added in and contains only golden target nitric acid dilution, then respectively add in 100 μ L marks
Quasi- product lead (1000mg/L), mercury (1000mg/L), nickel (1000mg/L), antimony (1000mg/L), manganese (1000mg/L), cadmium
(1000mg/L), thallium (1000mg/L), chromium (1000mg/L) and cobalt (1000mg/L) carry out standard items 10 times of dilutions to reach
Purpose, each ion concentration is respectively lead 100ppm, mercury 100ppm, nickel 100ppm, antimony 100ppm, manganese 100ppm, cadmium at this time
100ppm, thallium 100ppm, chromium 100ppm and cobalt 100ppm.
2mL Fisher deionized waters are taken in 10mL volumetric flasks, add in 300 μ L, 65% concentrated nitric acids, add in 30 μ of golden standard liquid
L adds in standard items zinc (1000mg/L), strontium (1000mg/L), selenium (1000mg/L), lithium (1000mg/L), aluminium (1000 mg/
L), molybdenum (1000mg/L), arsenic (1000mg/L), iron (1000mg/L), barium (1000mg/L), copper (1000mg/L) and dilute
Release 10 times standard items lead (100mg/L), mercury (100mg/L), nickel (100mg/L), antimony (100mg/L), manganese (100mg/L), cadmium
The amount of (100mg/L), thallium (100mg/L), chromium (100mg/L) and cobalt (100mg/L) is respectively 600 μ L, 300 μ L, 200 μ L, 100
μL、100μL、80μL、80μL、40μL、40μL、20μL、80μL、80μL、 80μL、40μL、40μL、40μL、20μL、20μL、20
μ L with Fisher deionized water constant volumes, shake up, and deposit in spare under normal temperature condition.At this time each ion concentration be respectively 60ppm,
30ppm、20ppm、10ppm、10ppm、8ppm、 8ppm、4ppm、4ppm、2ppm、0.8ppm、0.8ppm、0.8ppm、
0.4ppm、0.4ppm、0.4ppm、 0.2ppm、0.2ppm、0.2ppm。
2.4.2 work Quality Control configuration
The work Quality Control configuration method of table 3
The work Quality Control concentration of table 4
3rd, sample collection
The urine of 3.1 acquisition random time points preserves sample in the special urinary catheters of 15mL under refrigerated condition.
3.2 sample pre-treatments
Urine specimen is equilibrated into room temperature, pipettes 800 μ L samples in 10ml round bottom centrifuge tubes, adds in 3.2mL dilutions,
Concussion 1min is shaken up at a high speed to get sample to be tested.
It should be understood that urine sample can also be directly over diluted, (the two volume ratio is 1:4), after mixing
Obtain sample to be tested.
Dilution be 2mg/L gold marks, 0.1%Triton X-100,0.1% nitric acid：Take 200mL Fisher deionized waters
In 250mL volumetric flasks, 65% 385 μ L of concentrated nitric acid, the goldstandard solution of 500 μ L 1000mg/L, 250 μ L are added in
Triton X-100 solution is shaken up with Fisher deionized water constant volumes so that the content of nitric acid is 0.1%.
The sample to be tested is detected using inductivity coupled plasma mass spectrometry, method is simple, and the time, short flux was high, sensitive
Degree and specificity are high, and accuracy and precision are good.By detection obtain in urine the concentration of various elements and with creatinine in urine
It carries out than being worth to final result.
