CN103063732B - The assay method of chromium, nickel, arsenic, selenium, cadmium, mercury, lead content in a kind of urine - Google Patents
The assay method of chromium, nickel, arsenic, selenium, cadmium, mercury, lead content in a kind of urine Download PDFInfo
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Abstract
An assay method for chromium, nickel, arsenic, selenium, cadmium, mercury, lead content in urine, its test process is whole urine of collection smoker excretion in 24 hours and is stored in preservation at-80 DEG C in polypropylene bucket.Urine, after micro-wave digestion process, in acid condition, is carrier gas with argon gas, measures with inductivity coupled plasma mass spectrometry, and more quantitative with standard series.The present invention have measurement result accurately, sensitivity and the advantage such as reproducible, and operating process is easy, quick.
Description
Technical field:
The invention belongs to the collection analysis detection technique field of urine, relate generally to trace element determination techniques field in urine, is chromium in a kind of employing inductively coupled plasma mass spectrometry Accurate Determining smoker twenty-four-hour urine liquid specifically, nickel, arsenic, selenium, cadmium, mercury, the method for lead content.
Background technology:
Heavy metal, as harmful element important in cigarette smoke, can enter human body along with main flume, cause certain harm to human body.In the process of smoking property of cigarette, the heavy metal element in pipe tobacco can be used as flue gas aerosol and is scattered in main flume and side-stream smoke when cigarette burning, and it enters after in body can accumulate in vivo.Reacted by Fenton, generate free radical, free radical can be combined with DNA, causes the oxidative damage of DNA.Heavy metal is considered to the major reason causing a lot of disease.
Wang little Yan etc. utilize inductivity coupled plasma mass spectrometry (ICP-MS), establish 7 method of Heavy Metallic Elements content of Cr, Mn, As, Se, Cd, Hg, Pb etc. in direct dilution metering blood of human body and urine.Abundant once waited with aas determination crowd's blood of being exposed to electronic waste processing environment and urinated in lead, cadmium, copper level, and the heavy metal level of analyzing and researching in subject and Correlative Influence Factors thereof.
Current research smoking there is not yet report to the impact of smoker's heavy metal of body level.Utilize atomic absorption spectrum can not detect trace heavy metal element in multiple smoker's urine fast and accurately simultaneously.For the impact of heavy metals in smoker's body that smoking causes, need the heavy metal element existed in eliminating diet and environment on the impact of smoker's heavy metal of body.Therefore the interference getting rid of diet and environment in urine capture process is very crucial.Described in summary, be necessary the Simultaneously test method setting up detection device for multi metallic elements content in a kind of urine.Be used for getting rid of the interference for smoker's heavy metal of body element of diet and environment, and detect detection device for multi metallic elements fast and accurately simultaneously.
Summary of the invention:
Object of the present invention is intended to set up a kind of Inductively coupled plasma-mass spectrometry that adopts and measures seven heavy metal species (chromium in smoker's urine, nickel, arsenic, selenium, cadmium, mercury and lead) method of content, the method can carry out Accurate Determining to seven heavy metal species content increases in smoker's body simultaneously, and measurement result accurately, to measure interference few.
The object of the invention is to be achieved through the following technical solutions: in mensuration smoker urine of the present invention, the assay method of seven heavy metal species content comprises following processing step:
A, urine capture, be divided into two groups of (smokings by volunteer; Non-smoking), gather the whole urine of volunteer in continuous 24 hours.
B, sample preparation, in polypropylene bucket, urine measures 10mL, inserts in micro-wave diminishing pot, then puts into microwave dissolver and carries out micro-wave digestion.
The micro-wave digestion heating schedule adopted is: intensification 20min to 95 DEG C, keeps 8min; Intensification 15min to 125 DEG C, keeps 8min; Intensification 20min to 165 DEG C, keeps 20min; Lower the temperature 20min to 20 DEG C, keep 20min; Be warming up to 190 DEG C, keep 10min.
