CN108414502A - A kind of method of micronutrient levels in measurement blood vessel - Google Patents
A kind of method of micronutrient levels in measurement blood vessel Download PDFInfo
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- 239000011785 micronutrient Substances 0.000 title claims abstract description 29
- 210000004204 blood vessel Anatomy 0.000 title claims abstract description 19
- 235000013369 micronutrients Nutrition 0.000 title claims abstract description 19
- 238000000034 method Methods 0.000 title claims abstract description 12
- 238000005259 measurement Methods 0.000 title description 5
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims abstract description 65
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 claims abstract description 40
- 239000011777 magnesium Substances 0.000 claims abstract description 40
- 229910052749 magnesium Inorganic materials 0.000 claims abstract description 40
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims abstract description 39
- 239000011575 calcium Substances 0.000 claims abstract description 39
- 229910052791 calcium Inorganic materials 0.000 claims abstract description 39
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- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 claims abstract description 32
- 229910052742 iron Inorganic materials 0.000 claims abstract description 32
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/71—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light thermally excited
- G01N21/73—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light thermally excited using plasma burners or torches
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- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
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- Immunology (AREA)
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- Investigating Or Analysing Biological Materials (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention discloses a kind of analysis method measuring micro- (phosphorus, calcium, magnesium, iron, zinc) content in blood vessel using inductive coupling plasma emission spectrograph.Method uses nitric acid hydrogen peroxide Acid system, and through wet digestion, standard curve is established, then measures five kinds of phosphorus, calcium, magnesium, iron, zinc micronutrient levels simultaneously by inductive coupling plasma emission spectrograph.The present invention is to probe into the content of trace element to the elasticity of blood vessel, the damage of vascular endothelial cell, reparation and Apoptosis, and then find out interaction rule between trace element and vascular diseases and provide important data analysis detection foundation.
Description
Technical field
The present invention relates to a kind of points measuring micro- (phosphorus, calcium, magnesium, iron, zinc) content in blood vessel using ICP-OES methods
Analysis method, belongs to analysis technical field.
Background technology
Blood vessel is hemophoric pipeline, can be divided into artery, vein and capilary according to transporting direction.Aorta and main artery
Tube wall it is thicker, containing abundant elastomer, there is expansivity and elasticity；The bore of capillary is minimum, and quantity is most,
Total cross-sectional area is maximum, and blood flow velocity is most slow, and tube wall is most thin, is only made of monolayer endothelial cell and basement membrane, and permeability is fine,
Be conducive to blood and carry out mass exchange with tissue；Vein is compared with corresponding artery, and quantity is big, and bore is big, and tube wall is thin, is easily expanded
, it is called capacity vessel.Common vascular diseases refer mainly to atherosclerosis, inflammatory vascular diseases, functional vascular disease
Disease, the true neoplasm disease etc. of blood vessel.
Calcium in addition to be skeleton development base stock, also in vivo have other important physiological functions.Calcium can adjust enzyme
Activity, participate in nerve, the activity of muscle and the release of neurotransmitter, the secretion for adjusting hormone, adjust the rhythm of the heart, reduce it is cardiovascular
Permeability, control inflammation with oedema, maintain acid-base balance etc..Calcium deficiency can lead to aging, osteoporosis, allergy, oedema, blood vessel
Hardening etc..
Phosphorus is the important composition material of bone tooth, facilitates bone and the indispensable nutrient of tooth calcification, is composition
One of the basis of inhereditary material nucleic acid, participates in the adjusting of internal acid-base balance, participates in the metabolism of energy i (in vivo).Phosphorus is taken in
Or hypophosphatemia can occur in the deficiency absorbed, cause the exception of red blood cell, leucocyte, blood platelet, osteomalacia, hyperphosphatemia,
Blood calcium reduction in blood is set to lead to osteoporosis.
Iron is the indispensable trace element of human body.It has the function of fixed oxygen and conveying oxygen, improves immunity of organisms.
Human body iron deficiency can cause anemia, the damage and behavior change of psychological activity and intellectual development.Fe supply will produce heart disease,
It is related with the tumour of a variety of organs such as liver, colon, rectum, lung, oesophagus, bladder.
Magnesium is the dominant cation in human body cell, and participating in organism normal activities and metabolic processes must can not
Few element.Magnesium influences a variety of biological functions of cell, also participates in the stability for maintaining genome, and also answered with body oxidation
Swash related with tumour generation.Magnesium shortage can cause serum calcium to decline, and neural muscle excitability is hyperfunction, may have to vascular function potential
Influence, magnesium plays an important role to the homeostasis of bone mineral, magnesium lack may be Postmenopausal Osteoporosis a kind of danger
Factor.
