CN1605869B - Blood examination method - Google Patents
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Abstract
The invention relates to an easy and simple blood checking method for determining early stage morbid states, wherein the glucose concentration, lactic acid concentration, acetyl formic acid concentration and oxidation-reduction potential in the blood erythrocyte can be measured easily, these parameters can be integrated to effectively detect miscellaneous morbid signs from the conventional blood biochemistry examination.
Description
Technical field
The present invention relates to from blood, take out the intracellular composition of regulation,, can accurately hold the blood test method of morbid state in early days by biochemical analysis.
Background technology
, took out in the past, carried out the blood test of biochemical analysis,, carried out at the medical scene of majority as one of effective health check-up inspection method of holding patient's morbid state as the composition in the blood plasma of blood liquid part.
The morbid state of wanting to hold by this blood test has many kinds, and still, for example the situation with diabetes is an example, measures patient's concentration of glucose of (in the blood plasma) in the blood on an empty stomach and after meal.Like this, the concentration of the glucose of surveying, promptly when above, the pathogenetic possibility of decidable diabetes and even glycosuria is very high up to setting for blood glucose value.In addition, it for example is the occasion of chronic respiratory organ disease or heart failure, renal failure, can measure the lactic acid concn of (in the blood plasma) in the blood, the lactic acid concn of surveying up to setting when above, can judge that the possibility that chronic respiratory organ disease or heart failure, renal failure take place is very high.
As everyone knows, people's cell be with glucose as the main energy, at night, brain, the cell of whole bodies such as nerve, haemocyte, muscle all utilizes sugar.On the other hand, liver synthesizes sugar by fat or amino acid and emits.Usually the sugared coal caving ratio of liver is consistent with the sugared utilization factor of systemic cell, so blood glucose value is below the certain value (109mg/dl).
Like this, take in by systemic cell and what effectively utilize glucose is the hormone that is called insulin.When pickuping food, saccharic forms glucose, is absorbed rapidly by small intestine, enters in the blood, thereby flows through whole body.Thus, when blood glucose value rose, moment secreted the insulin of capacity from pancreas.Like this, liver stops the release of sugar.On the other hand, become systemic cell picked-up sugar, the above-mentioned blood glucose value that rises after meal can return to the numerical value of ante cibum rapidly.Repeat the variation that this process is healthy people's a blood glucose value.
Yet, although blood glucose value rises after meal, for so-called " insulin secretion is slow; or secretory volume seldom " the people of physique, add hypomotility or excessive diet etc., when the function of insulin constantly reduces, above-mentioned glucose is difficult to take in the cell, can accumulate excessive glucose in the blood.Like this, even after meal through considerable time, blood glucose value can not reduce yet.
This situation is " hyperglycaemia " state, by general biochemical blood test method before above-mentioned, when wanting to hold this hyperglycemia state, as mentioned above, with the plasma fraction in the blood as detection bodies, carry out on an empty stomach blood glucose value (FBS) and the mensuration of blood glucose value after meal.As a result, for example, the blood glucose value in the blood plasma is more than 140mg/dl on an empty stomach, and this blood glucose value is when 200mg/dl is above after meal, and general diagnostic is diabetes (diabetes Bs).When this hyperglycemia state continued for a long time, the peculiar vascular disorders of diabetes such as retinopathy, ephrosis can appear.In addition, if blood glucose value is high more, its duration is also just long more, and retinopathy and ephrosis also just worsen more.
Yet, in recent years, use above-mentioned general biochemical blood test in the past, that is, the blood glucose value inspection in the plasma fraction, for being diagnosed as the not high patient of diabetes blood glucose value, the situation of miocardial infarction morbidity increases.That is, can confirm to give the diabetic of early stage, the light disease of main forces fully for so-called diabetes, also arteriosclerosis is also shown effect, is made progress.
For example, the blood glucose value of ante cibum belongs to healthy normal in normal range.Yet, find the situation (so-called light disease diabetes B) that each blood glucose value after meal increases unusually.Like this, though the AC Sugar value in normal range, when blood glucose value increased unusually after meal, insulin was slower than the rising of this blood glucose value, still, stimulated by hyperglycaemia, excessive secretion.Certainly, for the patient with exercise habit, above-mentioned insulin by the pancreas secretion is taken in glucose in the muscle, but for the patient who lacks motion, is taken in by adipocyte.The sugar of taking in the adipocyte becomes fat, has quickened obesity, causes hyperlipemia.Like this, hyperinsulinemia continues to cause vascular hypertension.
