CN1122837C - Inorganic ion ultramicro-detecting technology and method using multifunctional biochemical enzyme labelling analyzer - Google Patents
Inorganic ion ultramicro-detecting technology and method using multifunctional biochemical enzyme labelling analyzer Download PDFInfo
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- CN1122837C CN1122837C CN 00134389 CN00134389A CN1122837C CN 1122837 C CN1122837 C CN 1122837C CN 00134389 CN00134389 CN 00134389 CN 00134389 A CN00134389 A CN 00134389A CN 1122837 C CN1122837 C CN 1122837C
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Abstract
The present invention provides a method using the existing porous plate and directly making testing operation in pores. In the method, polyethylene plastic tubes are made into micro blood-taking plastic tubes with different specifications, and a quantitative sample-adding (suction-shifting) device is used for attaining a goal without the need of taking blood from a vein, so that standard products can reach the standard with known contents every time after multiple standard products with known contents are simultaneously tested with a tested sample in an inorganic ion testing item so as to enhance the accuracy of the result of the tested sample. A test report can be printed by combining the test of a correspondingly matched multifunctional biochemical enzyme labeling analyzer.
Description
Technical field
The invention belongs to medical science electrolyte check field.Be a kind of method specifically with multifunctional biochemical enzyme label analyzer test inorganic ions ultramicron detection technique.
Background technology
The inorganic ions check is one of important component part of medical test.The inorganic ions analysis towards trace, fast, accurately, easily direction to develop be the inexorable trend of current domestic and international medical test.
At present, both at home and abroad the advanced person's who produces full-automatic potassium, sodium, chlorine, calcium analyser, full-automatic electrolysis matter analyser how trace, fast, accurately, automaticity is high, but all do not break away from electrode, and electrode life is shorter, changes one time the cost height.In view of China's present situation, owing to be subjected to limitations of all sorts of conditions such as economy, maintenance, reagent, so far still fail generally to promote advanced full-automatic electrolysis matter analyser, what most laboratory was still used is flame photometric analysis method and colourimetry commonly used, and this two method all exists unstable result, poor repeatability, and all needs the artificial deficiency of calculating.The flame photometric analysis method has deionized water, sample to deposit the shortcoming that the high and used fuel gasoline of requirement, liquefied petroleum gas (LPG) etc. have certain danger.Colourimetry commonly used has colorimetric test speed hand-manipulated to wait shortcoming slowly.Can realize not only quick but also colorimetric analysis accurately
Technical method, the practical significance that has bigger technical superiority and use.
The objective of the invention is to avoid above-mentioned the deficiencies in the prior art part and adopt existing microwell plate replacing test tubes and cuvette, and in the hole, directly carry out test operation, reach the standard items that add a plurality of known content numbers, a kind of method that the content that provides after tested is whether consistent with multifunctional biochemical enzyme label analyzer test inorganic ions ultramicron detection technique.
Summary of the invention
Purpose of the present invention can reach by following measure:
A kind of method with multifunctional biochemical enzyme label analyzer test inorganic ions ultramicron detection technique is characterized in that:
A. press tip routine blood test acquisition method: with trace blood plastic tube upper port contact drop of blood, lower in the lower end, the limit is gently squeezed the limit and is entered blood scorification lower end to the requirement, clutch sealing or open-ended with tweezers, the trace blood plastic tube is converted in the syringe needle plastic jacket of " V " type insertion disposable syringe, stick the adhesive sticker sequence number on syringe needle plastic casing or blood sampling plastic tube, centrifuging serum is standby;
B. get one of clean 8 * 12 standards, 96 microwell plate;
C. adopt the micro quantitative determination sample injector, get standard items and sample, quantitatively add in each hole of microwell plate;
D. use micro-adjustable pipette,, quantitatively add in above-mentioned each hole by required reagent dosage;
E. microwell plate is put room temperature and reached the required reaction time of test;
F. microwell plate is put in the multifunctional biochemical enzyme label analyzer measuring chamber, in 90s, printed 1～90 part of result's report simultaneously.
Purpose of the present invention can also reach by following measure:
The present invention adopts l.2 μ μ μ μ μ μ l micro quantitative determination sample injector l.25 l.20 l.10 l.5 l.3 of 1 μ, get standard items in 2-4 hole and sample quantitatively add in each hole of microwell plate.