Zinc is one of the essential trace elements of the human body, important in growth in humans's development, reproduction heredity, immune, endocrine etc.
It plays an important role in physiology course.Zinc deficiency easily causes nanism, feeblemindedness, immune function decline, reproduction
System function is impaired, wound healing is slow, easy to catch cold, miscarriage, premature labor, impotence generandi, graying of hair, alopecia, optic nerve wither
Contracting, myopia, cataract, age-related macular degeneration, the elderly accelerate aging, ischemic disease, toxaemia, hepatic sclerosis etc..
Vascular diseases are generated mainly due to vascular wall because lesion loses elasticity, become weak, and pressure effect is born long-term
It is lower to expand, Angioma-like anomalies are generated, or even cause to rupture and bleeding；Lesion makes luminal stenosis, then occurs by for device
The ischemic of official or limbs so that necrosis；Lesion makes to induce intravascular coagulation after blood vessel internal membrane damage, thrombosis, then generator
Blocking remote tube chamber after official or tissue ischemia or embolus fall off.Research worker is exploring all kinds of of influence vascular lesion always
Factor, micro- elasticity of the content to blood vessel, the damage to vascular endothelial cell, reparation and apoptosis are that have in medium vessels
It influences, therefore, the micronutrient levels in Accurate Determining blood vessel is probed into vascular diseases Crack cause and disease treatment has
Important data supporting effect.
The quantitative detection of phosphorus, calcium, magnesium, iron, zinc microelement can be used ultraviolet specrophotometer, Atomic Absorption Spectrometer,
Inductive coupling plasma emission spectrograph or icp ms detection.But UV spectrophotometer measuring member
Plain method is cumbersome, and the Atomic Absorption Spectrometer range of linearity is relatively narrow and multielement can not be carried out at the same time detection, inductively coupled plasma
Constitution spectrometer cost is higher, and is more suitable for trace element detection.
Invention content
Present invention aims at micro in a kind of easy, accurate, high detection blood vessel based on ICP-OES of repeatability of offer
The analysis method of element (phosphorus, calcium, magnesium, iron, zinc) content provides to explore trace element and the interaction relationship of vascular diseases
Theory and technology supports.The present invention is based on trace element with the relevance and inductively-coupled plasma spectrometer of vascular diseases micro-
Secondary element quantifies the superiority of context of detection, establish accurately measured using inductively-coupled plasma spectrometer phosphorus in blood vessel,
The content of calcium, magnesium, iron, zinc microelement, this method have it is easy, can multielement simultaneous determination, be suitble to trace element detection etc.
Advantage.
To achieve the purpose of the present invention, it adopts the following technical scheme that, the specific steps are：
Step 1, the preparation of test sample
(1) the fresh blood vessel samples of 0.5g~1g (being accurate to 0.001g) are accurately weighed in glass digestion utensil, add 2~
4mL top pure grade concentrated nitric acids, and 90~95 DEG C of 5~6h of Water bath (it is slightly cold if color deepens in digestion process, nitric acid is supplemented in right amount
Continue to clear up), 0.5~1mL hydrogen peroxide is added dropwise, until solution is in faint yellow or white clear shape, catches up with acid molten to remaining 1mL or so
Liquid is cooled to room temperature, and 2% nitric acid is used to be settled to 10mL as test solution.
(2) same procedure processing blank reagent is into line blank test, i.e., in the digestion utensil for not healing pipe sample, according to
(1) identical concentrated nitric acid and hydrogen peroxide use are cleared up under the same conditions in, and final constant volume solution is blank solution.