The preparation of c, seven heavy metal species standard solution:
Chromium, nickel, arsenic, selenium, cadmium, the preparation of plumbous hybrid standard working solution: the hybrid standard product 0.500g accurately being pipetted 10 μ g/mL by electronic balance, use 5%(volume fraction) salpeter solution, use balance be accurately settled to 50.000g, concentration 100.000 μ g/L after constant volume.With balance pipette respectively 100.000 μ g/L mixed standard solution 0,0.250,0.500,1.000,2.000,5.000,10.000g, with 5%(V/V) dust technology dilution, use balance to be settled to 50.000,50.000,50.000,50.000,40.000,50.000,50.000g, shake up, join to obtain the serial mixed standard solution of 0,0.500,1.000,2.000,5.000,10.000,20.000 μ g/L respectively.
Preparation is containing the series standard solution of mercury: the mercury standard items 0.500g accurately being pipetted 10 μ g/mL by electronic balance, uses 5%(volume fraction) salpeter solution be settled to 50.000g, use concentration 100.000 μ g/L after balance constant volume.Pipette respectively the mercury standard solution 0 of 100.000 μ g/L, 0.250,0.500,1.000,2.000g, with 5%(V/V) dust technology dilution, use balance to be settled to 50.000,50.000,50.000,50.000,40.000g.Join to obtain the mercury standard solution of 0,0.500,1.000,2.000,5.000 μ g/L respectively.
D, sample determination, inductivity coupled plasma mass spectrometry measures, draw the heavy metal standard solution of the variable concentrations prepared, using argon gas as carrier, at high temperature the charged steam of standard solution with heavy metal element is blown into inductivity coupled plasma mass spectrometry to measure, be that 52 places record chromium in mass-to-charge ratio, 60 places record nickel, 75 places record arsenic, 82 places record selenium, 111 places record cadmium, 202 places record mercury, 208 places record lead, and use scandium, germanium, bismuth, indium is that interior mark corrects instrument, get the sample solution after micro-wave digestion to measure, the signal respective strengths and the concentration known standard solution that record seven Heavy Metallic Elements compare, the content of seven Heavy Metallic Elements in sample can be tried to achieve.
Carrier gas in sample determination process (argon gas) flow is 1.16L/min, and carrier fluid (nitric acid) concentration of volume percent is 5%.
The concrete technological process of the present invention is described in further detail as follows:
1. urine capture
Volunteer is divided into two groups of (smokings; Non-smoking) gather the whole urine of volunteer in continuous 24 hours.
2. micro-wave digestion sample
Accurately measure the urine 10mL in polypropylene bucket, be placed in counteracting tank.First use 1mL30%(v/v) hydrogen peroxide, 2.5mL65%(v/v) HNO
3add counteracting tank, the program of clearing up set by employing table 1 carries out digestion process.After (liquid that counteracting tank loads at most) clears up end, be down to after room temperature until temperature and take out counteracting tank, then digestion solution is transferred in 50mL volumetric flask, with 5%(v/v) HNO
3cleaning counteracting tank, and use 5%(v/v) HNO
3be settled to 50mL, to be measured.Blank test is done in company with sample.
Table 1 micro-wave digestion program
| Stage | 1 | 2 | 3 | 4 | 5 |
| Run energy (W) | 1200 | 1200 | 1200 | 1200 | 1200 |
| Heating-up time (min) | 20 | 15 | 20 | 20 | 20 |
| Terminal temperature (DEG C) | 95 | 125 | 165 | 20 | 190 |
| Retention time (min) | 8 | 8 | 15 | 20 | 10 |
3. inductivity coupled plasma mass spectrometry measures
3.1 inductivity coupled plasma mass spectrometry parameters
The experiment condition of inductivity coupled plasma mass spectrometry is as shown in table 2:
Table 2 Clod Vapour Atomic Absorption spectrometer parameter
| Parameter | Condition |
| Measure element | Chromium, nickel, arsenic, selenium, cadmium, mercury, plumbous |
| Radio frequency intensity | 1330w 2 --> |
| Sample depth | 7.5mm |
| Flow rate of carrier gas | 1.16L/min |
| Pump speed | 0.10rps |
The preparation of 3.2 7 heavy metal species standard solution:
The preparation of six heavy metal species hybrid standard working solutions: the hybrid standard product 0.500g accurately being pipetted 10 μ g/mL by electronic balance, uses 5%(volume fraction) salpeter solution, use balance be accurately settled to 50.000g, concentration 100.000 μ g/L after constant volume.With balance pipette respectively 100.000 μ g/L mixed standard solution 0,0.250,0.500,1.000,2.000,5.000,10.000g, with 5%(V/V) dust technology dilution, use balance to be settled to 50.000,50.000,50.000,50.000,40.000,50.000,50.000g, shake up, join to obtain the serial mixed standard solution of 0,0.500,1.000,2.000,5.000,10.000,20.000 μ g/L respectively.