4th, it detects
4.1 introduce systematic parameter (MS conditions)
Atomizer temperature：2℃
Table 5 introduces systematic parameter
4.2 source parameters (ion so μ rce conditions)
6 acquisition parameter of table
7 mass spectrometry parameters of table
4.3 element mass numbers
Table 8
4.4 ICP-MS programs
Urine sample obtains sample to be tested after 5 times of dilution, will by peristaltic pump by the sample handled well by peristaltic pump
Fluid sample and internal standard solution are loaded onto atomizer automatically simultaneously.Using argon gas as carrier gas by the solution atomization of introducing, sample is molten
Liquid passes through bilateral type spray chamber after forming small droplet, in the spray chamber, by the shock touched with atomization locular wall, removes larger
Sample drop, the sample aerosol of formation are directly entered in ICP-MS quarter bend central-injection pipelines.Pass through high temperature in quarter bend end
Plasma (4500-6500K) by sample aerosol moment precipitation and ionization, charged ion under voltage and vacuum action,
After being focused on into ion lens, into collision cell, collided here with intert-gas atoms, polyatomic molecule is fragmented into respectively
Monatomic ion, subsequently into triple level four bars mass analyzers.In level four bars mass analyzer, by changing voltage,
The ion of the specified mass-to-charge ratio of screening.After quadrupole rod, the signal of ion is measured using electron multiplier.Electron multiplier (EM)
There are many dynodes.When ion enters EM, the first dynode of ionic bombardment simultaneously generates electronics rain.Then it hits next
Electron multiplication electrode generates more electronics.After a series of dynode, the electronics of multiplication is detected as e-mail
Number.Obtained data are transferred to computer, and computer counts collected number of ions to get to each population of ions number
According to.
Zinc, strontium, selenium, lithium, aluminium, molybdenum, arsenic, iron, barium, copper, lead, mercury, nickel, calcium halophosphate activated by antimony andmanganese, cadmium, thallium, chromium and cobalt in reference substance and
The ratio that its interior target counts be used to build internal standard standard curve, and then the curve be used to calculate sample or quality control substance
In zinc, strontium, selenium, lithium, aluminium, molybdenum, arsenic, iron, barium, copper, lead, mercury, nickel, calcium halophosphate activated by antimony andmanganese, cadmium, thallium, chromium and the respective concentration of cobalt.
2 methodology validation of embodiment
The present embodiment carries out methodology validation to the detection method of 19 kinds of elements in embodiment 1.
1. precision
Withinrun precision：Basic, normal, high three horizontal Quality Control samples is taken to carry out withinrun precision experiment；Each concentration level
Sample parallel processing 20, each sample introduction 1 time；I.e. each concentration level obtains 20 groups of data, calculate testing result average value and
Relative standard deviation (RSD).
Betweenrun precision：Basic, normal, high three horizontal quality-control product prepared in advance is taken, carries out betweenrun precision experiment；Often
The sample of a level does 4 parallel, METHOD FOR CONTINUOUS DETERMINATIONs 5 days respectively；I.e. each concentration level obtains 20 groups of data, calculates testing result
Average value and relative standard deviation (RSD).
Precision verification result：Precision Experiment verification result is as shown in table 9, and withinrun precision is small with betweenrun precision
In 15%, meet testing requirements.
The Precision test result of 9 19 kinds of elements of table
2. sensitivity for analysis and the range of linearity
2.1 method quantitative limits and the range of linearity
A. bare substrate is prepared；
B. configuration standard curve；
C. the sample parallel processing of each concentration 6 detects primary (n=6) respectively；
D. the average value, RSD and the rate of recovery of each concentration samples are calculated；
E. method quantitative limit calibrates standard really：RSD is less than 20% and the rate of recovery is minimum dense in the range of 85%-115%
Degree point is considered as quantitative limit concentration, i.e. LOQ；
F. the range of linearity calibrates standard really：RSD is less than 20%, and the rate of recovery is in the range of 85%-115%, and with theoretical dense
The regression curve r that degree ratio is depicted as with actual signal response peak area ratio>0.98, that is, meet the requirement of the range of linearity；