The present invention adopts 50-250 μ l, 100-500 μ l trace adjustable pipette.
The present invention adopts the vinyl tube of internal diameter 2-3mm to make the trace blood plastic tube of 5-7cm specification.
The present invention's method compared to existing technology has following advantage:
1. the inorganic ions method for measuring has a lot, and the most frequently used is colourimetry.Method is reliable, but poor repeatability tends to occur different results with a sample repeated detection.Tracing it to its cause, except sampling, application of sample, operation factors, mainly is the precision and the insufficient sensitivity of detecting instrument.And the present invention adopts a whole set of to adapt to multifunctional biochemical enzyme label analyzer and corresponding techniques method that inorganic ions ultramicron detects, has not only improved the precision of instrument and sensitivity but also can make with a part sample, is consistent substantially with crowd sample test result.
2. colourimetry and flame photometric analysis method all need manually be carried out colorimetric, speed is slow, exist between the colorimetric time period error to the result.And with art methods in sample in enormous quantities detects, avoided directly influencing the accuracy of measurement result because of the standard items and the overlong time of sample front and back colorimetric.
3. fast with art methods colorimetric analysis speed.90 duplicate samples only can test printing go out result's report with 90s.
4. various consumptive materials are few.From getting blood, need not syringe, test tube, suction pipe, cuvette to sending report.Reduce cleaning, saved manpower, improved work efficiency.
5. in view of the advanced person's of domestic and international production full-automatic electrolysis matter analyser, though trace, fast, accurately, automaticity is high, measures the easy bubble in chamber, pipeline easily stops up, and more do not break away from electrode, and electrode life is shorter, changes one time the cost height.What China's most laboratory was still used at present is flame photometric analysis method and colourimetry commonly used, and flame photometer has certain danger because of used fuel gasoline, liquefied petroleum gas (LPG) etc. again.And the present invention adopts a whole set of to adapt to multifunctional biochemical enzyme label analyzer and corresponding techniques method that the inorganic ions ultramicron detects, be with the standard items of 2-4 known content number with sample after tested after, all can reach the standard of known content number at every turn, improve tested sample result's accuracy.
Embodiment
Embodiment 1
Serum calcium measuring (neighbour-cresolphthalein complexon direct colo(u)rimetry)
1. tip blood is gathered:
(1) the blood sampling site adult is advisable with the left hand third finger, and infant below half years old is usually with thumb or heel blood sampling.
(2) massage blood sampling site gently, make it congested naturally, with 75% ethanol cotton balls sterilize local skin, wait to do.
(3) tightly pinch thorn blood position, get blood with the puncture of sterile blood sampling pin.Action should be rapid, and the about 2-3mm of the degree of depth can flow out with squeezes blood a little and to be advisable.
(4) get vinyl tube (trace blood plastic tube) the upper port contact drop of blood of the long 5-7cm specification of internal diameter 2-3mm, lower in the lower end, the limit is gently squeezed the limit and is advanced blood, adopt the 1 scorification lower end, back of bleeding approximately, clutch sealing with tweezers, or open-ended, the trace blood plastic tube is converted in the syringe needle plastic jacket of " V " type insertion disposable syringe, stick the adhesive sticker sequence number on syringe needle plastic casing or blood sampling plastic tube, centrifuging serum is standby.
2. principle:
Calcium ion in the sample under alkali condition with neighbour-cresolphthalein complexon complexing, generate the aubergine complex compound, special absorption peak is arranged under the 580nm wavelength, the depth of color is directly proportional with the concentration of calcium.And the magnesium ion in the adding oxine chelating sample is obtained the content of calcium to eliminate interference by colorimetric estimation.
3. reagent:
(1), neighbour-cresolphthalein complexon developer: take by weighing oxine 500mg, put in the beaker, add concentrated hydrochloric acid 5ml, make its dissolving and change in the 500ml volumetric flask, add neighbour-cresolphthalein complexon 25mg again, after treating to dissolve fully, add Tri-tonX-100 1ml, mixing adds deionized water then to scale.Put in the polyethylene bottle and preserve.
(2), 1mol/L AMP alkaline buffer: take by weighing 2-amino-2-methyl-1-propanol 89.14g, put in the 1L volumetric flask, add the 500ml deionized water dissolving, add to scale after treating to dissolve fully.Put room temperature preservation in the polyethylene bottle.