The preparation of mercury standard working solution: the mercury standard items 0.500g accurately being pipetted 10 μ g/mL by electronic balance, uses 5%(volume fraction) salpeter solution be settled to 50.000g, use concentration 100.000 μ g/L after balance constant volume.Pipette respectively the amalgamation standard solution 0 of 100.000 μ g/L, 0.250,0.500,1.000,2.000g, with 5%(V/V) dust technology dilution, use balance to be settled to 50.000,50.000,50.000,50.000,40.000g.Join to obtain the mercury standard solution of 0,0.500,1.000,2.000,5.000 μ g/L respectively.
The mensuration of 3.3 7 Heavy Metallic Elements content
Be carrier gas with argon gas, according to inductivity coupled plasma mass spectrometry: the condition of work of 7500A type measures series standard working solution, obtains calibration curve; Test result is represented with the mean value of 2 replicate determinations.Calculate regression equation and the related coefficient of typical curve.The sample cancelled after separating process measures, and records the signal respective strengths of seven Heavy Metallic Elements, substitutes into the content that regression equation can obtain seven Heavy Metallic Elements in sample.
The repeatability of 3.4 the inventive method, detection limit and recovery of standard addition
The standard solution getting 1.000 μ g/L carries out 2 replicate determinations, and relative standard deviation (RSD) is chromium: 4.12%, nickel: 1.57%, arsenic: 2.61%, selenium: 1.00%, cadmium: 3.17%, mercury: 0.44%, plumbous, and 1.81%., according to this method, blank solution is measured meanwhile, obtain its detection limit with 3 times of standard deviations as shown in table 3.
Table 3 detection limit and quantitative limit (μ gL
-1)
| Chromium | Nickel | Arsenic | Selenium | Cadmium | Mercury | Plumbous | |
| LOD | 0.0753 | 0.0815 | 0.0090 | 0.0401 | 0.0157 | 0.0006 | 0.0437 |
| LOQ | 0.2259 | 0.2445 | 0.0270 | 0.1230 | 0.0471 | 0.0018 | 0.1311 |
Adopt this method, carried out measuring to the content of heavy metal in urine by the condition after optimizing and investigated the recovery of method, as shown in table 4.
Seven heavy metal species assay and recovery of standard addition in table 4 urine
The present invention establishes the mensuration of seven Heavy Metallic Elements in urine and the method for accurate preparing standard solution, proposes the effective collect means to smoker's urine.And use Inductively coupled plasma-mass spectrometry to detect seven Heavy Metallic Elements fast and accurately simultaneously.The inventive method has following excellent results:
1. volunteer is divided into two groups of (smokings; Non-smoking), gather twenty-four-hour urine liquid.
2. adopt micro-wave digestion mode can reduce sample and strong acid use amount, improve and clear up ability, and sample element loss to be measured is few, to operating personnel endanger little, be easy to operate.Digestion solution clarification, transparent, not precipitation.
3. adopt the quantitative preparing standard solution of electronic balance, than use volumetric flask preparing standard solution, accuracy effectively improves 3 orders of magnitude.
When 4. adopting inductive coupling plasma mass spectrometry to measure the mercury in cigarette mainstream flue gas, detect the recovery high, measurement result is accurate.