G. confirmatory experiment data are as shown in following table 10-29.
10 bronze medal quantitative limit experimental data of table
11 zinc quantitative limit experimental data of table
12 strontium quantitative limit experimental data of table
13 molybdenum quantitative limit experimental data of table
14 antimony quantitative limit experimental data of table
15 barium quantitative limit experimental data of table
16 mercury quantitative limit experimental data of table
17 thallium quantitative limit experimental data of table
18 lithium quantitative limit experimental data of table
19 aluminium quantitative limit experimental data of table
20 chromium quantitative limit experimental data of table
21 manganese quantitative limit experimental data of table
22 iron quantitative limit experimental data of table
23 nickel quantitative limit experimental data of table
24 cobalt quantitative limit experimental data of table
25 arsenic quantitative limit experimental data of table
26 selenium quantitative limit experimental data of table
27 cadmium quantitative limit experimental data of table
28 lead quantitative limit experimental data of table
29 equation of linear regression of table and method quantitative limit
2.2 method detection limits
Collect patient's sample of the concentration near quantitative limit LOQ；Parallel processing this patient's sample 12, detects 2 respectively
It is secondary；I.e. each sample obtains 24 groups of data, calculates average value, standard deviation (SD), the coefficient of variation (CV) and the method detection of each sample
It limits (LOD)；
Experimental result is as shown in table 30, and the LOD of copper is 0.156 μ g/L, and the LOD of zinc is 1.155 μ g/L, and the LOD of strontium is
0.217 μ g/L, the LOD of molybdenum is 0.268 μ g/L, and the LOD of antimony is 0.015 μ g/L, and the LOD of barium is 0.177 μ g/L, and the LOD of mercury is
0.080 μ g/L, the LOD of thallium is 0.015 μ g/L, and the LOD of lithium is 1.150 μ g/L, and the LOD of aluminium is 1.207 μ g/L, and the LOD of chromium is
0.053 μ g/L, the LOD of manganese is 0.057 μ g/L, and the LOD of iron is 0.513 μ g/L, and the LOD of nickel is 0.077 μ g/L, and the LOD of cobalt is
0.014 μ g/L, the LOD of arsenic is 0.232 μ g/L, and the LOD of selenium is 0.814 μ g/L, and the LOD of cadmium is 0.054 μ g/L, and the LOD of lead is
0.073μg/L。
30 method detection limit result of the test of table
3. accuracy
Recovery test：Random collecting a batch pooled serum sample, measures base concentration, is separately added into basic, normal, high three water
The standard items of flat concentration, with same steps reprocessing and measurement result；Every group of parallel processing 4, it is each to detect 2 times, it calculates
The mark-on sample result rate of recovery.As shown in table 31, the recovery of standard addition of 19 kinds of elements is full between 88.57%~116.10%
The requirement of foot 100% ± 20%, it is believed that method is accurate.
The recovery of standard addition result of the test of 31 19 kinds of elements of table
Each technical characteristic of embodiment described above can be combined arbitrarily, to make description succinct, not to above-mentioned reality
It applies all possible combination of each technical characteristic in example to be all described, as long as however, the combination of these technical characteristics is not deposited
In contradiction, it is all considered to be the range of this specification record.
Embodiment described above only expresses the several embodiments of the present invention, and description is more specific and detailed, but simultaneously
It cannot therefore be construed as limiting the scope of the patent.It should be pointed out that those of ordinary skill in the art are come
It says, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to the protection of the present invention
Range.Therefore, the protection domain of patent of the present invention should be determined by the appended claims.
Claims (7)
1. detection method that is a kind of while detecting 19 kinds of elements of human body, which is characterized in that include the following steps：
19 kinds of elements are respectively：Zinc, strontium, selenium, lithium, aluminium, molybdenum, arsenic, iron, barium, copper, lead, mercury, nickel, calcium halophosphate activated by antimony andmanganese, cadmium, thallium,
Chromium and cobalt；
(1) solution is prepared
Dilution：Gold containing 1.5-2.5mg/L is marked with and the 0.1-2wt% aqueous solution of nitric acid of 0.05-0.2wt% Triton X-100s；
Standard solution：It is formulated by adding in 19 kinds of element standards in the dilution, wherein 19 kinds of members
Element concentration be respectively：Zinc：5-2000 μ g/mL, strontium：2.5-1000μg/mL；Selenium 1.5-600 μ g/mL；Lithium：1.5-600μg/
ML, aluminium：1.5-600 μ g/mL, molybdenum：0.75-300 μ g/mL, arsenic：0.75-300 μ g/mL, iron：0.75-300 μ g/mL, barium：0.5-
200 μ g/mL, copper：0.25-100 μ g/mL, lead：0.1-40 μ g/mL, mercury：0.1-40 μ g/mL, nickel：0.1-40 μ g/mL, antimony：
0.05-20 μ g/mL, manganese：0.05-20 μ g/mL, cadmium：0.05-20 μ g/mL, thallium：0.025-10 μ g/mL, chromium：0.025-10μg/
ML, cobalt：0.025-10μg/mL；
Inner mark solution：It is formulated by adding in internal standard element standard items in the dilution, the internal standard element is gallium and rhodium；
(2) sample to be tested
Acquire sample：Collect random time point urine；
Sample pre-treatments：The urine is through the diluted to get the sample to be tested；
(3) it detects
Inner mark solution is added in the standard solution, is detected using Plasma-Mass Spectroscopy ICP-MS, builds standard curve；Wherein
Mass Spectrometry Conditions are：
Add in inner mark solution in the sample to be tested, using Plasma-Mass Spectroscopy ICP-MS detect to get.
