CN103063732A - Method for measuring contents of chromium, nickel, arsenic, selenium, cadmium, mercury and lead in urine - Google Patents
Method for measuring contents of chromium, nickel, arsenic, selenium, cadmium, mercury and lead in urine Download PDFInfo
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Abstract
The invention discloses a method for measuring the contents of chromium, nickel, arsenic, selenium, cadmium, mercury and lead in urine. The method comprises the steps as follows: collecting all urine of a smoker in 24 hours; storing in a polypropylene bucket at the temperature of subzero 80 DEG C; disinfecting the urine with microwaves; taking argon gas as carrier gas under acidic condition; measuring by using inductively coupled plasma mass spectrometry; and comparing with standard series and quantifying. The method has the advantages of accurate measurement result, good sensitivity and repeatability and the like, and is simple, convenient and quick to operate.
Description
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Technical field:
The invention belongs to the collection analysis detection technique field of urine, relate generally to trace element determination techniques field in the 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 can along with main flume enters human body, cause certain harm to human body as harmful element important in the cigarette smoke.In the process of smoking property of cigarette, the heavy metal element in the pipe tobacco can be used as flue gas aerosol and is scattered in main flume and the side-stream smoke when cigarette burning, in vivo accumulation after it enters in the body.By the Fenton reaction, generate free radical, free radical can be combined with DNA, causes the oxidative damage of DNA.Heavy metal is considered to cause the major reason of a lot of diseases.
Wang Xiaoyan etc. utilize inductivity coupled plasma mass spectrometry (ICP-MS), have set up the method for the 7 Heavy Metallic Elements content such as Cr, Mn, As, Se, Cd, Hg, Pb in direct dilution metering blood of human body and the urine.Abundant had once waited with aas determination lead, cadmium, copper level in crowd's blood of being exposed to the electronic waste processing environment and the urine, and analyze and research subject interior heavy metal level and Correlative Influence Factors thereof.
At present research smoking there is not yet report to smoker's heavy metal of body level affects.Utilize atomic absorption spectrum can not detect simultaneously fast and accurately trace heavy metal element in multiple smoker's urine.Impact of heavy metals in the smoker's body that causes for smoking needs to get rid of the heavy metal element that exists in diet and the environment to the impact of smoker's heavy metal of body.Therefore the interference of getting rid of diet and environment in the urine capture process is very crucial.Summarize described, assay method when being necessary to set up in a kind of urine detection device for multi metallic elements content.Be used for getting rid of diet and environment for the interference of smoker's heavy metal of body element, and detect simultaneously fast and accurately detection device for multi metallic elements.
Summary of the invention:
Purpose of the present invention is intended to set up seven heavy metal species (chromium in a kind of employing inductively coupled plasma mass spectrometric determination smoker urine, nickel, arsenic, selenium, cadmium, mercury and lead) method of content, the method can be carried out Accurate Determining to seven heavy metal species content increases in smoker's body simultaneously, and measurement result accurately, measure disturb few.
The objective of the invention is to be achieved through the following technical solutions: the assay method of seven heavy metal species content comprises following processing step in the mensuration smoker urine of the present invention:
A, urine capture are divided into two groups of (smokings with the volunteer; Non-smoking), gather whole urines of volunteer in continuous 24 hours.
B, sample preparation, urine is measured 10 mL in the polypropylene bucket, inserts in the micro-wave diminishing pot, then puts into microwave dissolver and carries out micro-wave digestion.
The micro-wave digestion heating schedule that adopts is: 20min to 95 ℃ of intensification keeps 8min; 15min to 125 ℃ of intensification keeps 8min; 20min to 165 ℃ of intensification keeps 20min; Lower the temperature 20min to 20 ℃, keep 20min; Be warming up to 190 ℃, 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 that accurately pipettes 10 μ g/mL by electronic balance, use the 5%(volume fraction) salpeter solution, use balance accurately to be settled to 50.000g, concentration 100.000 μ g/L behind the constant volume.Pipette respectively mixed standard solution 0,0.250,0.500,1.000,2.000,5.000,10.000 g of 100.000 μ g/L with balance, use 5%(V/V) the dilution of rare nitric acid, use balance to be settled to 50.000,50.000,50.000,50.000,40.000,50.000,50.000 g, shake up, join respectively to get the serial mixed standard solution of 0,0.500,1.000,2.000,5.000,10.000,20.000 μ g/L.