5. this method has easy and simple to handle, quick, accurate, sensitivity and reproducible advantage.
Accompanying drawing explanation
Fig. 1 is urine sample of smoker collecting flowchart figure (this figure is as Figure of abstract) of the present invention.
Embodiment
The present invention is described further below in conjunction with example, but is not restriction the present invention.
Volunteer is divided into two groups of (smokings; Non-smoking), gather twenty-four-hour urine liquid.
After collection completes, sample preparation, accurately measures urine 10mL, inserts in micro-wave diminishing pot, then puts into microwave dissolver and carries out micro-wave digestion.(liquid that counteracting tank loads at most)
After clearing up end, be down to after room temperature until temperature and take out counteracting tank, after clearing up end, be down to after room temperature until temperature and take out counteracting tank, then digestion solution is transferred in 50mL volumetric flask, with 5%(v/v) HNO
3cleaning counteracting tank, and use 5%(v/v) HNO
3be settled to 50mL, to be measured.Blank test is done in company with sample.
Sample determination, inductivity coupled plasma mass spectrometry measures, draw the heavy metal standard working solution of the variable concentrations prepared, using argon gas as carrier, at high temperature will be blown into inductivity coupled plasma mass spectrometry with the charged steam of heavy metal atom sample and measure.Be that 52 places record chromium in mass-to-charge ratio, 60 places record nickel, and 75 places record arsenic, and 82 places record selenium, and 111 places record cadmium, and 202 places record mercury, and 208 places record lead.And using scandium, germanium, bismuth, indium is interior scalar quantity.The sample solution cancelled after solution measures, and the intensity and the standard items that record seven Heavy Metallic Elements contrast, and can try to achieve the content of seven Heavy Metallic Elements in sample.
Example 1:
1. instrument and reagent: inductively coupled plasma: Agilent company, 7500A type inductivity coupled plasma mass spectrometry; Mars5 type microwave dissolver (U.S. CE M company), joins the high-pressure sealed counteracting tank of PTFE; Electronic balance, CP225D type, sensibility reciprocal 0.0001g(SartoriusAG company); Argon gas: purity is 99.99%;
Ultrapure water.Mass percent is the red fuming nitric acid (RFNA) of 65%, mass percent be 30% hydrogen peroxide be chromatographically pure; Hybrid standard product and mercury standard items (10.0mg/L, Agilent company of the U.S.).
2. sample preparation: in polypropylene bucket, urine measures 10mL, inserts in micro-wave diminishing pot, then puts into microwave dissolver and carries out micro-wave digestion.(liquid that counteracting tank loads at most) uses 1mL30%(v/v successively) hydrogen peroxide, the red fuming nitric acid (RFNA) of 2.5mL65% is inserted in counteracting tank, then puts into microwave dissolver, and the program of clearing up set by employing table 1 carries out digestion process.After (due to the liquid that counteracting tank loads at most) clears up end, be down to after room temperature until temperature and take out counteracting tank, then digestion solution to be transferred in 50mL volumetric flask and constant volume is to be measured.
