WO2001081930A2 - Reactif et procede d"analyse de «fer serique » dans le plasma - Google Patents

Reactif et procede d"analyse de «fer serique » dans le plasma Download PDF

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Publication number
WO2001081930A2
WO2001081930A2 PCT/US2001/012927 US0112927W WO0181930A2 WO 2001081930 A2 WO2001081930 A2 WO 2001081930A2 US 0112927 W US0112927 W US 0112927W WO 0181930 A2 WO0181930 A2 WO 0181930A2
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iron
buffer
composition
indicator
serum
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PCT/US2001/012927
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English (en)
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WO2001081930A3 (fr
Inventor
Yen Yue
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Dade Behring Inc.
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Publication of WO2001081930A2 publication Critical patent/WO2001081930A2/fr
Publication of WO2001081930A3 publication Critical patent/WO2001081930A3/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/90—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving iron binding capacity of blood

Definitions

  • This invention relates to a composition and process for measuring iron.
  • the invention also provides a method for the determination of the total iron binding capacity in either serum or plasma.
  • Iron exists in the body mainly as hemoglobin in red blood cells and in the liver, the spleen, the bone marrow, and the like. About two-thirds of the body's iron is found in the hemoglobin ( ⁇ 2.5g).
  • the total weight of iron in a human body is about 4 grams.
  • the body loses about one to 2 milligrams of iron per day. Therefore, the balance can be maintained by absorbing about one to 2 milligrams of iron from food every day.
  • the average American diet provides 10 to 15 mg of iron daily. Normally, the body is under very tight control to absorb 1 to 2 mg of iron from food to maintain the balance. If too little iron is taken in, an anemic condition may occur. If too much is taken in, an iron overload condition may occur.
  • the Approximate Iron distribution in the body is as follows: Hemoglobin: -2.5 g
  • Myoglobin -130 mg Labile pool: ⁇ 80 mg Tissue: ⁇ 8 mg (as co-enzyme)
  • Iron storage as ferritin and Hemosiderrin: -800 mg for men and 0 to 200 mg for women.
  • Transferrin is the principal plasma protein for transport of iron. Its concentration correlates with the total iron-binding capacity (TIBC) of serum. Transferrin is a betal globulin of 75,000-80,000 Dalton. The iron in serum/plasma is physiologically bound to transferrin. The amount of iron bound to transferrin is about four milligrams. Transferrin has the capacity to bind two iron atoms per molecule in the ferric (trivalent) form. The main function of transferrin is to transport and transfer iron from the mucous membrane of the small intestine to the red blood cell precursors in bone marrow, where it is incorporated into hemoglobin.
  • TIBC total iron-binding capacity
  • the amount of iron bound to transferrin is commonly referred to as the serum iron.
  • Serum is usually used for the iron assay for reasons of technical capability.
  • the amount of iron in serum/plasma (not including hemoglobin) is about 100 micrograms per 100 milliliters of blood.
  • the amount of transferrin not bound to iron is referred to as the unsaturated iron-binding capacity (UIBC) in serum.
  • the UIBC is nonnally about 200 microgram per 100 milliliter of blood.
  • the total amount of transferrin present in blood represents the total iron binding capacity. Normally the total amount of transferrin is about 300 micrograms per 100 milliliters of blood.
  • the unsaturated iron binding capacity (UIBC) can be calculated as the difference between the total iron binding capacity (TIBC) and the serum iron.
  • the unsaturated iron binding capacity relate to the metabolism of hemoglobin and its measurement is indispensable for a differential diagnosis of anemic persons.
  • the tests also used to monitoring pregnancy, Hepatitis, acute inflammation (respiratory), abscess immunization, myocardial infarction and chronic inflammation or malignancy.
  • the amount of iron measured in serum can be done by known methods.
  • serum iron is assayed by adding a serum sample to a reagent buffered at an acid pH. At this acid pH, ferric ion dissociates from transferrin.
  • the reagent includes a reducing agent, which aids in the dissociation process and reduces ferric ion to ferrous ion.
  • a chromogenic reagent is then added and the chromogen complexes with ferrous iron to form a colored complex. The colored complex is measured spectrophotometrically.
  • There are also known methods to determine the unsaturated binding capacity of serum There are also known methods to determine the unsaturated binding capacity of serum.
  • the unsaturated iron binding capacity is the difference between the TIBC and the serum iron and is normally about 200 microgram per 100 milliliter of blood.
  • UIBC represent the free iron-binding sites available. It can be measured by adding known amount of excess iron to the serum sample at neutral pH. Added iron binds to unbound transferrin. The amount of iron remaining unbound after sample addition is measured photometrically and the difference in the original known amount less the remaining amount represents the UIBC.