Step 2, Determination of trace elements
(1) mixed standard solution series is prepared：Phosphorus, calcium, magnesium, iron, Zinc standard solution (1000mgL are taken respectively-1) be placed in
In same container, be configured to mix intermediate storing solution with the dilution of 2% nitric acid, phosphorus in storing solution among the mixing, calcium, magnesium, iron,
Zinc concentration is 100mgL-1；The intermediate storing solution of mixing is taken, the mixed standard solution for preparing various concentration is diluted with 2% nitric acid
The concentration of series, the mixed standard solution series is respectively 0.00mgL-1(phosphorus, calcium, magnesium, iron, zinc concentration are
0.00mg·L-1)、0.05mg·L-1(phosphorus, calcium, magnesium, iron, zinc concentration are 0.05mgL-1)、0.10mg·L-1(phosphorus, calcium,
Magnesium, iron, zinc concentration are 0.10mgL-1)、0.50mg·L-1(phosphorus, calcium, magnesium, iron, zinc concentration are 0.50mgL-1)、
1.0mg·L-1(phosphorus, calcium, magnesium, iron, zinc concentration are 1.0mgL-1)、5.0mg·L-1(phosphorus, calcium, magnesium, iron, zinc concentration are
5.0mg·L-1)、10.0mg·L-1(phosphorus, calcium, magnesium, iron, zinc concentration are 10.0mgL-1)。
(2) ICP-OES determination conditions are as follows：
| Operating condition | Parameter |
| Power | 1.1kW |
| Cooling gas flow | 19L/min |
| Atomization gas pressure | 35PSI |
| Secondary air amount | 0.2L/min |
| Pump speed | 1.4mL/min |
(3) standard curve is established：The mixed standard solution series prepared in (1) is injected separately into inductively coupled plasma
Spectrometer, using response as ordinate, draws each element standard curve (P elements spectral line respectively using concentration of element as abscissa：
213.618nm calcium constituent spectral line：317.933nm magnesium elements spectral line：279.553nm ferro element spectral line：238.204nm zinc is first
Plain spectral line：213.856nm).
(4) sample measures：Blank solution and test solution are introduced into inductively-coupled plasma spectrometer respectively successively,
Detection data is obtained, substitutes into the standard curve that (3) are established, it is known that micronutrient levels C in blank solutionIt is emptyAnd in test solution
Micronutrient levels CFor, micronutrient levels in sample can be obtained according to following formula calculating：
In formula, CForMicronutrient levels (mgL in test solution-1)；CIt is emptyMicronutrient levels in blank solution
(mg·L-1)；mSampleSample size (g).
Description of the drawings
Fig. 1 is P elements standard curve；
Fig. 2 is calcium constituent standard curve；
Fig. 3 is magnesium elements standard curve；
Fig. 4 is ferro element standard curve；
Fig. 5 is Zn-ef ficiency standard curve.
Specific implementation mode
It is further illustrated the present invention below by way of specific embodiment, but not as limitation of the present invention.
Phosphorus used in embodiment (GSBG62009-90), calcium (GSBG62012-90), magnesium (GSBG62005-90), iron
(GSBG62020-90), zinc (GSBG62025-90) standard solution (1000mgL-1), derive from Iron and Steel Research Geueral Inst.
Embodiment 1
The measurement of phosphorus, calcium, magnesium, iron, Zn-ef ficiency content, is as follows in a kind of descending aorta straight section：
(1) sample treatment：The accurately weighed fresh blood vessel samples of sample 0.5g add 2mL top pure grades in glass digestion utensil
Concentrated nitric acid, 95 DEG C of Water bath 5h (slightly cold if color deepens in digestion process, the appropriate nitric acid that supplements continues to clear up), is added dropwise
0.5mL hydrogen peroxide is caught up with acid to remaining 1mL or so solution, is cooled to room temperature until solution is in faint yellow or white clear shape, uses
2% nitric acid is settled to 10mL as test solution.
Blank solution is prepared：In the digestion utensil for not healing pipe sample, according to identical concentrated nitric acid and peroxidating in (1)
Hydrogen dosage is cleared up under the same conditions, and final constant volume solution is blank solution.
(2) mixed standard solution series is prepared：Respectively from phosphorus, calcium, magnesium, iron, Zinc standard solution (1000mgL-1) in it is each
5mL solution is pipetted to the same 50mL volumetric flasks, with 2% nitric acid constant volume, be configured to mix intermediate storing solution, among the mixing
Phosphorus, calcium, magnesium, iron, zinc concentration are 100mgL in storing solution-1.A certain amount of solution is pipetted from the intermediate storing solution of mixing, is used
2% nitric acid is settled to 50mL, be configured to concentration be respectively 0.00,0.05,0.10,0.50,1.0,5.0,10.0mgL-1It is mixed
Standardization solution series.
(3) ICP-OES determination conditions are as follows.
| Operating condition | Parameter |
| Power | 1.1kW |
| Cooling gas flow | 19L/min |
| Atomization gas pressure | 35PSI |
| Secondary air amount | 0.2L/min |
| Pump speed | 1.4mL/min |
(4) standard curve is established：The mixed standard solution series prepared in (2) is injected separately into inductively coupled plasma
Spectrometer, using response as ordinate, draws each element standard curve (P elements spectral line respectively using concentration of element as abscissa：
213.618nm calcium constituent spectral line：317.933nm magnesium elements spectral line：279.553nm ferro element spectral line：238.204nm zinc is first
Plain spectral line：213.856nm).