Preparation contains the series standard solution of mercury: accurately pipette the mercury standard items 0.500g of 10 μ g/mL by electronic balance, use the 5%(volume fraction) salpeter solution be settled to 50.000g, concentration 100.000 μ g/L behind the use balance constant volume.Pipette respectively mercury standard solution 0,0.250,0.500,1.000,2.000 g of 100.000 μ g/L, use 5%(V/V) the dilution of rare nitric acid, use balance to be settled to 50.000,50.000,50.000,50.000,40.000 g.Join respectively to get the mercury standard solution of 0,0.500,1.000,2.000,5.000 μ g/L.
D, sample determination, inductivity coupled plasma mass spectrometry is measured, the heavy metal standard solution of the variable concentrations that absorption prepares, with argon gas as carrier, at high temperature will be blown into inductivity coupled plasma mass spectrometry with the charged steam of the standard solution of heavy metal element measures, 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, 111 places record cadmium, 202 places record mercury, and 208 places record lead, and use scandium, germanium, bismuth, indium are that interior mark is proofreaied and correct instrument, and the sample solution of getting behind the micro-wave digestion is measured, the signal respective strengths and the concentration known standard solution that record seven Heavy Metallic Elements compare, and can try to achieve the content of seven Heavy Metallic Elements in the sample.
Carrier gas in the 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
The volunteer is divided into two groups of (smokings; Non-smoking) gather whole urines of volunteer in continuous 24 hours.
2. micro-wave digestion sample
Accurately measure urine 10 mL in the polypropylene bucket, place counteracting tank.Use first 1 mL, 30 %(v/v) hydrogen peroxide, 2.5 mL 65%(v/v) HNO
3Add counteracting tank, the program of clearing up that employing table 1 sets is carried out digestion process.After (liquid that counteracting tank is packed at most) clears up end, after temperature is down to room temperature, take out counteracting tank, then digestion solution be transferred in the 50mL volumetric flask, use 5%(v/v) HNO
3Clean counteracting tank, and use 5%(v/v) HNO
3Be settled to 50mL, to be measured.Do blank test in company with sample.
Table 1 micro-wave digestion program
| Stage | 1 | 2 | 3 | 4 | 5 |
| Operation energy (W) | 1200 | 1200 | 1200 | 1200 | 1200 |
| Heating-up time (min) | 20 | 15 | 20 | 20 | 20 |
| Terminal temperature (℃) | 95 | 125 | 165 | 20 | 190 |
| Retention time (min) | 8 | 8 | 15 | 20 | 10 |
3. inductivity coupled plasma mass spectrometry is measured
3.1 inductivity coupled plasma mass spectrometry parameter
The experiment condition of inductivity coupled plasma mass spectrometry is as shown in table 2:
Table 2 cold atom absorption spectrum instrument parameter
| Parameter | Condition |
| Measure element | Chromium, nickel, arsenic, selenium, cadmium, mercury, lead |
| Radio frequency intensity | 1330w |
| The sample degree of depth | 7.5mm |
| Flow rate of carrier gas | 1.16L/min |
| Pump speed | 0.10rps |
3.2 the preparation of seven heavy metal species standard solution:
The preparation of six heavy metal species hybrid standard working solutions: the hybrid standard product 0.500g that accurately pipettes 10 μ g/mL by electronic balance, use the 5%(volume fraction) salpeter solution, use balance accurately to be settled to 50.000g, concentration 100.000 μ g/L behind the constant volume.Pipette respectively mixed standard solution 0,0.250,0.500,1.000,2.000,5.000,10.000 g of 100.000 μ g/L with balance, use 5%(V/V) the dilution of rare nitric acid, use balance to be settled to 50.000,50.000,50.000,50.000,40.000,50.000,50.000 g, shake up, join respectively to get the serial mixed standard solution of 0,0.500,1.000,2.000,5.000,10.000,20.000 μ g/L.
The preparation of mercury standard operation solution: accurately pipette the mercury standard items 0.500g of 10 μ g/mL by electronic balance, use the 5%(volume fraction) salpeter solution is settled to 50.000g, uses concentration 100.000 μ g/L behind the balance constant volume.Pipette respectively amalgamation standard solution 0,0.250,0.500,1.000,2.000 g of 100.000 μ g/L, use 5%(V/V) the dilution of rare nitric acid, use balance to be settled to 50.000,50.000,50.000,50.000,40.000 g.Join respectively to get the mercury standard solution of 0,0.500,1.000,2.000,5.000 μ g/L.
3.3 the mensuration of seven Heavy Metallic Elements content
Take argon gas as carrier gas, according to inductivity coupled plasma mass spectrometry: the condition of work of 7500A type is measured the series standard working solution, obtains calibration curve; Mean value with 2 replicate determinations represents test result.Calculate regression equation and the related coefficient of typical curve.The sample that cancellation is separated after processing is measured, and records the signal respective strengths of seven Heavy Metallic Elements, and the substitution regression equation can be obtained the content of seven Heavy Metallic Elements in the sample.