3. assay method:
With carrier gas (argon gas) flow 1.16L/min, open argon gas, recirculated water, inductivity coupled plasma mass spectrometry, open working software, by table 2, mass spectrometry parameters is set, absorption standard blank solution, prepare 0,0.500,1.000,2.000,5.000,10.000,20.000 μ g/L hybrid standard working solutions and the sample after clearing up enter mass spectrometer and measure.Record seven heavy metal species total amounts in twenty-four-hour urine liquid sample as follows:
| Chromium | Nickel | Arsenic | Selenium | Cadmium | Mercury | Plumbous |
| μg | μg | μg | μg | μg | μg | μg |
| 41.5872 | 3.1872 | 38.3901 | 24.1704 | 0.8406 | 0.8278 | 3.9976 |
Example 2:
As described in Example 1, select the smoking volunteer urine sample being numbered B, record twenty-four-hour urine liquid sample total metals as follows:
| Chromium | Nickel | Arsenic | Selenium | Cadmium | Mercury | Plumbous |
| μg | μg | μg | μg | μg | μg | μg |
| 21.1443 | 8.8386 | 29.9560 | 21.9138 | 0.4376 | 0.2620 | 3.2630 |
Example 3:
As described in Example 1, select the smoking volunteer urine sample being numbered C, record total metals level in twenty-four-hour urine liquid sample as follows:
| Chromium | Nickel | Arsenic | Selenium | Cadmium | Mercury | Plumbous |
| μg | μg | μg | μg | μg | μg | μg |
| 38.0987 | 2.4883 | 12.2719 | 15.0335 | 0.7927 | 0.7445 | 2.3468 |
Claims (1)
1. the assay method of chromium, nickel, arsenic, selenium, cadmium, mercury, lead content in urine, is characterized in that: comprise following processing step:
A. urine capture, gathers smoking, and whole urine capture that non-smoking volunteer drains for 24 hours is also preserved;
B. sample preparation, by urine normal temperature unfreezing, and on the oscillator by urine concussion evenly, getting 10mL urine pours in micro-wave diminishing pot, add 65% red fuming nitric acid (RFNA) and hydrogen peroxide, then put into microwave dissolver and carry out micro-wave digestion, micro-wave digestion heating schedule is: intensification 20min to 95 DEG C, keeps 8min; Intensification 15min to 125 DEG C, keeps 8min; Intensification 20min to 165 DEG C, keeps 20min; Lower the temperature 20min to 20 DEG C, keep 20min; Be warming up to 190 DEG C, keep 10min; Sample volume fraction 5%HNO after digestion process completes
3solution is settled to 50mL;
C. the preparation of seven heavy metal species standard solution:
Chromium, nickel, arsenic, selenium, cadmium, the preparation of the serial mixed standard solution of plumbous six heavy metal species: the hybrid standard product 0.500g accurately being pipetted 10 μ g/mL by electronic balance, 50.000g is settled to the salpeter solution of volume fraction 5%, concentration 100.000 μ g/L after constant volume, pipette the mixed standard solution 0 of 100.000 μ g/L respectively, 0.250, 0.500, 1.000, 2.000, 5.000, 10.000g, dilute with the dust technology of V/V5%, balance is used to be settled to 50.000, 50.000, 50.000, 50.000, 40.000, 50.000, 50.000g, shake up, join respectively 0, 0.500, 1.000, 2.000, 5.000, 10.000, the serial mixed standard solution of 20.000 μ g/L,
The preparation of mercury standard solution: the mercury standard items 0.500g accurately being pipetted 10 μ g/mL by electronic balance, 50.000g is settled to the salpeter solution of volume fraction 5%, concentration 100.000 μ g/L after constant volume, pipette respectively the mercury standard solution 0 of 100.000 μ g/L, 0.250,0.500,1.000,2.000g, dilute with the dust technology of V/V5%, use balance to be settled to 50.000,50.000,50.000,50.000,40.000g, join to obtain the serial mercury standard solution of 0,0.500,1.000,2.000,5.000 μ g/L respectively;
Sample determination, inductivity coupled plasma mass spectrometry measures, draw the heavy metal standard solution of the variable concentrations prepared, using argon gas as carrier, at high temperature the charged steam of standard solution with heavy metal element is blown into inductivity coupled plasma mass spectrometry to measure, be that 52 places record chromium in mass-to-charge ratio, 60 places record nickel, 75 places record arsenic, 82 places record selenium, 111 places record cadmium, 202 places record mercury, 208 places record lead, and use scandium, germanium, bismuth, indium is that interior mark corrects instrument, get the sample solution after micro-wave digestion to measure, the signal respective strengths and the concentration known standard solution that record seven Heavy Metallic Elements compare, the content of seven Heavy Metallic Elements in sample can be tried to achieve, in sample determination process, carrier gas argon flow amount is 1.16L/min, and carrier fluid nitric acid concentration of volume percent is 5%.
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