(5) sample measures：Blank solution and test solution are introduced into inductively-coupled plasma spectrometer respectively successively,
Detection data is obtained, substitutes into the standard curve that (3) are established, it is known that micronutrient levels C in blank solutionIt is emptyAnd in test solution
Micronutrient levels CFor, micronutrient levels in sample can be obtained according to following formula calculating：
In formula, CForMicronutrient levels (mgL in test solution-1)；CIt is emptyMicronutrient levels in blank solution
(mg·L-1)；mSampleSample size (g).
As a result it is：Phosphorus 540mgkg-1, calcium 68.1mgkg-1, magnesium 36.0mgkg-1, iron 9.12mgkg-1, zinc
24.2mg·kg-1。
(6) recovery of standard addition：
Embodiment 2
The measurement of phosphorus, calcium, magnesium, iron, Zn-ef ficiency content, is as follows in a kind of descending aorta curved segment：
(1) sample treatment：The accurately weighed fresh blood vessel samples of sample 0.8g add 3mL top pure grades in glass digestion utensil
Concentrated nitric acid, 93 DEG C of Water bath 5.5h (slightly cold if color deepens in digestion process, the appropriate nitric acid that supplements continues to clear up), is added dropwise
0.75mL hydrogen peroxide is caught up with acid to remaining 1mL or so solution, is cooled to room temperature until solution is in faint yellow or white clear shape, uses
2% nitric acid is settled to 10mL as test solution.
Blank solution is prepared：In the digestion utensil for not healing pipe sample, according to identical concentrated nitric acid and peroxidating in (1)
Hydrogen dosage is cleared up under the same conditions, and final constant volume solution is blank solution.
(2) mixed standard solution series is prepared：Respectively from phosphorus, calcium, magnesium, iron, Zinc standard solution (1000mgL-1) in it is each
5mL solution is pipetted to the same 50mL volumetric flasks, with 2% nitric acid constant volume, be configured to mix intermediate storing solution, among the mixing
Phosphorus, calcium, magnesium, iron, zinc concentration are 100mgL in storing solution-1.A certain amount of solution is pipetted from the intermediate storing solution of mixing, is used
2% nitric acid is settled to 50mL, be configured to concentration be respectively 0.00,0.05,0.10,0.50,1.0,5.0,10.0mgL-1It is mixed
Standardization solution series.
(3) ICP-OES determination conditions are as follows：
| Operating condition | Parameter |
| Power | 1.1kW |
| Cooling gas flow | 19L/min |
| Atomization gas pressure | 35PSI |
| Secondary air amount | 0.2L/min |
| Pump speed | 1.4mL/min |
(4) standard curve is established：The mixed standard solution series prepared in (2) is injected separately into inductively coupled plasma
Spectrometer, using response as ordinate, draws each element standard curve (P elements spectral line respectively using concentration of element as abscissa：
213.618nm calcium constituent spectral line：317.933nm magnesium elements spectral line：279.553nm ferro element spectral line：238.204nm zinc is first
Plain spectral line：213.856nm).
(5) sample measures：Blank solution and test solution are introduced into inductively-coupled plasma spectrometer respectively successively,
Detection data is obtained, substitutes into the standard curve that (3) are established, it is known that micronutrient levels C in blank solutionIt is emptyAnd in test solution
Micronutrient levels CFor, micronutrient levels in sample can be obtained according to following formula calculating：
In formula, CForMicronutrient levels (mgL in test solution-1)；CIt is emptyMicronutrient levels in blank solution
(mg·L-1)；mSampleSample size (g).
As a result it is：Phosphorus 365mgkg-1, calcium 48.6mgkg-1, magnesium 13.4mgkg-1, iron 15.2mgkg-1, zinc
32.1mg·kg-1。
(6) recovery of standard addition：
Embodiment 3
The measurement of phosphorus, calcium, magnesium, iron, Zn-ef ficiency content, is as follows in a kind of aorta ascendens straight section：
(1) sample treatment：The accurately weighed fresh blood vessel samples of sample 1.0g add 4mL top pure grades in glass digestion utensil
Concentrated nitric acid, 90 DEG C of Water bath 6h (slightly cold if color deepens in digestion process, the appropriate nitric acid that supplements continues to clear up), is added dropwise 1mL
Hydrogen peroxide is caught up with acid to remaining 1mL or so solution, is cooled to room temperature, with 2% nitre until solution is in faint yellow or white clear shape
Acid is settled to 10mL as test solution.