  • Another method to determine UIBC comprises adding an alkaline buffer solution to serum, adding a known amount of iron to the mixture to bind all unbound transferrin to the added iron, and colorimetrically determining the residual iron to measure the unsaturated iron-binding capacity [the Schade method: Proc. Soc. Exp. Biol. & Med., 87, 443-448 (1954)].
  • the color-forming reaction rate is maximum at about pH 5.0 in the case of using bathophenanthroline as a color-forming reagent.
  • the color-forming rate is slowed down when the pH is either higher or lower than that pH (about 5.0).
  • bathophenanthroline is used as a color-forming reagent in the Schade method
  • the length of the color- formation in the Schade method makes it difficult to use an auto analyzer, although the Schade method is a very simple method without using centrifugal separation, removal of proteins and complicated procedures.
  • U.S. 5,420,008 Another method to measure iron and unsaturated iron binding capacity is disclosed in U.S. 5,420,008.
  • the methods require the use of the enzyme aconitase.
  • a pH between 5-9 and optimally 6-8 is maintained to bind iron to aconitase.
  • the buffers to maintain the basic pH include PIPES buffer, tris buffer, Triethylnolamine buffer, glycine buffer, GTA buffer, and phosphate buffer.
  • the method to measure unsaturated iron binding capacity requires a stable, alkaline, excess iron solution of known concentration to fully saturate the transferrin with iron.
  • a somewhat stable bivalent iron compound (ammonium iron (II) sulfate or ferrous chloride) rather than a trivalent iron compound, is used in a Tris buffer solution in the co-presence of a reducing reagent such as ascorbic acid for stabilization.
  • a reducing reagent such as ascorbic acid for stabilization.
  • the unbound iron is measured by binding it to aconitase.
  • the TIBC is most clinicians' preferred method, the UIBC has been easier to automate than the TIBC.
  • the total iron binding capacity (TIBC) is the maximum amount of iron that can be bound. The iron binding capacity of serum is accounted for almost entirely by transferrin.
  • TIBC can be measured by adding excess iron to saturate all the transferrin iron binding sites.
  • the unbound iron is removed by adsorption to an alumina column or precipitated by magnesium carbonate. In either approach, centrifugation is required to separate the unbound iron from the saturated transferrin.
  • the eluant from the alumina column or the supernatant from magnesium carbonate treatment containing the transferrin-iron complex is then collected in a cup. Free iron remains absorbed on the column or in the precipitate. Then the saturated transferrin in the eluant or supernatant is assayed in an iron measuring system as discussed above for the determination of serum iron. That iron value measured represents the maximum iron the transferrin can bind.
  • serum contains chromatic components such as hemoglobin, which significantly absorb light at the measuring absorption wavelength of the iron-chromogen colored complex.
  • This problem is generally solved by forming a complex that absorbs at a wavelength at which the chromatic components of serum do not interfere.
  • Another problem as discussed above is the importance of understanding the binding of iron and transferrin at different pH values.
  • Another problem is that turbidity due to lipemia in serum interferes with spectral measurements of serum iron, as do serum proteins.
  • These problems can be solved by obtaining a serum blank measurement to correct for differences in serum turbidity and/or masking reagents such as citric acid (Garcic). Addition of DMSO is thought to lower the interference caused by lipemic samples. Denney et al.
  • US 5,763,281 states that serum or plasma can be used in the claimed method of using urea or a urea derivative and a fatty alcohol polyglycol ether.
  • Plasma is a component of blood that is generally free of blood cells, but, unlike serum, still contains the clotting proteins.
  • the laboratory needs to run a plasma instead of serum as a sample type when fast results are needed, (called "star" in clinical lab). This is because plasma can be obtained more quickly than serum.
  • a blood sample is centrifuged for 10 to 15 minutes. Then the plasma can be run in a diagnostic test.
  • serum sample after a blood sample is drawn, thirty to sixty minutes is needed to let blood clot. Then the sample must be centrifuged before it can be run.
  • the NCCLS guideline for the Determination of Serum Iron provides a procedure so that both serum and plasma can be used.
  • a precipitating-reducing reagent is made by dissolving 5 g of L-ascorbic acid in 200 mL 50% (w/v) trichloroacetic acid. Then 33 mL of 6 M hydrochloric acid is added.
  • the volume of the solution is adjusted to one liter using deionized water.
  • two milliliters of the precipitating-reducing reagent are combined and mixed with two milliliters of serum or heparinized plasma.
  • the tubes are centrifuged at about 1500g for 15 minutes. The supernatant is removed and is assayed by the NCCLS procedure. This procedure is time-consuming and difficult to automate.