The result, people with the slow type physique of above-mentioned so-called insulin secretion figure, owing to meet the impaired condition of said " levis diabetes (light disease 2 types) ", " obesity is arranged slightly ", " slight hyperlipemia ", " mild hypertension ", so arteriosclerosis is made progress easily, these echo inspections by carotid wall etc. are proved.In fact, according to reports, according to the research of nearest Japan, observe when being diagnosed as the common people of diabetes in 10 years, per 10 philtrums just have the concurrent miocardial infarction of 1 people, also have the concurrent cerebral infarction of 1 people.Therefore,, before being diagnosed as diabetes, should be noted that excessive diet and lack motion that mention herein, for diabetes, requirement is paid attention in advance, the early stage necessity of grasping morbid state if the people who suffers from diabetes is arranged among the lineal relative.
Yet, above-mentioned plasma fraction with blood is as the former biochemical analysis method of detection bodies, only identical component is oozed out to the outside from the red blood cell as blood cell, presentation in blood plasma, begin to carry out analysis and judgement then, therefore in any case, judge slowly that existence may not necessarily successfully manage the above-mentioned early stage problem of grasping the requirement of morbid state.
Up to the present, the clinical red blood cell inspection of carrying out, have only mensuration, the reticulocyte of RBC number, hematocrite value, hemoglobin content, inspection of abnormal hemoglobin etc., in fact, take out intracellular fluid composition in the red blood cell, the blood test method of carrying out biochemical analysis does not easily also propose.
Disclosure of the Invention
The inventor has carried out concentrated research to carrying out the early stage blood test method of accurately holding morbid state, found that analysis arranges the endoerythrocytic composition state variation (rising of concentration of glucose of the blood cell of concentration of glucose in the blood plasma in advance, metabolism states etc.) itself is crucial for grasping early stage morbid state.Promptly, in order to grasp original early stage morbid state, as blood cell component, have the glucose of taking in the blood plasma is separated glycolytic metabolic function, know and arrange in advance at upstream side that the intracellular concentration of glucose of the red blood cell of blood glucose value (blood glucose value state) is very important in the blood plasma.Find that on the other hand if measure oxidation-reduction potential (ORP) in the blood with above-mentioned diabetes and other ill patients, during morbid state, oxidation-reduction potential is in the state of oxidation, when healthy, is in reducing condition.These all become the useful judgement parameter that can hold above-mentioned early stage morbid state, at this moment, lactic acid concn is to determining that this oxidation-reduction potential has very big influence in the above-mentioned red blood cell, the same with above-mentioned situation, with this endoerythrocytic oxidation-reduction potential, know that pyruvic acid concentration and the lactic acid concn as the saccharic metabolin is very important.The present invention is conceived to such fact, a kind of blood test method that provides is provided, it is particularly by concentration of glucose, lactic acid concn and pyruvic acid concentration in the red blood cell cell of measuring blood, and oxidation-reduction potential etc., utilize these parameters or these parameters are comprehensive respectively, suitably hold morbid state, thereby, can accurately and positively hold morbid state from the early stage light disease stage for the effective various morbid state of general blood biochemistry checking.
The present invention provides a kind of blood test method with following (a)～(h) step for achieving the above object.
(a) the mammal blood sample is centrifugal, be separated into the step of plasma fraction and haemocyte composition,
(b) remove buffycoat from this haemocyte composition, obtain erythrocytic step,
(c) with this erythrocytic step of buffering normal saline solution washing,
(d) in the red blood cell that this was cleaned, add the buffering normal saline solution, obtain the step of red blood cell suspension,
(e) this red blood cell suspension is centrifugal, remove the supernatant composition, obtain the step of red blood cell layer,
(f) in this red blood cell layer, add hypertonic pressure liquid, soak into the suspending liquid of pressing the red blood cell cellular content that oozes out, the potpourri that obtains is kept the step of sufficient time down at 25～40 ℃ for obtaining containing utilizing,
(g) this suspending liquid is centrifugal, as to collect the supernatant that contains above-mentioned red blood cell cellular content step,
(h) measure this supernatant, determine to be selected from the step of at least 1 factor in concentration of glucose, pyruvic acid concentration, lactic acid concn and the oxidation-reduction potential.
