Background technology
In recent years, along with the raising of China's industrial level, the kind of poisonous or potential poisonous chemical, particularly heavy metal and quantity increase to some extent in the environment, and humans and animals has been constituted serious threat, and wherein the harm of cadmium and lead is especially outstanding.Cadmium is the secondary product that extracts zinc, is used for plating, alloy manufacturing field more, studies show that, cadmium poisoning can cause renal tubule malabsorption again, cause low molecular weight protein and calcareous etc., go down to form easily osteomalacia, arthralgia, fracture and textured bone etc. for a long time by running off in the urine.Lead is a kind of neurovirulent heavy metal element that has, because all there is a certain amount of lead in the ubiquity of lead in the environment in most human bodies, plumbous amount in vivo surpasses certain level will cause infringement to health.Children are because metabolism and development are responsive especially to Lead Toxicity.Studies confirm that Pb-B though still be not enough to produce specific clinical manifestation, can have a negative impact to children's intellectual development, Physique growth, learning ability and hearing when 10 μ g/dL (the 0.483 μ moL/L) left and right sides.Therefore, reduce plumbous and cadmium harm at present putting prevention first, and accurately detect the lead in the blood, the content of cadmium element is the first step that the plumbous cadmium of prevention endangers.
At present mainly adopt flameless atomic absorption spectrometry for the detection of the lead and cadmium elements in the human whole blood, instrument is traditional graphite furnace atomic absorption spectrophotometer, and graphite furnace atomic absorption spectrophotometer requires high to operating environment, and complex operation, the use cost height.For addressing the above problem, my company's independent development tungsten boat Atomic Absorption Spectrometer (obtained Chinese patent, the patent No. 02209376.1), primary structure comprises element lamp, lens behind the element lamp, the multiple tracks monochromator in the band light hole behind the lens, the information detector that links to each other with the multiple tracks monochromator, the singlechip controller that links to each other with information detector is promptly by the connected microcomputer of Serial Port Line, also be provided with tungsten boat atomizer between lens and the multiple tracks monochromator, tungsten boat atomizer is by the perforation of band symmetry, the round metal column casing of inert gas import, the thermal conductor tungsten sheet that sets up between pair of electrodes that is provided with in the tube and the electrode is formed, and thermal conductor tungsten sheet center section forms groove and is the boat type.This patent can single injected sampling, finish the lead in the sample simultaneously, the measurement of two kinds of elements of cadmium, shortened detection time, improved measuring accuracy, but disturb for preventing to produce in the whole blood, and guarantee all to be in the calibration curve range of linearity after the concentration of element dilution to be measured, need carry out matrix modification to whole blood, traditional atom analysis method amount for taking blood is bigger, generally more than 2mL, colonies such as children are easily caused big painful, though can adopt the method for microwave sample dissolving to do sample pre-treatments, reduce amount for taking blood, the instrument costliness, complex operation step and have certain danger is to testing agency's environment such as hospitals, peopleware is had relatively high expectations.Therefore need a kind of amount for taking blood of exploitation matrix modifying reagent and using method little, simple to operation, satisfy and reduce the patient suffering, reduce the market demand that hospital conditions is required.
Summary of the invention
The purpose of this invention is to provide a kind of matrix modifying reagent and detect the method for content of lead and cadmium elements in the whole blood with it.
A kind of matrix modifying reagent that is used for detecting the whole blood content of lead and cadmium elements, comprise following component: mass concentration is the diammonium hydrogen phosphate of 5-6 ‰, volumetric concentration is the Triton X-100 of 2.0-4.0 ‰, and volumetric concentration is the red fuming nitric acid (RFNA) of 1.0-3.0 ‰, and all the other are ultrapure water.
Matrix modifying reagent of the present invention can also comprise that volumetric concentration is the calf serum of 0.5-1.5 ‰, detects better effects if.
Matrix modifying reagent of the present invention, preferably form by following component:
Mass concentration is 5.5 ‰ diammonium hydrogen phosphate, and volumetric concentration is 3.0 ‰ Triton X-100, and volumetric concentration is 2.0 ‰ red fuming nitric acid (RFNA), and volumetric concentration is 1.0 ‰ calf serum, and all the other are ultrapure water.
With above-mentioned preferred matrix modifying reagent is example, its preparation method can for: take by weighing 11.00 gram top grade pure phosphoric acid hydrogen, two ammoniums, place the 2000mL volumetric flask, add 6.0mL Triton X-100,4.0mL red fuming nitric acid (RFNA) and 2.0mL calf serum, be settled to scale with ultrapure water again.
The present invention also provides the method that detects content of lead and cadmium elements in the whole blood with described matrix modifying reagent, adopts described matrix modifying reagent that no NITRATE BY FLAME ATOMIC absorption plant detection is gone up in 1/8th to 1/13rd backs that whole blood is diluted to original content.Wherein atomic absorption unit preferably adopts tungsten boat Atomic Absorption Spectrometer.Especially preferably the BH2100S/T type Atomic Absorption Spectrometer that adopts the applicant (Bohui Innovation Photoelectric Technology Co. Ltd., Beijing) to produce, it has a detailed description in Chinese patent 02209376.1.