  • Another disadvantage of this method is very strong acidic reagent, trichloroacetic acid was used, which is caustic and corrosive.
  • Some of the objects of the invention include providing a composition and process to eliminate the serum and plasma discrepancy that exists in the colorimetric methods of IRN and TIBC methods. Another object of the invention is to provide a composition and process for a fully automated TIBC method used for both serum and heparinized plasma as sample type. Another object of the invention is to provide an iron and TIBC assay that does not require a sample blank to subtract interference from samples.
  • the present invention is a method to determine iron and TIBC using either serum or plasma.
  • This invention is characterized by the use of a special reagent mixture as a buffer solution to eliminate the serum/plasma discrepancy, which exists in photometric IRN and TIBC method.
  • IRN iron
  • a serum or plasma sample is first combined with a buffer solution at about pH 8.5.
  • the pH is lowered to a pH which allows all the iron bound to transferrin to be released (pH about less than 5).
  • the released iron is measured by addition of a chromogen.
  • a serum or plasma sample is combined with a buffer solution at about pH 8.5 and a large excess of Fe III ions (ferric ions).
  • Fe III ions Fe III ions
  • the transferrin not already bound to iron binds to added iron. Since the Fe III is added in excess, some remains in solution.
  • an excess of chromogen that reacts with the remaining Fe III is added.
  • a blank measurement of the sample is taken on the photometer.
  • the pH is lowered to less than pH 5 to cause all of the iron bound to transferrin to be released. The released iron is measured by addition of the excess chromogen that correlates to the total iron binding capacity.
  • the invention provides fully automated methods using a novel reagent for the assay of serum iron and TIBC with both serum and plasma as appropriate sample types.
  • the present invention overcomes the discrepancy in results of serum and plasma samples. It has been found that in the IRN and TIBC methods the clotting proteins in the plasma precipitate causing turbidity and incorrect (generally high) results. The precipitation occurs upon addition of the acidic buffer to the plasma sample during the reaction(s). However, the acidic buffer is required so that bound iron may be released from transferrin. An alkaline pH buffer added prior to the addition of the acid buffer prevents the plasma protein precipitation and allows for matched serum and plasma results in both IRN and TIBC methods. The methods of the present invention use human transferrin as the calibrator material.
  • the present invention encompasses a diagnostic kit for the assay of iron comprising a first composition comprised of an alkaline buffered solution and a second composition comprised of a chromogenic indicator that reacts with iron to produce in serum or plasma a detectable change when measured in a photometer and a reducing agent to reduce Fe III to Fe II and a third composition comprising an acidic buffer.
  • the second composition may be combined with the first or third composition.
  • Any of the solutions may contain as an additive surfactants, such as Triton X, alkylammonium halides and polyhydroxyalkylene ethers to assist in the prevention of protein precipitation; reagents to minimize potential copper ion interference such as thiourea; and/or microbial inhibitors.
  • the reducing agent may be supplied as a tablet and the acid buffer added to it prior to use.
  • the first composition of the IRN kit method is a buffer having a pH of greater than 7 to a pH less than the pH that Fe (OH) does not precipitate, generally about 10.
  • the pH is at about 8-9 and most preferably the pH is about 8.6+/- 0.2.
  • This alkaline reagent preferably of TRIS buffer, could be any other buffer which will not complex with iron. Examples include Bis-Tris propane, HEPES, PIPES, Tricine Triethanolamine (TEA), GTA buffer, Diethylnolamine (DEA) and others.
  • the buffer reagent should yield a final concentration in the assay of above 0.1 M, however 0.2 M is used most often.
  • the buffer concentration in the first composition should be at about 5 - 50 times of the final concentration (e.g. .5 M - 10 M).
  • a molarity of 1 - 3 M is convenient for most dilution purposes.
  • the second composition comprises a solution of an indicator, a reducing agent to reduce Fe III to Fe II and an acetate buffer at an acidic pH.
  • the second composition may also include an agent to block copper ion interference and detergents to aid with the interference from lipemic serum samples and to aid in preventing the precipitation of proteins, particularly serum proteins.
  • the indicator may be packaged separately from both the reducing agent and the acidic buffer.
  • the acidic buffered solution may be packaged (with or without agents that block copper interference and/or detergents) as a separate composition from the composition comprising the indicator and/or reducing agent.
  • the indicator is an agent that reacts with the Fe II ion to provide a detectable signal. It is preferably a colorimetric molecule.