According to such formation, different with former use as the method that the plasma fraction of BECF body composition carries out blood test, by the blood cell of arranging composition state in the blood plasma in advance is endoerythrocytic composition, measure blood glucose value, or acetone acid number, lactic acid value, oxidation-reduction potential, in time, know in advance as yet not that the intracellular blood glucose value of red blood cell in the performance of blood plasma side rises and the variation of acetone acid number, lactic acid value, oxidation-reduction potential, this wants morning with showing in the blood plasma side to compare.
For this reason, to various illness situations such as diabetes, chronic respiratory organ disease, heart failure, renal failures, can make early diagnosis, and carry out corresponding treatment accurately.
The result, if utilize blood test method of the present invention, can judge correctly that then utilization was determination of plasma blood glucose value, acetone acid number, lactic acid value etc. by the outer composition of blood cell in the past, the blood test method of morbid state such as judgement diabetes can not accurately be judged the euglycemia state of value of morbid state, normal acetone acid number or low pyruvic acid state of value, normal lactic acid value or low lactic acid state of value patient's true morbid state, thereby can make early diagnosis more accurately and early treatment.
About the detailed content of blood test method of the present invention, below describe in more detail.
Description of drawings
Fig. 1 is that expression utilizes blood test method of the present invention, the figure of the typical curve of making when obtaining oxidation-reduction potential in the red blood cell.
Embodiment
Blood test method of the present invention is made of following (a)～(h) step.
(a) in the step, that venous samples can is centrifugal, be separated into plasma fraction and haemocyte composition.For example, use pack into ormal weight heparin or EDTA, gather venous blood from the examinee as the heparin tube that prevents the dicumarol that red blood cell solidifies.With the venous samples can (for example 2ml) that obtains, preferably centrifugal 5～10 minutes with 130～200 * g, especially preferably centrifugal 5 minutes with 150 * g, thus be separated into plasma fraction and haemocyte composition.The haemocyte composition is the cell component in the blood, comprises red blood cell, leucocyte, blood platelet etc.
(b) in the step, remove buffycoat, obtain red blood cell from this haemocyte composition.At this moment, as being easy to make red blood cell and the haemocyte composition beyond it, it is the separation promoter that buffycoats such as leucocyte, blood platelet separate, for example, the metrizamide that preferably in this haemocyte composition, adds 0.2ml concentration about 25%, with the potpourri that obtains, preferably centrifugal 7～12 minutes with 800～1200 * g, especially preferably centrifugal 10 minutes, be separated into the buffycoat and the red blood cell of leucocyte, blood platelet etc. with 1000 * g.Like this, if from the haemocyte composition, remove buffycoats such as leucocyte-removing, blood platelet really, then can isolate red blood cell effectively, in the step afterwards, with the composition in the high precision taking-up red blood cell, can determine endoerythrocytic glucose, lactic acid concn exactly easily.
(c) in the step, wash this red blood cell with the buffering normal saline solution.Preferably in the red blood cell of removing buffycoat, add phosphoric acid buffer normal saline solution (PBS), preferably centrifugal 5～10 minutes, especially preferably centrifugal 5 minutes, wash with 150 * g with 130～200 * g.Like this, further remove really attached to plasma fraction on the erythrocyte surface and impurity such as leucocyte, blood platelet.
(d) in the step, in this red blood cell of cleaning, add the buffering normal saline solution, obtain red blood cell suspension.Preferred adding phosphoric acid buffer normal saline solution (PBS) in this red blood cell of cleaning once more, use the automatic blood cell counter, utilize known method, make ormal weight, hematocrite value as 2ml is preferably 40～50%, is preferably 50% red blood cell suspension especially.Perhaps, also can use the automatic blood cell counter, making RBC number is 4,000,000/μ l～5,000,000/μ l, for example red blood cell suspension of 4,000,000/μ l.The red blood cell suspension that obtains is formed be difficult to destroy red blood cell, and be easy to take out the steady state (SS) of cellular content.