Technique scheme has following advantage:
1, amount for taking blood is few.Compare with classic method, only need get 10-50 microlitre whole blood and carry out the matrix improvement, get final product content plumbous in the while measuring samples, cadmium two elements, reduced the amount for taking blood of testing agency, reduce the patient suffering, help business promotion.
2, adopt preferred ratio preparation back to pack in the centrifuge tube, preserves standbyly, directly in centrifuge tube, add whole blood during use, do not need in the high standard laboratory, to carry out the solution preparation, environment and operating personnel's requirement reduction by the standard specification branch.
3, adopt unified raw material, operation preparation, can, error is little between each dilution, the systematic error that has reduced to exist in hemodilution, the processes such as solution preparation before need not detecting, convenient, the safety of method of operating.
Embodiment
Below in conjunction with embodiment, the specific embodiment of the present invention is described in further detail.Following examples are used to illustrate the present invention, but are not used for limiting the scope of the invention.
Embodiment 1
Matrix modifying reagent and preparation thereof and using method:
The matrix modifying reagent prescription is as follows: diammonium hydrogen phosphate 5 ‰, Triton X-100 2.0 ‰, red fuming nitric acid (RFNA) 1.0 ‰, all the other are ultrapure water.
Preparation: take by weighing 10.00 gram top grade pure phosphoric acid hydrogen, two ammoniums, place the 2000mL volumetric flask, add 4.0mL Triton X-100,2.0mL red fuming nitric acid (RFNA), be settled to scale with ultrapure water again, be filled to centrifuge tube by the specification branch and get product.
With its method that detects content of lead and cadmium elements in the whole blood, may further comprise the steps:
(1) get the centrifuge tube of the matrix modifying reagent that 0.07mL is housed, standby;
(2) under 10 ℃-30 ℃, in the centrifuge tube that matrix modifying reagent is housed, add the whole blood of 10 μ L, cover tight centrifuge tube lid, the mixing that vibrates rapidly makes whole blood be diluted to 1/8th of original content, and is static to be checked.
(3) go up the BH2100S/T type Atomic Absorption Spectrometer detection that the Bohui Innovation Photoelectric Technology Co. Ltd., Beijing produces.
Embodiment 2
Matrix modifying reagent and preparation thereof and using method:
The matrix modifying reagent prescription is as follows: diammonium hydrogen phosphate 5.5 ‰, Triton X-100 3.0 ‰, red fuming nitric acid (RFNA) 2.0 ‰, all the other are ultrapure water.
The preparation method is with embodiment 1.
With its method that detects content of lead and cadmium elements in the whole blood, may further comprise the steps:
(1) get the centrifuge tube of the matrix modifying reagent that 0.09mL is housed, standby;
(2) under 10 ℃-30 ℃, in the centrifuge tube that matrix modifying reagent is housed, add the whole blood of 10 μ L, cover tight centrifuge tube lid, the mixing that vibrates rapidly makes whole blood be diluted to 1/10th of original content, and is static to be checked;
(3) go up BH2100S/T type Atomic Absorption Spectrometer (wherein S represents to mould shell, and T the represents iron-clad) detection that the Bohui Innovation Photoelectric Technology Co. Ltd., Beijing produces.
Embodiment 3
Matrix modifying reagent and preparation thereof and using method:
The matrix modifying reagent prescription is as follows: diammonium hydrogen phosphate 6 ‰, Triton X-100 4.0 ‰, red fuming nitric acid (RFNA) 3.0 ‰, all the other are ultrapure water.
The preparation method is with embodiment 1.
With its method that detects content of lead and cadmium elements in the whole blood, may further comprise the steps:
(1) get the centrifuge tube of the matrix modifying reagent that 0.12mL is housed, standby;
(2) under 10 ℃-30 ℃, in the centrifuge tube that matrix modifying reagent is housed, add the whole blood of 10 μ L, cover tight centrifuge tube lid, the mixing that vibrates rapidly makes whole blood be diluted to 1/13rd of original content, and is static to be checked;
(3) go up the BH2100S/T type Atomic Absorption Spectrometer detection that the Bohui Innovation Photoelectric Technology Co. Ltd., Beijing produces.
Embodiment 4
Matrix modifying reagent and preparation thereof and using method:
The matrix modifying reagent prescription is as follows: diammonium hydrogen phosphate 5 ‰, Triton X-100 2.0 ‰, red fuming nitric acid (RFNA) 1.0 ‰, calf serum 0.5 ‰, all the other are ultrapure water.
Preparation: take by weighing 10.00 gram top grade pure phosphoric acid hydrogen, two ammoniums, place the 2000mL volumetric flask, add 4.0mL Triton X-100,2.0mL red fuming nitric acid (RFNA), 1.0mL calf serum, be settled to scale with ultrapure water again, be filled to centrifuge tube by the specification branch and get product.