3.4 the repeatability of the inventive method, detection limit and recovery of standard addition
The standard solution of getting 1.000 μ g/L carries out 2 replicate determinations, and relative standard deviation (RSD) is chromium: 4.12%, and nickel: 1.57%, arsenic: 2.61%, selenium: 1.00%, cadmium: 3.17%, mercury: 0.44%, lead, 1.81%.Simultaneously, according to this method blank solution is measured, it is as shown in table 3 to obtain its detection limit with 3 times of standard deviations.
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, by the condition after optimizing the content of heavy metal in the urine has been carried out measuring and having investigated the recovery of method, as shown in table 4.
Table4
Seven heavy metal species assay and recovery of standard addition in the urine
The present invention has set up the mensuration of seven Heavy Metallic Elements and the method for accurate preparing standard solution in the urine, proposes the effective collect means to smoker's urine.And use the inductively coupled plasma mass spectroscopy to detect simultaneously fast and accurately seven Heavy Metallic Elements.The inventive method has following excellent results:
1. the volunteer is divided into two groups of (smokings; Non-smoking), gather twenty-four-hour urine liquid.
2. adopt the micro-wave digestion mode can reduce sample and strong acid use amount, improve and to clear up ability, and sample element loss to be measured few, operating personnel are endangered little, easy operating.Digestion solution clarification, transparent, precipitation not.
3. adopt the quantitative preparing standard solution of electronic balance, than using volumetric flask preparing standard solution, 3 orders of magnitude of accuracy Effective Raise.
When 4. adopting the inductively coupled plasma mass spectrum that the mercury in the cigarette mainstream flue gas is measured, it is high to detect the recovery, and measurement result is accurate.
5. this method has the advantage of easy and simple to handle, quick, accurate, sensitivity and good reproducibility.
Description of drawings
Fig. 1 is smoker's urine sample collecting flowchart figure of the present invention (this figure is as Figure of abstract).
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Embodiment
The present invention is described further below in conjunction with example, but is not restriction the present invention.
The volunteer is divided into two groups of (smokings; Non-smoking), gather twenty-four-hour urine liquid.
After collection was finished, sample preparation was accurately measured urine 10 mL, inserts in the micro-wave diminishing pot, then puts into microwave dissolver and carries out micro-wave digestion.(liquid that counteracting tank is packed at most)
After clearing up end, after temperature is down to room temperature, take out counteracting tank, clear up end after, after temperature is down to room temperature, take out counteracting tank, then digestion solution is transferred in the 50mL volumetric flask, use 5%(v/v) HNO
3Clean counteracting tank, and use 5%(v/v) HNO
3Be settled to 50mL, to be measured.Do blank test in company with sample.
Sample determination, inductivity coupled plasma mass spectrometry is measured, the heavy metal standard operation solution of the variable concentrations that absorption prepares, at high temperature will be blown into inductivity coupled plasma mass spectrometry with the charged steam of heavy metal atom sample and measure as carrier with argon gas.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 the use scandium, germanium, bismuth, indium are interior scalar quantity.Sample solution after cancellation is separated is measured, and the intensity and the standard items that record seven Heavy Metallic Elements compare, and can try to achieve the content of seven Heavy Metallic Elements in the sample.
Example 1:
1. instrument and reagent: inductively coupled plasma: Agilent company, 7500A type inductivity coupled plasma mass spectrometry; Mars 5 type microwave dissolvers (U.S. CE M company) are joined the high-pressure sealed counteracting tank of PTFE; Electronic balance, CP225D type, sensibility reciprocal 0.0001g(Sartorius AG company); Argon gas: purity is 99.99%;
Ultrapure water.Mass percent is the red fuming nitric acid (RFNA) of 65 %, and mass percent is that 30% hydrogen peroxide is chromatographically pure; Hybrid standard product and mercury standard items (10.0 mg/L, U.S. Agilent company).
2. sample preparation: urine is measured 10 mL in the polypropylene bucket, inserts in the micro-wave diminishing pot, then puts into microwave dissolver and carries out micro-wave digestion.(liquid that counteracting tank is packed at most) uses 1 mL, 30 %(v/v successively) hydrogen peroxide, the red fuming nitric acid (RFNA) of 2.5 mL, 65 % is inserted in the counteracting tank, then puts into microwave dissolver, and the program of clearing up that employing table 1 sets is carried out digestion process.After (because counteracting tank pack at most liquid) clears up end, after temperature is down to room temperature, take out counteracting tank, then digestion solution is transferred in the 50mL volumetric flask and constant volume to be measured.