2. detection method that is according to claim 1 while detecting 19 kinds of elements of human body, which is characterized in that the collection
The step of sample, further includes：The urine of random time point is acquired, filter paper is taken to infiltrate urine, dries, obtains dry nappy sample.
3. detection method that is according to claim 2 while detecting 19 kinds of elements of human body, which is characterized in that the sample
The step of pre-treatment, further includes：The dry nappy sample described in distillation water wash, centrifuges to obtain the sample to be tested.
4. detect the detection method of 19 kinds of elements of human body simultaneously according to claim 1-3 any one of them, which is characterized in that
Quality-control product solution is further included in the solution preparation steps：
It is formulated by adding in 19 kinds of element standards in the dilution, the quality-control product solution includes low concentration matter
Three kinds of control product solution, middle concentration quality-control product solution and high concentration quality-control product solution concentration；
The concentration of 19 kinds of elements described in the low concentration quality-control product solution is respectively：Zinc：120 μ g/mL, strontium：60μg/mL；
Selenium：40μg/mL；Lithium：20 μ g/mL, aluminium：20 μ g/mL, molybdenum：16 μ g/mL, arsenic：16 μ g/mL, 8 μ g/mL of iron, barium：8 μ g/mL, copper：
4 μ g/mL, lead：1.6 μ g/mL, mercury：1.6 μ g/mL, nickel：1.6 μ g/mL, antimony：0.8 μ g/mL, manganese：0.8 μ g/mL, cadmium：0.8μg/
ML, thallium：0.4 μ g/mL, chromium：0.4 μ g/mL, cobalt：0.4μg/mL；
The concentration of 19 kinds of elements described in the middle concentration quality-control product solution is respectively：Zinc：240 μ g/mL, strontium：120μg/mL；
Selenium：80μg/mL；Lithium：40 μ g/mL, aluminium：40 μ g/mL, molybdenum：32 μ g/mL, arsenic：32 μ g/mL, iron：16 μ g/mL, barium：16 μ g/mL,
Copper：8 μ g/mL, lead：3.2 μ g/mL, mercury：3.2 μ g/mL, nickel：3.2 μ g/mL, antimony：1.6 μ g/mL, manganese：1.6 μ g/mL, cadmium：1.6μ
G/mL, thallium：0.8 μ g/mL, chromium：0.8 μ g/mL, cobalt：0.8μg/mL；
The concentration of 19 kinds of elements described in the high concentration quality-control product solution is respectively：Zinc：600 μ g/mL, strontium：300μg/mL；
Selenium：200μg/mL；Lithium：100 μ g/mL, aluminium：100 μ g/mL, molybdenum：80 μ g/mL, arsenic：80 μ g/mL, iron：40 μ g/mL, barium：40μg/
ML, copper：20 μ g/mL, lead：8 μ g/mL, mercury：8 μ g/mL, nickel：8 μ g/mL, antimony：4 μ g/mL, manganese：4 μ g/mL, cadmium：4 μ g/mL, thallium：
2 μ g/mL, chromium：2 μ g/mL, cobalt：2μg/mL.