  • One preferred indicator is the Ferene® Indicator, (3 - 2 - pyridyl) - 5,6 - bis - 2 - (5 - furyl sulfonic acid) - 1,2,4 - triazine, disodium salt. This indicator complexes with Fe II to provide a blue complex that can be measured with a photometer.
  • Other indicators are also known from the literature and include Ferrozine®, chromazurol B, chromazurol S, Sodium bathophenanthroline (SBS), and others.
  • Ferene® or Ferrozine® are preferred due to the fast reaction kinetics and high sensitivity (High molar extinction coefficient) with these chromogens.
  • the indicator must be provided in excess of the maximum amount of iron that would be expected in any sample.
  • the preferred amount as used in the assay is greater than about 0.40 mM.
  • the indicator should have a convenient concentration for dilution purposes and for ease of manufacture, that is 10 - 1000 times the concentration required in the assay, conveniently about 4 mM to 1000 mM, preferably 200 mM to 600 mM. Most conveniently, it is about 40 mM.
  • the reducing agent must be of a type and at a concentration to reduce substantially all of the Fe III to Fe II.
  • the reducing agent is commonly ascorbic acid but other reducing agents have been used in the field and include any reducing reagent commonly used in an IRN method, such as ascorbic acid, thioglycolic acid, thiomalate, cystein, 2- mercaptoethanol, reduced glutathion, dithionite, hydroxylamine, and others.
  • the concentration used in the assay is in large excess compared to the amount of ferric ion that needs to be reduced. Generally the concentration is greater than about 15 mM. Commonly it is about 10 - 50 mM as used in the assay.
  • the reducing agent in the composition is about 10 - 100 times the concentration that is required in the assay format - that is about 100 mM - 5 M. Conveniently it may be about 300 - 500 mM.
  • the ascorbic acid may be supplied in tableted form and reconstituted with water or buffers or either composition 1 or 2 prior to use or even packaged separately.
  • the reducing agent and the indicator may be supplied with the first composition.
  • the second composition may include an acidic buffer solution.
  • the acidic buffer is packaged separately as a third composition, thereby causing less exposure to the sample.
  • the preferred buffer is an acetate buffered system.
  • the pH of this composition or the second composition, if it contains the acidic buffer should be less than 5.5, preferably about less than or equal to 4.5 so that substantially all of the iron is released from transferrin.
  • the acidic buffer concentration must be at least one time higher than the alkaline buffer concentration.
  • the acidic buffer must not complex with iron.
  • the acidic buffer must be strong enough to lower the reaction mixture pH from alkaline to acidic levels causing transferrin to release all the bound iron. However, the acid should not be too strong to precipitate the plasma proteins.
  • Many buffers can lower the pH about 4 to 5 and will not complex with iron for instance, acetate buffer, GTA buffer, oxalic acid buffer, succinate buffer, and others, preferably acetate buffer pH 3 to 5.
  • Buffers such as dicarboxylic acid, tricarboxylic acids (e.g. citric acid), ethlenediamineteraacetic acid, and phosphate that will complex with iron should be avoided.
  • the concentration of the acidic buffer as used in the assay should be about 0.2 - 2.0 M and is preferably about .4 - .8, most preferably about 0,5 M.
  • the acidic buffer as packaged in the composition can be conveniently about 1 - 10 M. If agents to reduce copper ion such as thiourea are to be included, the final concentration of such agent as used in the assay has been found effective at about 30 - 50 mM. Thus, as included in the composition, the concentration should be about 100 mM to 1 M or more. Detergents such as Triton X are effective at final assay concentrations of about 0.5 - 1.5%. Concentrations in the compositions then are conveniently about 3 - 10%. If the acidic buffer is not included in the second composition, it may be included in a separate composition.
  • the diagnostic kit comprises a first composition comprising an alkaline buffer and a second composition comprising an indicator, a reducing agent and an acidic buffer and may contain detergents and agents that minimize copper ion interference.
  • Alternative embodiments include a diagnostic kit comprising a first composition of an alkaline buffer, an indicator and a reducing agent and a second composition comprising an acid buffer and a diagnostic kit comprising a first composition comprising an alkaline buffer, a second composition comprising an indicator and reducing agent and a third composition comprising an acidic buffer. Any may include detergents, anti-microbials, preservatives or agents that minimize interference.
  • the reducing agent may be provided separately in liquid or tablet format.
  • the indicator may be provided separately.
  • Another embodiment includes the use of an indicator to Fe III. In that case a reducing agent is unnecessary, but it is important to free the Fe III from transferrin at a pH that will not cause ferric hydroxide to precipitate. Again, iron is released from transferrin at low pH values.
  • the diagnostic kit comprise a saturated solution of Fe (III), for instance a saturated solution (about greater than or equal to .35 mM) of ferric chloride to provide a Fe (III) solution of about .02mM.