Blank solution is prepared：In the digestion utensil for not healing pipe sample, according to identical concentrated nitric acid and peroxidating in (1)
Hydrogen dosage is cleared up under the same conditions, and final constant volume solution is blank solution.
(2) mixed standard solution series is prepared：Respectively from phosphorus, calcium, magnesium, iron, Zinc standard solution (1000mgL-1) in it is each
5mL solution is pipetted to the same 50mL volumetric flasks, with 2% nitric acid constant volume, be configured to mix intermediate storing solution, among the mixing
Phosphorus, calcium, magnesium, iron, zinc concentration are 100mgL in storing solution-1.A certain amount of solution is pipetted from the intermediate storing solution of mixing, is used
2% nitric acid is settled to 50mL, be configured to concentration be respectively 0.00,0.05,0.10,0.50,1.0,5.0,10.0mgL-1It is mixed
Standardization solution series.
(3) ICP-OES determination conditions are as follows：
(4) standard curve is established：The mixed standard solution series prepared in (2) is injected separately into inductively coupled plasma
Spectrometer, using response as ordinate, draws each element standard curve (P elements spectral line respectively using concentration of element as abscissa：
213.618nm calcium constituent spectral line：317.933nm magnesium elements spectral line：279.553nm ferro element spectral line：238.204nm zinc is first
Plain spectral line：213.856nm).
(5) sample measures：Blank solution and test solution are introduced into inductively-coupled plasma spectrometer respectively successively,
Detection data is obtained, substitutes into the standard curve that (3) are established, it is known that micronutrient levels C in blank solutionIt is emptyAnd in test solution
Micronutrient levels CFor, micronutrient levels in sample can be obtained according to following formula calculating：
In formula, CForMicronutrient levels (mgL in test solution-1)；CIt is emptyMicronutrient levels in blank solution
(mg·L-1)；mSampleSample size (g).
As a result it is：Phosphorus 370mgkg-1, calcium 43.7mgkg-1, magnesium 8.23mgkg-1, iron 9.87mgkg-1, zinc
27.7mg·kg-1。
(6) recovery of standard addition：
Claims (1)
1. a kind of method measuring micronutrient levels in blood vessel, includes the following steps：
Step 1, the preparation of test sample：
(1) the fresh blood vessel samples of 0.5g~1g are accurately weighed in glass digestion utensil, add 2~4mL top pure grade concentrated nitric acids, 90~
0.5~1mL hydrogen peroxide is added dropwise in 95 DEG C of 5~6h of Water bath, until solution is in faint yellow or white clear shape, catches up with acid to residue
1mL solution, is cooled to room temperature, and 2% nitric acid is used to be settled to 10mL as test solution；
(2) in the digestion utensil for not healing pipe sample, according to identical concentrated nitric acid in (1) and hydrogen peroxide use in identical item
It is cleared up under part, final constant volume solution is blank solution；
Step 2, Determination of trace elements：
(1) mixed standard solution series is prepared：Phosphorus, calcium, magnesium, iron, Zinc standard solution is taken to be placed in same container respectively, with 2%
Nitric acid, which dilutes, is configured to mix intermediate storing solution, and phosphorus, calcium, magnesium, iron, zinc concentration are in storing solution among the mixing
100mg·L-1；The intermediate storing solution of mixing is taken, the mixed standard solution series for preparing various concentration is diluted with 2% nitric acid, it is described mixed
The concentration of standardization solution series is respectively 0.00mgL-1、0.05mg·L-1、0.10mg·L-1、0.50mg·L-1、
1.0mg·L-1、5.0mg·L-1、10.0mg·L-1；
(2) ICP-OES determination conditions are as follows：
(3) standard curve is established：The mixed standard solution series prepared in (1) is injected separately into inductively coupled plasma spectrometry
Instrument, using response as ordinate, draws each element standard curve respectively using concentration of element as abscissa；
(4) sample measures：Blank solution and test solution are introduced into inductively-coupled plasma spectrometer respectively successively, obtained
Detection data substitutes into the standard curve that (3) are established, it is known that micronutrient levels C in blank solutionIt is emptyAnd it is micro in test solution
Constituent content CFor, micronutrient levels in sample can be obtained according to following formula calculating：
In formula, CForMicronutrient levels (mgL in test solution-1)；CIt is emptyMicronutrient levels (mgL in blank solution-1)；mSampleSample size (g).
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