5. kit that is a kind of while detecting 19 kinds of elements of human body, which is characterized in that including dilution, standard solution and interior
Mark solution；
Gold containing 1.5-2.5mg/L is marked with and the 0.1-2wt% aqueous solution of nitric acid of 0.05-0.2wt% Triton X-100s；
The standard solution：It is formulated by adding in 19 kinds of element standards in the dilution, wherein described 19
The concentration of kind of element is respectively：Zinc：5-2000 μ g/mL, strontium：2.5-1000μg/mL；Selenium 1.5-600 μ g/mL；Lithium：1.5-600μ
G/mL, aluminium：1.5-600 μ g/mL, molybdenum：0.75-300 μ g/mL, arsenic：0.75-300 μ g/mL, iron：0.75-300 μ g/mL, barium：
0.5-200 μ g/mL, copper：0.25-100 μ g/mL, lead：0.1-40 μ g/mL, mercury：0.1-40 μ g/mL, nickel：0.1-40 μ g/mL,
Antimony：0.05-20 μ g/mL, manganese：0.05-20 μ g/mL, cadmium：0.05-20 μ g/mL, thallium：0.025-10 μ g/mL, chromium：0.025-10μ
G/mL, cobalt：0.025-10μg/mL；
The inner mark solution：Be formulated by adding in internal standard element standard items in the dilution, the internal standard element for gallium and
Rhodium, a concentration of 1000 μ g/mL of gallium, a concentration of 1000 μ g/mL of rhodium.
6. kit that is according to claim 5 while detecting 19 kinds of elements of human body, which is characterized in that further include Quality Control
Product solution：
It is formulated by adding in 19 kinds of element standards in the dilution, the quality-control product solution includes low concentration matter
Three kinds of control product solution, middle concentration quality-control product solution and high concentration quality-control product solution concentration；
The concentration of 19 kinds of elements described in the low concentration quality-control product solution is respectively：Zinc：120 μ g/mL, strontium：60μg/mL；
Selenium：40μg/mL；Lithium：20 μ g/mL, aluminium：20 μ g/mL, molybdenum：16 μ g/mL, arsenic：16 μ g/mL, 8 μ g/mL of iron, barium：8 μ g/mL, copper：
4 μ g/mL, lead：1.6 μ g/mL, mercury：1.6 μ g/mL, nickel：1.6 μ g/mL, antimony：0.8 μ g/mL, manganese：0.8 μ g/mL, cadmium：0.8μg/
ML, thallium：0.4 μ g/mL, chromium：0.4 μ g/mL, cobalt：0.4μg/mL；
The concentration of 19 kinds of elements described in the middle concentration quality-control product solution is respectively：Zinc：240 μ g/mL, strontium：120μg/mL；
Selenium：80μg/mL；Lithium：40 μ g/mL, aluminium：40 μ g/mL, molybdenum：32 μ g/mL, arsenic：32 μ g/mL, iron：16 μ g/mL, barium：16 μ g/mL,
Copper：8 μ g/mL, lead：3.2 μ g/mL, mercury：3.2 μ g/mL, nickel：3.2 μ g/mL, antimony：1.6 μ g/mL, manganese：1.6 μ g/mL, cadmium：1.6μ
G/mL, thallium：0.8 μ g/mL, chromium：0.8 μ g/mL, cobalt：0.8μg/mL；
The concentration of 19 kinds of elements described in the high concentration quality-control product solution is respectively：Zinc：600 μ g/mL, strontium：300μg/mL；
Selenium：200μg/mL；Lithium：100 μ g/mL, aluminium：100 μ g/mL, molybdenum：80 μ g/mL, arsenic：80 μ g/mL, iron：40 μ g/mL, barium：40μg/
ML, copper：20 μ g/mL, lead：8 μ g/mL, mercury：8 μ g/mL, nickel：8 μ g/mL, antimony：4 μ g/mL, manganese：4 μ g/mL, cadmium：4 μ g/mL, thallium：
2 μ g/mL, chromium：2 μ g/mL, cobalt：2μg/mL.