With its method that detects content of lead and cadmium elements in the whole blood, may further comprise the steps:
(1) get the centrifuge tube of the matrix modifying reagent that 0.28mL is housed, standby;
(2) under 10 ℃-30 ℃, in the centrifuge tube that matrix modifying reagent is housed, add the whole blood of 40 μ L, cover tight centrifuge tube lid, the mixing that vibrates rapidly makes whole blood be diluted to 1/8th of original content, and is static to be checked.
(3) go up the BH2100S/T type Atomic Absorption Spectrometer detection that the Bohui Innovation Photoelectric Technology Co. Ltd., Beijing produces.
Embodiment 5
Matrix modifying reagent and preparation thereof and using method:
The matrix modifying reagent prescription is as follows: diammonium hydrogen phosphate 5.5 ‰, Triton X-100 3.0 ‰, red fuming nitric acid (RFNA) 2.0 ‰, calf serum 1.0 ‰, all the other are ultrapure water.
The preparation method is with embodiment 4.
With its method that detects content of lead and cadmium elements in the whole blood, may further comprise the steps:
(1) get the centrifuge tube of the matrix modifying reagent that 0.36mL is housed, standby;
(2) under 10 ℃-30 ℃, in the centrifuge tube that matrix modifying reagent is housed, add the whole blood of 40 μ L, cover tight centrifuge tube lid, the mixing that vibrates rapidly makes whole blood be diluted to 1/10th of original content, and is static to be checked;
(3) go up the BH2100S/T type Atomic Absorption Spectrometer detection that the Bohui Innovation Photoelectric Technology Co. Ltd., Beijing produces.
Embodiment 6
Matrix modifying reagent and preparation thereof and using method:
The matrix modifying reagent prescription is as follows: diammonium hydrogen phosphate 6 ‰, Triton X-100 4.0 ‰, red fuming nitric acid (RFNA) 3.0 ‰, calf serum 1.5 ‰, all the other are ultrapure water.
The preparation method is with embodiment 4.
With its method that detects content of lead and cadmium elements in the whole blood, may further comprise the steps:
(1) get the centrifuge tube of the matrix modifying reagent that 0.48mL is housed, standby;
(2) under 10 ℃-30 ℃, in the centrifuge tube that matrix modifying reagent is housed, add the whole blood of 40 μ L, cover tight centrifuge tube lid, the mixing that vibrates rapidly makes whole blood be diluted to 1/13rd of original content, and is static to be checked;
(3) go up the BH2100S/T type Atomic Absorption Spectrometer detection that the Bohui Innovation Photoelectric Technology Co. Ltd., Beijing produces.
Test example 1:
The matrix modifying reagent that adopts embodiment 2 detects content of lead and cadmium elements to going up the BH2100S/T type Atomic Absorption Spectrometer that the Bohui Innovation Photoelectric Technology Co. Ltd., Beijing produces behind the whole blood dilution different proportion respectively, and testing result sees Table 1 and table 2 respectively:
The matrix modifying reagent of table 1: embodiment 2 is to plumbous testing result (μ g/L)
The matrix modifying reagent of table 2: embodiment 2 is to cadmium testing result (μ g/L)
Test example 2:
Adopt the matrix modifying reagent of embodiment 5 to detect content of lead and cadmium elements to going up tungsten boat Atomic Absorption Spectrometer behind the whole blood dilution different proportion respectively, testing result sees Table 3 and table 4 respectively:
The matrix modifying reagent of table 3: embodiment 5 is to plumbous testing result (μ g/L)
The matrix modifying reagent of table 4: embodiment 5 is to cadmium testing result (μ g/L)
From table 1, table 2, table 3 and table 4 as can be seen, matrix modifying reagent provided by the invention can be measured the content of lead and cadmium elements in the whole blood exactly, if wherein pack contains calf serum, measurement effect is better; Adopt matrix modifying reagent provided by the invention, whole blood is diluted to 1/8th to 1/13rd of original content, can make the limits of calibration curve when the content of lead and cadmium elements meets mensuration in the blood, can disposablely measure content plumbous and cadmium element simultaneously, and the measuring accuracy height is the best to be diluted to 1/10th of original content especially wherein.
Atomic absorption spectrophotometry is a kind of common blood testing, and it is strong to have selectivity, disturb less, accuracy height, the advantage that analysis speed is fast.In the real work, in order to guarantee the whole blood stable in properties, satisfy atomic absorption spectrum and detect requirement, needed blood is carried out matrix modification before blood measuring, the prior art amount for taking blood is big, and complex operation requires height to environment, operating personnel.The present invention adopts the matrix modifying reagent of optimized proportion, and get the centrifuge tube that 7-12 part whole blood matrix modifying reagent is housed, under 10-30 ℃, the whole blood that in the centrifuge tube that matrix modifying reagent is housed, adds 1 volume parts, cover tight centrifuge tube lid, the vibration mixing makes whole blood be diluted to 1/8th to 1/13rd of original content rapidly, and is static to be checked.The present invention has the instrument of meeting and human detection requirement because of adopting optimized proportion by the standardized size packing, and practical operation is convenient, the characteristics that systematic error is little.
The above only is a preferred implementation of the present invention; should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the technology of the present invention principle; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.