  • the Fe (HI) may be supplied in a solution at pH less than about 7.
  • One preferred solution is ferric chloride in citric acid.
  • the method to assay for iron comprises adding to a sample of serum or plasma, first a solution comprising an alkaline buffer of preferentially pH 8 - 9. Next a solution (or solutions) of an indicator and a reducing agent is added. Next, an acidic solution is added.
  • the reactants may be added at different steps, except that the alkaline buffer must be added first.
  • the indicator can be included with the alkaline buffer as can the reducing agent. It is preferred to add the acidic reagent such that the sample is exposed to the acidic conditions for the shortest amount of time.
  • the method for TIBC is as follows:
  • the order of addition must be such that the alkaline buffer must be added prior to adding reagents that cause the dissociation of Fe III from transferrin.
  • the dissociation of the iron from transferrin can be caused only after the excess iron is removed or blanked.
  • the Fe III that did not bind to transferrin (since it was added in excess of the transferrin concentration) is reduced and reacted with the excess amount of indicator. The resulting color change is blanked (zeroed) on a photometer.
  • the acidic buffer is added to cause dissociation of transferrin — Fe.
  • the released iron reacts with the reducing reagent and iron indicator (which were added in excess at the step where the conditions were made alkaline) to provide and increase in absorption (increased color).
  • the alkaline and acidic buffers used in the present invention can be varied but it is critical that the combination must cause the plasma protein, particularly fibrinogen of the plasma samples to remain in solution.
  • the combinations are readily discernible to those skilled in the art.
  • Central to the invention is having a balanced buffer system in the IRN method. That is, prior to sample addition an alkaline or neutral pH buffer is added. This buffer must have the capability to stabilize plasma protein in acidic condition. Some buffers do not meet the criteria. As discussed above other buffers may be readily evaluated.
  • Read 2 minus Read 1 correspond to the serum IRN concentration.
  • Reagent 1 (Rl), the acetate buffer was added followed by 50ul of serum or plasma sample and a measurement (Read 1) was taken as a sample/reagent blank. After 3.5 minutes, reagent 2, which is iron chromogen was added. After 4 minutes, another measurement was taken (Read 2) and iron concentration correlated to read 2 minus read 1. The total reaction volume is brought to 400 ⁇ L with water. The kinetic results of a matched serum and plasma sample using the above parameters are shown in Figure 1. While the method is fast, it is evident that the plasma sample shows a discrepancy when compared to the serum sample.
  • Read 2 minus Read 1 correspond to the IRN concentration.
  • the IRN reagent composition of the present invention is shown in Table 6.
  • Example 3 The commercial IRN method was compared with the IRN method of the present invention using the parameters described in Example 1. Blood samples were freshly drawn from individuals. Serum and Lithium - heparin plasma samples were obtained from the individuals. The results of the mean of N equals five for each sample using the commercial method are presented below.
  • a method correlation of the commercial IRN method and the method of this invention was compared using 21 serum samples, calibrators and QC material.
  • the assay range is form 0 to 996 ⁇ g/dl.
  • This new iron method can be calibrated using the current commercial iron calibrator.
  • a slope of 0.9936, intercept of 3.8 ug/dl and r square of 0.9978 were obtained in the comparison.
  • the entire assay is automated on the analyzer. Instead of physically removing the excess iron from the solution, it is blanked by measuring its absorbance in the presence of a chromogen Ferene®.
  • the first step is performed using an alkaline TRIS buffer.
  • an iron saturating solution was added to saturate all the transferrin iron binding sites. In this iron-saturating reagent, the iron must be in its ferric form and must be added in excess.
  • the preferred ferric salt is ferric chloride.
  • the reaction procedure for TIBC of the present invention is:
  • the excess iron is blanked by measuring its absorbance in the presence of a chromogen.
  • the difference reading R2-R1 is used to determine TIBC, and all the interference from the sample will be cancelled out. With this new procedure no serum plasma discrepancy is observed.
  • the TIBC reagent composition of the present invention is shown in Table 9.
  • Example 5 The procedure described in Example 5 for the TIBC method of the present invention was compared using 74 matched serum samples and Li-Heparin or Na-Heparin plasma samples for the TIBC method of the present invention.
  • the range tested was 270 to 540 ⁇ g/dl. No serum/plasma discrepancy was seen.
  • Table 12 and Figures 5 and 6 show the summary of the results.
  • Example 5 The procedure described in Example 5 for the commercial TIBC and the TIBC method of the present invention were compared using 137 serum samples. The method comparison is summarized in Table 13 and Figure 7.