(e) in the step, this red blood cell suspension is centrifugal, remove supernatant, obtain the red blood cell layer.With this red blood cell suspension preferably centrifugal 5～10 minutes with 130～200 * g, especially preferably centrifugal 5 minutes with 150 * g, remove the supernatant of its phosphoric acid buffer normal saline solution (PBS), obtain the red blood cell layer.
(f) in the step, in this red blood cell layer, add hypertonic pressure liquid,, the potpourri that obtains is kept the sufficient time down at 25～40 ℃ for obtaining containing the suspending liquid of the red blood cell cellular content that oozes out owing to osmotic pressure.In order to be easy to take out the intracellular composition of red blood cell, as hypertonic pressure liquid, preferably adding concentration in this red blood cell layer is 5～10 weight %, preferred concentration is the saline solution (NaCl) of 5 weight %, making total amount is 2ml, with this potpourri preferably under 35～38 ℃, under 37 ℃, the preferred maintenance 7～15 minutes, especially preferably kept 10 minutes.As a result,, make the interior liquid composition that contains glucose and lactic component, from red blood cell, be exuded to fully in this saline solution with moisture by in the red blood cell cell and the osmotic pressure that acts between the saline solution.When temperature was lower than 25 ℃, desired one-tenth branch took out from red blood cell not exclusively, when temperature surpasses 40 ℃, had danger such as cellular content sex change.
(g) in the step, that the suspending liquid that obtains in (f) step is centrifugal, collect the supernatant that contains above-mentioned red blood cell cellular content.Preferably with this red blood cell suspension centrifugal 7～12 minutes, especially preferably centrifugal 10 minutes, collect the supernatant composition that above-mentioned red blood cell cellular content oozes out with 1750 * g with 1500～2000 * g.
(h) in the step, measure this supernatant, determine to be selected from least one parameter in concentration of glucose, pyruvic acid concentration, lactic acid concn (L-lactic acid concn (mg/dl)) and the oxidation-reduction potential.
Biochemical automatic analysing apparatus be can utilize,, concentration of glucose in this supernatant (D-concentration of glucose), pyruvic acid concentration, lactic acid concn (L-lactic acid concn) measured with known proper method.In (d) step, when making the 2ml hematocrite value and be 50% red blood cell suspension, concentration of glucose, pyruvic acid concentration, lactic acid concn (mg/dl) are that each measured value multiply by 2 value in the red blood cell of above-mentioned venous samples can.On the other hand, in (d) step, utilize the automatic blood cell counter, make RBC number when being the ormal weight red blood cell suspension of setting, concentration of glucose in the red blood cell of above-mentioned venous samples can (D-concentration of glucose (mg/dl)), pyruvic acid concentration (mg/dl), lactic acid concn (L-lactic acid concn (mg/dl) can be easily by each measured value, calculate with mean corpuscular volume, RBC number and the red blood cell suspension liquid measure of automatic blood cell counter while instrumentation.
In addition, utilize oxidation-reduction potentiometer (ORP meter), can measure the oxidation-reduction potential (Rc-ORP) of this supernatant with suitable known method.In above-mentioned experimentation, following the making of typical curve when to use 5% saline solution to make hematocrite value be 50% red blood cell suspension.Promptly, think that at extracellular fluid be on the extended line of plus or minus direction of oxidation-reduction potential of serum, the oxidation-reduction potential that has intracellular fluid, thereby it is as shown in table 1, the serum that use is gathered from same examinee, make typical curve shown in Figure 1,, obtain the intracellular oxidation-reduction potential of final red blood cell (Rc-ORP) according to the oxidation-reduction potential (ORP) of using above-mentioned oxidation-reduction potentiometer actual measurement.