7. kit that is according to claim 5 or 6 while detecting 19 kinds of elements of human body, which is characterized in that the mark
Quasi- solution includes the concentration of 9 standard curves, wherein the concentration of 19 kinds of elements is respectively：Zinc：5μg/mL、10μg/mL、
20μg/mL、40μg/mL、125μg/mL、250μg/mL、500μg/mL、1000μg/mL、2000μg/mL；
Strontium：2.5μg/mL、5μg/mL、10μg/mL、20μg/mL、62.5μg/mL、125μg/mL、250μg/mL、500μg/mL、
1000μg/mL；
Selenium：1.5μg/mL、3μg/mL、6μg/mL、12μg/mL、37.5μg/mL、75μg/mL、150μg/mL、300μg/mL、600
μg/mL；
Lithium：1.5μg/mL、3μg/mL、6μg/mL、12μg/mL、37.5μg/mL、75μg/mL、150μg/mL、300μg/mL、600
μg/mL；
Aluminium：1.5μg/mL、3μg/mL、6μg/mL、12μg/mL、37.5μg/mL、75μg/mL、150μg/mL、300μg/mL、600
μg/mL；
Molybdenum：0.75μg/mL、1.5μg/mL、3μg/mL、6μg/mL、18.75μg/mL、37.5μg/mL、75μg/mL、150μg/
mL、300μg/mL；
Arsenic：0.75μg/mL、1.5μg/mL、3μg/mL、6μg/mL、18.75μg/mL、37.5μg/mL、75μg/mL、150μg/
mL、300μg/mL；
Iron：0.75μg/mL、1.5μg/mL、3μg/mL、6μg/mL、18.75μg/mL、37.5μg/mL、75μg/mL、150μg/
mL、300μg/mL；
Barium：0.5μg/mL、1μg/mL、2μg/mL、4μg/mL、12.5μg/mL、25μg/mL、50μg/mL、100μg/mL、200μ
g/mL；
Copper：0.25μg/mL、0.5μg/mL、1μg/mL、2μg/mL、6.25μg/mL、12.5μg/mL、25μg/mL、50μg/mL、
100μg/mL；
Lead：0.1μg/mL、0.2μg/mL、0.4μg/mL、0.8μg/mL、2.5μg/mL、5μg/mL、10μg/mL、20μg/mL、40
μg/mL；
Mercury：0.1μg/mL、0.2μg/mL、0.4μg/mL、0.8μg/mL、2.5μg/mL、5μg/mL、10μg/mL、20μg/mL、40
μg/mL；
Nickel：0.1μg/mL、0.2μg/mL、0.4μg/mL、0.8μg/mL、2.5μg/mL、5μg/mL、10μg/mL、20μg/mL、40
μg/mL；
Antimony：0.05μg/mL、0.1μg/mL、0.2μg/mL、0.4μg/mL、1.25μg/mL、2.5μg/mL、5μg/mL、10μg/
mL、20μg/mL；
Manganese：0.05μg/mL、0.1μg/mL、0.2μg/mL、0.4μg/mL、1.25μg/mL、2.5μg/mL、5μg/mL、10μg/
mL、20μg/mL；
Cadmium：0.05μg/mL、0.1μg/mL、0.2μg/mL、0.4μg/mL、1.25μg/mL、2.5μg/mL、5μg/mL、10μg/
mL、20μg/mL；
Thallium：0.025μg/mL、0.05μg/mL、0.1μg/mL、0.2μg/mL、0.625μg/mL、1.25μg/mL、2.5μg/mL、5
μg/mL、10μg/mL；
Chromium：0.025μg/mL、0.05μg/mL、0.1μg/mL、0.2μg/mL、0.625μg/mL、1.25μg/mL、2.5μg/mL、5
μg/mL、10μg/mL；
Cobalt：0.025μg/mL、0.05μg/mL、0.1μg/mL、0.2μg/mL、0.625μg/mL、1.25μg/mL、2.5μg/mL、5
μg/mL、10μg/mL。
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