(3), colour developing uses liquid: during application, according to what of sample with above-mentioned two liquid mixed in equal amounts.
(4), standard calcium liquid: 2.5mmol/L
4. application of sample: get according to the form below application of sample of clean 8 * 12 standards, 96 microwell plates:
Admixture (μ l) blank well gauge orifice is measured the hole
Distilled water 5
Titer (2.5mmol/L) 5
Serum (slurry) 5
Liquid 300 300 300 is used in colour developing
Mixing was put room temperature 5 minutes.
5. test: will test microwell plate and put in the multifunctional biochemical enzyme label analyzer measuring chamber, with the single wavelength of 580nm, establish blank well and gauge orifice position, confirm the Au/As*C formula, the product unit content sets up standard: the standard calcium product promptly are provided with C=OO2.5 for 2.5mmol/L, start to measure result's report of printing each hole serum calcium levels.
6. blood calcium determination as a result: neighbour-cresolphthalein complexon-colourimetry reagent: Beijing northization fine chemicals Ltd product batch number: 000110 wavelength mode: single wavelength measurement wavelength: 580nm titer (product): 2.5mmol/L report manner: 1, blood calcium test result 2, the absorbance test result is used formula: Au/As*C standard items unit content is provided with: two kinds of report manners of C=OO25 (2.5mmol/L) are as follows: 1, blood calcium determination result: A capable 01,02,03 hole is blank " O " hole of transferring
04,05,06,07 hole is the standard items measurement result,
All the other measure the hole result for sample.2, absorbance test result: with corresponding each the hole absorbance of results of biochemical
Blood calcium determination is plate number as a result: 01 date: 00-07-02 time: 09:30 blank well: mode: single wavelength identification code: 01 wavelength: 580 humidity: temperature:
Negative control hole value (N) :+0.797
Formula: Au/As*C
01 02 03 04 05 06 07 08 09 10 11 12A +000.1 -000.0 -000.0 +002.5 +002.6 +002.5 +002.5 +002.2 +002.1 +001.7 +001.8 +001.8
Absorbance test result plate number: 01 date: 00-07-02 time: 09:31 blank well: mode: single wavelength identification code: 01 wavelength: 580 humidity: temperature:
Blank well value :+0.258
01 02 03 04 05 06 07 08 09 10 11 12
+0.036 -0.012 -0.023 +0.788 +0.824 +0.789 +0.786 +0.704 +0.673 +0.547 +0.563 +0.563
Embodiment 2 chloride determinations (mercuric thiocyanate colourimetry)
1. tip blood is gathered with embodiment 1.
2. principle: chlorion in the determinand and mercuric thiocyanate effect, generate the mercuric chloride that is difficult to dissociate, and discharge the thiocyanate ion of respective amount, this ion combines the orange-red ferric rhodanate of generation with ferric ion in the reagent, and its color and luster is directly proportional with the content of chlorine.
3. reagent:
(1) saturated mercuric thiocyanate solution: take by weighing mercuric thiocyanate 2.0g, be dissolved in the 1L distilled water, put room temperature 48 hours, and often shake, get supernatant and use.
(2) mercuric nitrate solution: take by weighing mercuric nitrate 6.0g, use the 50ml dissolved in distilled water, add the 1ml red fuming nitric acid (RFNA) and be diluted to 100ml.
(3) liquid is used in colour developing: take by weighing ferric nitrate [Fe (No3) 39H2O] 13g, add the about 400ml dissolving of water, add 1.5ml red fuming nitric acid (RFNA), the saturated mercuric thiocyanate solution of 500ml and 5.0ml mercuric nitrate solution again, be diluted with water to 1000ml at last.Deposit with plastic bottle, put room temperature preservation.
(4) chloride titer: 100mmol/L.Blank reagent: take by weighing ferric nitrate 13g, be dissolved in the 400ml distilled water, add red fuming nitric acid (RFNA) 1.5ml, be diluted to 1000ml again.
4. application of sample: get according to the form below application of sample of clean 8 * 12 standards, 96 microwell plates:
Admixture (μ l) blank well gauge orifice is measured the hole
Blank reagent 5
Titer (100mmol/L) 5
Serum (slurry) 5
Liquid 300 300 300 is used in colour developing
Mixing was put room temperature 10 minutes.5. test: will test microwell plate and put in the multifunctional biochemical enzyme label analyzer measuring chamber, with the 490nm dual wavelength, establish blank well and gauge orifice position, confirm the Au/As*C formula, the product unit content sets up standard, for example: the chlorine standard items are that 100mmol/L promptly is provided with C=100.0, start to measure result's report of printing each hole chlorinity.