Table 1
| The A point | The B point | The | |
| X | |||
| 1. the ORP of |
2. the ORP after |
3. the ORP after |
| Y | With the ORP after 1. being 2 times with the dilution of 5% saline solution | With the ORP after 2. being 2 times with the dilution of 5% saline solution | With the ORP after 3. being 2 times with the dilution of 5% saline solution |
At first, use the heparin tube of pack into ormal weight blood coagulation accelerator and serum separating agent, the venous blood that ormal weight is gathered from the examinee, 4ml for example with for example 3000rpm centrifugal about 10 minutes, can obtain serum.Then obtain the oxidation-reduction potential of serum respectively and be the oxidation-reduction potential of this serum of 2 times with 5% saline solution dilution, with the point determined thus as A point in the diagram.Then, by with former serum be with phosphoric acid buffer normal saline solution (PBS) dilution 2 times serum oxidation-reduction potential and will use phosphoric acid buffer normal saline solution (PBS) dilution to be that 2 times serum dilutes with 5% saline solution again be that the point that obtains of 2 times the oxidation-reduction potential of serum is as illustrating the B point.Equally, the C point serve as reasons with former serum with phosphoric acid buffer normal saline solution (PBS) dilution be 4 times serum oxidation-reduction potential and will be that 4 times serum dilutes with 5% saline solution again with phosphoric acid buffer normal saline solution (PBS) dilution be the point that 2 times the oxidation-reduction potential of serum is obtained.A, B, C each point are connected, draw extended line to the plus or minus direction, with it as typical curve.Then, apply mechanically the measured value of oxidation-reduction potential of the mixed liquor of intracellular fluid and above-mentioned 5% saline solution on the Y-axis of typical curve formerly (coordinate axis of the oxidation-reduction potential that serum is obtained with 5% saline solution dilution), read the oxidation-reduction potential of the intracellular fluid of inferring by the coordinate of X-axis.
In addition, in above-mentioned (h) step, use above-mentioned oxidation-reduction potentiometer (ORP meter) when obtaining oxidation-reduction potential (ORP), measured value is added the current potential of comparison electrode.This is the current potential of electrode relatively, and the temperature of supernatant composition is determined according to this moment, and for example, this temperature is between 0-60 ℃, and is as shown in table 2.
Table 2
Further specify the present invention by following examples.
By following (a)～(i) step,, measure concentration of glucose (D-concentration of glucose) in the red blood cell cell of any time to 52 of 37 of insulin dependent/non-dependent diabetics (NIDDM patient) and healthy normal persons.
Step (a):, be separated into plasma fraction and haemocyte composition with 2ml venous samples can centrifugal 5 minutes with 150 * g.
Step (b): after in this haemocyte composition, adding 0.2ml concentration and be 25% metrizamide, with 1000 * g centrifugal 10 minutes, be separated into buffycoat and red blood cells such as leucocyte, blood platelet.
Step (c): after in this red blood cell, adding phosphoric acid buffer normal saline solution (PBS), with 150 * g centrifugal 5 minutes, wash.
Step (d): add phosphoric acid buffer normal saline solution (PBS) in this red blood cell of cleaning, use the automatic blood cell counter, making 2ml hematocrite value is 50% red blood cell suspension.
Step (e): with this red blood cell suspension centrifugal 5 minutes, remove the supernatant of this phosphoric acid buffer normal saline solution (PBS), obtain the red blood cell layer with 150 * g.
Step (f): add the saline solution (NaCl) of 5 weight % in this red blood cell layer, making total amount is 2ml, and this potpourri was kept 10 minutes down at 37 ℃, obtains suspending liquid.
Step (g):, obtain the supernatant composition that the intracellular composition of red blood cell oozes out with suspending liquid centrifugal 10 minutes with 1750 * g.
Step (h): measure this supernatant, determine concentration of glucose.
In addition, adopt and former the same method, measure the blood glucose value in the blood plasma of blood respectively.
The result, insulin dependent/non-dependent diabetic's occasion, when the blood glucose value in the blood plasma is very high, the intracellular concentration of glucose of red blood cell (D-concentration of glucose) is also very high, blood glucose value is 350mg/dl, concentration of glucose (D-concentration of glucose) also reaches 50mg/dl in the red blood cell cell,, confirms to exist positive correlativity between the two that is.
On the other hand, normal health adult's occasion, when blood glucose value reached 150mg/dl, concentration of glucose (D-concentration of glucose) was 10mg/dl to the maximum in the red blood cell cell, confirmed still to exist positive correlativity.