3. assay method:
With carrier gas (argon gas) flow 1.16 L/min, open argon gas, recirculated water, inductivity coupled plasma mass spectrometry, open working software, by table 2 setting the mass spectrum parameter is set, absorption standard blank solution, 0,0.500,1.000,2.000,5.000,10.000, the 20.000 μ g/L hybrid standard working solutions that prepare and clear up after sample enter mass spectrometer and measure.It is as follows to record in the twenty-four-hour urine liquid sample seven heavy metal species total amounts:
| 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 to be numbered the smoking volunteer urine sample of B, it is as follows to record twenty-four-hour urine liquid sample total metals:
| 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 to be numbered the smoking volunteer urine sample of C, it is as follows to record in the twenty-four-hour urine liquid sample total metals level:
| 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 (4)
1. the assay method of chromium, nickel, arsenic, selenium, cadmium, mercury, lead content in the urine is characterized in that: comprise following processing step:
A, urine capture gather whole urine captures and preservation that volunteer's (smoking, non-smoking) drained in 24 hours;
B, sample preparation, with the urine normal temperature unfreezing, and on oscillator that the urine concussion is even, get 10 mL urines and pour in the 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; Sample 5%(massfraction after digestion process is finished) HNO
3Solution is settled to 20 mL;
C,The preparation of seven heavy metal species standard solution:
Preparation contains the serial mixed standard solution of chromium, nickel, arsenic, selenium, cadmium, plumbous six heavy metal species: concentration is respectively: 0,0.500,1.000,2.000,5.000,10.000,20.000 μ g/L;
Preparation contains the series standard solution of mercury: concentration is respectively 0,0.500,1.000,2.000,5.000 μ g/L;
D, sample determination, inductivity coupled plasma mass spectrometry is measured, the heavy metal standard solution of the variable concentrations that absorption prepares, with argon gas as carrier, at high temperature will be blown into inductivity coupled plasma mass spectrometry with the charged steam of the standard solution of heavy metal element measures, 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, 111 places record cadmium, 202 places record mercury, and 208 places record lead, and use scandium, germanium, bismuth, indium are that interior mark is proofreaied and correct instrument, and the sample solution of getting behind the micro-wave digestion is measured, the signal respective strengths and the concentration known standard solution that record seven Heavy Metallic Elements compare, and can try to achieve the content of seven Heavy Metallic Elements in the sample.
2. the assay method of chromium, nickel, arsenic, selenium, cadmium, mercury, lead content in the urine according to claim 1 is characterized in that:
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 that accurately pipettes 10 μ g/mL by electronic balance, use the 5%(volume fraction) salpeter solution be settled to 50.000g, concentration 100.000 μ g/L behind the constant volume, pipette respectively the mixed standard solution 0 of 100.000 μ g/L, 0.250,0.500,1.000,2.000,5.000,10.000 g, use 5%(V/V) the dilution of rare nitric acid, use balance to be settled to 50.000,50.000,50.000,50.000,40.000,50.000,50.000, shake up, join respectively 0,0.500,1.000,2.000,5.000,10.000,20.000 the serial mixed standard solution of μ g/L;
The preparation of mercury standard solution: the mercury standard items 0.500g that accurately pipettes 10 μ g/mL by electronic balance, use the 5%(volume fraction) salpeter solution be settled to 50.000g, concentration 100.000 μ g/L behind the constant volume, pipette respectively mercury standard solution 0,0.250,0.500,1.000,2.000 g of 100.000 μ g/L, use 5%(V/V) the dilution of rare nitric acid, use balance to be settled to 50.000,50.000,50.000,50.000,40.000 g, join respectively to get the serial mercury standard solution of 0,0.500,1.000,2.000,5.000 μ g/L.
3. the assay method of chromium, nickel, arsenic, selenium, cadmium, mercury, lead content in the urine according to claim 1, it is characterized in that: the micro-wave digestion heating schedule is: 20min to 95 ℃ of intensification keeps 8min; 15min to 125 ℃ of intensification keeps 8min; 20min to 165 ℃ of intensification keeps 20min; Lower the temperature 20min to 20 ℃, keep 20min; Be warming up to 190 ℃, keep 10min.
4. the assay method of chromium, nickel, arsenic, selenium, cadmium, mercury, lead content in the urine according to claim 1, it is characterized in that: carrier gas in the sample determination process (argon gas) flow is 1.16L/min, carrier fluid (nitric acid) concentration of volume percent is 5%.
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