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Abstract

L"invention concerne un procédé et une composition servant à l"analyse (dosage) du fer et de la capacité totale de liaison du fer dans le plasma. TRIS ou une autre substance tampon est ajoutée au mélange d"analyse pour déterminer la quantité de fer et la capacité totale de liaison du fer dans le plasma.
PCT/US2001/012927 2000-04-26 2001-04-20 Reactif et procede d"analyse de «fer serique » dans le plasma WO2001081930A2 (fr)

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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006121027A1 (fr) 2005-05-12 2006-11-16 Wako Pure Chemical Industries, Ltd. Procédé de détermination de la concentration en fer
WO2009059751A1 (fr) * 2007-11-05 2009-05-14 National University Of Ireland, Maynooth Procédé de diagnostic pour déterminer une infection provoquée par ou associée à des microorganismes sécrétant des sidérophores
CN104483494A (zh) * 2014-12-22 2015-04-01 宁波美康生物科技股份有限公司 一种血清不饱和铁结合力检测试剂盒
CN109142344A (zh) * 2018-09-19 2019-01-04 南昌航空大学 一种现场快速检测分散饮用水中总铁含量的方法及装置
CN110568206A (zh) * 2019-09-12 2019-12-13 苏州普瑞斯生物科技有限公司 一种总铁结合力检测试剂盒及其制备方法
CN111007023A (zh) * 2019-12-11 2020-04-14 天津中成佳益生物科技有限公司 一种血清总铁结合力检测试剂盒及其配制方法和检测方法
CN111257549A (zh) * 2018-12-03 2020-06-09 深圳迈瑞生物医疗电子股份有限公司 检测血清中的不饱和铁结合力的试剂盒及方法
CN115290584A (zh) * 2022-08-05 2022-11-04 中拓生物有限公司 一种稳定的不饱和铁结合力测定试剂盒