From these insulin dependent/non-dependent patients and normal health adult's total data as can be seen, exist significantly positive correlativity between the concentration of glucose in blood glucose value in blood plasma and the red blood cell cell (D-concentration of glucose).
With the method identical, carrying out property angiodystrophia patient (4 years old, the male sex) is carried out blood test with embodiment 1.The results are shown in table 3. Measure 1 and 2 be respectively carry out before the negative ion irradiation treatment and after measurement result.
Table 3
| |
|
|
| The interior pyruvic acid internal pH of lactic acid cell in the ORP cell in the cell | 248mV 10.6mg/dl 0.68mg/dl 7.13 | 221mV 18.4mg/dl 1.2mg/dl 7.24 |
| Pyruvic acid blood pH in the lactacidemia in the blood ORP blood | 136mV 16.8mg/dl 0.8mg/dl 7.39 | 121mV 18.2mg/dl 0.9mg/dl 7.44 |
| CPK myoglobins LDH aldolase | 29475U/l 1700ng/ml 1550IU/l 169.2IU/l | 15854U/l 840ng/ml 1177IU/l 96.2IU/l |
In this case, measure at 1 o'clock, ORP high (248mV) in the cell thinks that intracellular oxidation is remarkable.For this reason, since the oxidation of cell membrane, Na
+/ H
+The driving force of passage reduces, and intracellular pH also reduces (7.13).Usually, although lactic acid is higher slightly than lactic acid in the blood in the cell, lactic acid is 10.6 in the cell, 16.8 quite low than lactic acid in the blood.Do not have the TCA loop in the red blood cell, only separate sugar system, therefore, after sugar decomposition becomes pyruvic acid, be varied to lactic acid and finish.Lactic acid means that than low this fact of lactic acid in the blood intracellular metabolism has reduced in the cell.That is, internal pH reduces, and ORP raises, so intracellular acidity, oxidation are hyperfunction, metabolism reduces.At this moment, CPK, myoglobins, LDH, the aldolase of expression muscle inflammation have reached remarkable high value, patient's ill extreme difference.On the other hand, measure at 2 o'clock, ORP hangs down to 221 in the cell, and internal pH also approaches normally (7.24), and not significant acid, oxidation in the cell thinks metabolic good.In fact, lactic acid and pyruvic acid are respectively 18.4 and 1.2 in the cell, present than the high slightly trend of these values in the blood.In any case, these measured values are than normal value height, and cell presents acidity, so need to improve.Therefore, the inside and outside organic acid concentration of research cell is perhaps measured pH and ORP, can hold the generation or the patient's condition of disease early.As a result, for measuring 2, the value of CPK, myoglobins, LDH, aldolase has all had remarkable improvement than measuring 1.In fact, this patient stands and walking disorder owing to muscular atrophy causes, but in the stage of measuring 2, symptom is improved significantly.
Embodiment 3
With the method identical, diabetic (70 years old, the male sex) is carried out blood test with embodiment 2.The results are shown in table 4. Measure 1 and 2 be respectively carry out before the negative ion irradiation treatment and after measurement result.
Table 4
| |
|
|
| The interior pyruvic acid internal pH of lactic acid cell in the ORP cell in the cell | 229mV 17.5mg/dl 0.61mg/dl 7.11 | 250mV 23.8mg/dl 0.4mg/dl 7.28 |
| Pyruvic acid blood pH in the lactacidemia in the blood ORP blood | 132mV 15.9mg/dl 0.5mg/dl 7.33 | 121mV 18.2mg/dl 0.6mg/dl 7.37 |
| Blood glucose value | 380mg/dl | 309mg/dl |
Measuring in 1, lactic acid is 17.5 in the cell, and is higher slightly than 15.9 in the blood, because its measured value is just over normal value, at first sight the metabolism of cell is so not poor.And ORP is 229 also in the cell, thinks that not have oxidation hyperfunction.Yet internal pH is 7.11, significantly reduces, so think that intracellular glycometabolism is in halted state.On the other hand, measuring in 2, because lactic acid very high (23.8) in the cell, ORP is also up to 250, so predict that because of cellular oxidation, metabolism worsens.Yet, because the internal pH optimum, so think and carry out glycometabolism in this stage.In fact, blood sugar is 309, compares with 380 of mensuration 1, has obtained good improvement.