6. result:
Chloride determination: mercuric thiocyanate colourimetry
Reagent: Beijing northization fine chemicals Ltd produces
Lot number: 000202
Wavelength mode: dual wavelength
Measure wavelength: 490nm
Titer (product); 100mmol/L
Report manner: 1, chloride determination result 2, absorbance test result
Use formula: Au/As*C
The standard items unit content is provided with: C=100.0 (100mmol/L)
Two kinds of report manners are as follows;
1, capable 01,02 hole of chloride determination result: A is blank " O " hole of transferring, 03,
04,05 hole is the standard items measurement result, and all the other are mark
This mensuration hole result.
2, absorbance test result: with corresponding each the hole absorbance of chlorine measurement result
Chloride determination is plate number as a result: 01 date: 00-07-11 time: 10:28 blank well: mode: dual wavelength code: 01 wavelength: 490 humidity: temperature:
Negative control hole value (N) :+1.086
Formula: Au/As*C
01 02 03 04 05 06 07 08 09 10 11 12A +000.2 -000.2 +099.5 +100.6 +099.8 +030.3 +103.2 +103.2 +099.7 +033.4 +094.9 +090.0B +110.5 +082.6 +077.9 +048.8 +094.7 +091.1 +140.2 +101.0 +095.9 +033.9 +110.0 +092.8
Absorbance test result plate number: 01 date: 00-07-11 time: 10:29 blank well: mode: dual wavelength code: 01 wavelength: 490 humidity: temperature:
Blank well value :+0.536
01 02 03 04 05 06 07 08 09 10 11 12A +0.002 -0.002 +1.081 +1.093 +1.085 +0.329 +1.122 +1.122 +1.083 +0.363 +0.923 +0.979B +1.201 +0.598 +0.847 +0.530 +0.029 +0.990 +1.524 +1.098 +1.043 +0.368 +1.195 +1.009
Claims (3)
1, a kind of method with multifunctional biochemical enzyme label analyzer test inorganic ions ultramicron detection technique, this method comprises: get one of clean 8 * 12 standards, 96 microwell plate; Adopt the micro quantitative determination sample injector, get standard items and sample, quantitatively add in each hole of microwell plate; Use micro-adjustable pipette,, quantitatively add in above-mentioned each hole by required reagent dosage; Microwell plate is put room temperature reach the required reaction time of test; Microwell plate is put in the multifunctional biochemical enzyme label analyzer measuring chamber, in 90s, printed 1～90 part of result's report simultaneously; It is characterized in that: at first press tip routine blood test acquisition method, then with vinyon trace blood pipe upper port contact drop of blood, lower in the lower end, the limit is gently squeezed the limit and is advanced blood to requirement, the trace blood plastic tube is converted in the syringe needle plastic jacket of " V " type insertion disposable syringe, stick the adhesive sticker sequence number on syringe needle plastic casing or blood sampling plastic tube, centrifuging serum is standby.
2, the method with multifunctional biochemical enzyme label analyzer test inorganic ions ultramicron detection technique according to claim 1, it is characterized in that: adopt 1 μ l, 2 μ l, 3 μ l, 5 μ l micro quantitative determination sample injectors, get standard items and in 2-4 hole, quantitatively add in each hole of microwell plate with sample.
3, the method with multifunctional biochemical enzyme label analyzer test inorganic ions ultramicron detection technique according to claim 1, it is characterized in that: the vinyl tube of employing internal diameter 2-3mm is made the trace blood plastic tube of 5-7cm specification.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 00134389 CN1122837C (en) | 2000-12-12 | 2000-12-12 | Inorganic ion ultramicro-detecting technology and method using multifunctional biochemical enzyme labelling analyzer |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 00134389 CN1122837C (en) | 2000-12-12 | 2000-12-12 | Inorganic ion ultramicro-detecting technology and method using multifunctional biochemical enzyme labelling analyzer |
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| Publication Number | Publication Date |
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| CN1299969A CN1299969A (en) | 2001-06-20 |
| CN1122837C true CN1122837C (en) | 2003-10-01 |
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Granted publication date: 20031001 Termination date: 20101212 |