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4308027A (en) * 1978-11-08 1981-12-29 R.C.C. Societa' Ricerche Di Chimica Clinica S.R.L. Method and composition for direct determination of iron in blood serum
US5219760A (en) * 1988-05-26 1993-06-15 Boehringer Mannheim Gmbh Process for the determination of iron

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4308027A (en) * 1978-11-08 1981-12-29 R.C.C. Societa' Ricerche Di Chimica Clinica S.R.L. Method and composition for direct determination of iron in blood serum
US5219760A (en) * 1988-05-26 1993-06-15 Boehringer Mannheim Gmbh Process for the determination of iron

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
CERIOTTI F ET AL: "IMPROVED DIRECT SPECIFIC DETERMINATION OF SERUM IRON AND TOTAL IRON BINDING CAPACITY" CLINICAL CHEMISTRY, vol. 26, no. 2, 1980, pages 327-331, XP002200049 ISSN: 0009-9147 *
SCHADE A.L. ET AL.: "Bound iron and unsaturated iron-binding capacity of serum; rapid and reliable quantitative determination" PROCEEDINGS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE , vol. 87, no. 1, 1954, pages 443-448, XP001073685 cited in the application *
TIETZ NORBERT W ET AL: "When is a serum iron really a serum iron? The status of serum iron measurements." CLINICAL CHEMISTRY, vol. 40, no. 4, 1994, pages 546-551, XP002198908 ISSN: 0009-9147 *
YAMANISHI HACHIRO ET AL: "Fully automated measurement of total iron-binding capacity in serum." CLINICAL CHEMISTRY, vol. 43, no. 12, December 1997 (1997-12), pages 2413-2417, XP002198906 ISSN: 0009-9147 cited in the application *

Cited By (13)

* Cited by examiner, † Cited by third party
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WO2006121027A1 (fr) 2005-05-12 2006-11-16 Wako Pure Chemical Industries, Ltd. Procédé de détermination de la concentration en fer
EP1881328A1 (fr) * 2005-05-12 2008-01-23 Wako Pure Chemical Industries, Ltd. Procédé de détermination de la concentration en fer
EP1881328A4 (fr) * 2005-05-12 2008-09-24 Wako Pure Chem Ind Ltd Procédé de détermination de la concentration en fer
US7785892B2 (en) 2005-05-12 2010-08-31 Wako Pure Chemical Industries, Ltd. Method of determining iron concentration
JP5082850B2 (ja) * 2005-05-12 2012-11-28 和光純薬工業株式会社 鉄濃度測定法
WO2009059751A1 (fr) * 2007-11-05 2009-05-14 National University Of Ireland, Maynooth Procédé de diagnostic pour déterminer une infection provoquée par ou associée à des microorganismes sécrétant des sidérophores
CN104483494A (zh) * 2014-12-22 2015-04-01 宁波美康生物科技股份有限公司 一种血清不饱和铁结合力检测试剂盒
CN109142344A (zh) * 2018-09-19 2019-01-04 南昌航空大学 一种现场快速检测分散饮用水中总铁含量的方法及装置
CN111257549A (zh) * 2018-12-03 2020-06-09 深圳迈瑞生物医疗电子股份有限公司 检测血清中的不饱和铁结合力的试剂盒及方法
CN110568206A (zh) * 2019-09-12 2019-12-13 苏州普瑞斯生物科技有限公司 一种总铁结合力检测试剂盒及其制备方法
CN111007023A (zh) * 2019-12-11 2020-04-14 天津中成佳益生物科技有限公司 一种血清总铁结合力检测试剂盒及其配制方法和检测方法
CN111007023B (zh) * 2019-12-11 2024-05-10 天津中成佳益生物科技有限公司 一种血清总铁结合力检测试剂盒及其配制方法和检测方法
CN115290584A (zh) * 2022-08-05 2022-11-04 中拓生物有限公司 一种稳定的不饱和铁结合力测定试剂盒

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