According to the organic acid inside and outside the cell, pH and ORP, can prediction the possibility of morbidity and the quality of the patient's condition.According to the data result of present collection, think that the process order the when morbidity or the patient's condition worsen is:
1. with cell inside and outside the balance of organic acid measured value worsen
2. with cell inside and outside the balance of measured value of ORP worsen
3. with cell inside and outside the balance of pH measured value worsen
Reference value is as follows.
Below the interior ORP 235mV of cell
Lactic acid 8～16mg/dl in the cell
Pyruvic acid 0.5～1.2mg/dl in the cell
Internal pH 7.24～7.30
Below the blood ORP 130mV
Lactic acid 6～14mg/dl in the blood
Pyruvic acid 0.3～0.9mg/dl in the blood
Blood pH 7.36～7.42
Claims (3)
1. a blood test method is characterized in that may further comprise the steps, that is,
(a) with the centrifugal force of mammal blood sample with 130～200 * g, centrifugal 5～10 minutes, be separated into the step of plasma fraction and haemocyte composition,
(b) in this haemocyte composition, add precipitation agent,, remove buffycoat, obtain erythrocytic step from this haemocyte composition with the potpourri that obtains centrifugal 7～12 minutes with 800～1200 * g,
(c) in this red blood cell, add the phosphoric acid buffer normal saline solution, with the potpourri that obtains centrifugal 5～10 minutes with 130～200 * g, with this erythrocytic step of buffering normal saline solution washing,
(d) add the buffering normal saline solution in the red blood cell that this was cleaned, with the potpourri that obtains automatic blood cell counter, the hematocrite value that is made into ormal weight is the step that 40～50% red blood cell suspension obtains red blood cell suspension,
(e) this red blood cell suspension is centrifugal with centrifugal 5～10 minutes of 130～200 * g, remove the supernatant composition, obtain the step of red blood cell layer,
(f) saline solution of adding 5～10 weight % in this red blood cell layer, making total amount is ormal weight, and this potpourri was kept 7～15 minutes down at 35～38 ℃, obtains containing the step that the suspending liquid of pressing the red blood cell cellular content that oozes out is soaked in utilization,
(g) this suspending liquid is centrifugal with centrifugal 7～12 minutes of 1500～2000 * g, collect the step that contains the supernatant that above-mentioned red blood cell cellular content oozes out,
(h) measure this supernatant, determine to be selected from the step of at least 1 factor in concentration of glucose, pyruvic acid concentration, lactic acid concn and the oxidation-reduction potential.
2. according to the blood test method of claim 1 record, it is characterized in that, utilize biochemical automatic analysing apparatus, measure concentration of glucose, pyruvic acid concentration, lactic acid concn in this supernatant.
3. according to the blood test method of claim 1 record, it is characterized in that, utilize oxidation-reduction potentiometer to measure oxidation-reduction potential in this supernatant.
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| EP2885636B1 (en) * | 2012-10-23 | 2018-02-14 | Aytu BioScience, Inc. | Methods and systems for measuring oxidation-reduction potential of a biological sample |
| CN103808774B (en) * | 2014-02-26 | 2016-01-13 | 高磊 | Mean corpuscular concentration of glucose |
| CN107407650B (en) * | 2015-03-31 | 2020-09-11 | 索尼公司 | Electrical characteristic measuring device, electrical characteristic measuring method, blood condition analyzing system, and electrical characteristic measuring program for computerized method |
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| EP0342398A1 (en) * | 1988-05-14 | 1989-11-23 | Dimitrios Dr. Giannitsis | Process for determining antibodies against antigens specific for erythrocytes |
| CN1171553A (en) * | 1996-04-04 | 1998-01-28 | 生命扫描有限公司 | Reagent test strip for blood glucose determination |
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| EP0342398A1 (en) * | 1988-05-14 | 1989-11-23 | Dimitrios Dr. Giannitsis | Process for determining antibodies against antigens specific for erythrocytes |
| CN1171553A (en) * | 1996-04-04 | 1998-01-28 | 生命扫描有限公司 | Reagent test strip for blood glucose determination |
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