CN102866122A - Detection method of heavy metal copper in food - Google Patents
Detection method of heavy metal copper in food Download PDFInfo
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- CN102866122A CN102866122A CN2012103230985A CN201210323098A CN102866122A CN 102866122 A CN102866122 A CN 102866122A CN 2012103230985 A CN2012103230985 A CN 2012103230985A CN 201210323098 A CN201210323098 A CN 201210323098A CN 102866122 A CN102866122 A CN 102866122A
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Abstract
The invention discloses a detection method of heavy metal copper in food. The detection method comprises the following steps of: (1) selecting CCTS-g-ED as a heavy copper ion adsorbent; (2) preparing a microcolumn filled with the CCTS-g-ED; (3) preparing metal copper solutions in serial concentrations, and measuring the absorbance of the metal copper solution in each concentration by an ultraviolet and visible spectrophotometer so as to obtain a standard curve; and (4) measuring the concentration of a Cu ion in a food sample by a preconcentration-spectrophotometric method: (a) carrying out decomposition processing on the food sample, and (b) adding the obtained sample decomposition liquid into the microcolumn after adjusting the pH value of the obtained sample decomposition liquid to 5, and adding an HCl solution in a concentration of 0.9-1.1 mol/L as an eluant after finishing adsorption; and collecting all effluents as a summarized effluent, measuring the absorbance of the summarized effluent by the ultraviolet and visible spectrophotometer, substituting the absorbance into the standard curve, and thus obtaining the concentration of the Cu ion in the food sample.
Description
Technical field
The present invention relates to the detection method of a kind of heavy metal copper (Cu), relate in particular to a kind of method of utilizing preenrichment-spectrophotometric method that the Cu ion in the food is detected.
Background technology
In general, the heavy metal element in the food, a part comes from the bioconcentration of animals and plants itself; Another part then comes from the pollution that occurs in food production processing, the storage transportation.Can make after heavy metal element enters in the body that protein denaturation, enzyme lose activity, the infringement on the 26S Proteasome Structure and Function appears in histocyte, and badly influence human life security in the mode of slow poisoning.Food heavy metal pollution is one of Tough questions of facing of the whole mankind, food is carried out effective heavy metal pollution detect and seem particularly important.
Heavy metal detection method commonly used is graphite furnace atomic absorption spectrometry, ICP-AES, x ray fluorescence spectrometry etc. at present.Modern analytical technique and analytical instrument are also at development, and the detectability of element significantly reduces.But in the metal element analyse process of some low concentrations, the interference of a large amount of coexistence elements is so that direct mensuration becomes very difficult.A kind of way that addresses this problem is by development Instrument crosslinking technology, and such as ICP-MS etc., but the operation of this kind method is comparatively complicated, expense is high and time-consuming, thereby difficultly realizes conventional analysis; Another kind method then is in advance metallic ion to be carried out separation and concentration, then measures with conventional instrument.
Summary of the invention
The technical problem to be solved in the present invention provides the detection method of heavy metal copper in the food that a kind of sense cycle is short, cost is lower, utilized preenrichment-spectrophotometric method.
In order to solve the problems of the technologies described above, the invention provides the detection method of heavy metal copper in a kind of food, may further comprise the steps:
1), select de-protected modified crosslinking chitosan microball CCTS-g-ED as the heavy metal copper ion adsorbent;
2), the CCTS-g-ED that in the glass pillar, packs into, add again the pH value and be 5.0 HAc-NaAc damping fluid (0.4 mol/L) the glass pillar is filled, form microtrabeculae;
3), the metallic copper solution of preparation series concentration, and measure the absorbance of the metallic copper solution of every kind of concentration with ultraviolet-visible pectrophotometer, thereby obtain typical curve, y represents absorbance, x represents the concentration (ug/ml) of copper ion in the copper solution;
4), utilize Cu ion concentration in preenrichment-Food sample:
1., food samples is processed:
Food samples is cleared up processing, the liquid state of gained is cleared up product dilutes and constant volume, get Specimen eliminating liquid;
2., the Specimen eliminating liquid of gained is regulated pH to 5; Must regulate rear solution, solution joins above-mentioned steps 2 after will regulating) in the microtrabeculae of gained, the flow velocity of solution is that 0.5 ~ 1 mL/min(the best is 1 mL/min after the regulating and controlling); Adsorb completely, adding concentration is that 0.9 ~ 1.1 mol/L(the best is 1 mol/L) HCl solution as eluent; The flow velocity of control eluent is the best 3mL/min of being of 0.5 ~ 3 mL/min(), after wash-out is complete, collect all effluxes as the efflux after gathering, measure the absorbance of the efflux after gathering with ultraviolet-visible pectrophotometer, in the typical curve of substitution step 3) gained, final (the Cu ion concentration in the efflux after obtaining first to gather of Cu ion concentration in the food samples that obtains, again according to the volume of the efflux after gathering and the weight of food samples, through converting, finally obtain Cu ion concentration in the food samples).
Improvement as the detection method of heavy metal copper in the food of the present invention: food is animal's liver; Animal's liver is pork liver, Carp Liver or chicken gizzard.
In detection method of the present invention, step 2) the CCTS-g-ED quantity of packing in the glass pillar in must guarantee adsorption step 4) in food (animal's liver) in copper ion; Generally speaking, the concentration of copper ion is affirmed＜1000mg.kg in the animal's liver of present routine
-1
In step 2 of the present invention) in, add pH value and be 5.0 HAc-NaAc damping fluid (0.4 mol/L) CCTS-g-ED was soaked 20 ~ 28 hours, and make above-mentioned HAc-NaAc damping fluid fill pillar, the formation microtrabeculae; For subsequent use.
In step 3) of the present invention, for example can accurately be mixed with the metallic copper solution that copper ion concentration is the series concentration of 0.2 ug/ml, 0.4 ug/ml, 0.6 ug/ml, 0.8 ug/ml, and measure the absorbance of the metallic copper solution of (in 496nm place mensuration) every kind of concentration with ultraviolet-visible pectrophotometer, thereby obtain typical curve (as shown in Figure 3), y=0.9088x, y represents absorbance, and x represents the concentration (ug/ml) of copper ion in the metallic copper solution.
In step 4) of the present invention, food is cleared up processing belong to routine techniques; The liquid state of gained is cleared up product and is at first carried out the conventional acid of catching up with, and then dilutes and constant volume; Generally speaking, the liquid state of the food samples gained of 0.5g is cleared up product and is diluted constant volume (using ultrapure water) to 80 ~ 120ml after catching up with acid treatment; As Specimen eliminating liquid.Be that acetic acid-sodium acetate solution (that is, the pH value is 5.0 HAc-NaAc damping fluid) of 0.4mol/L is regulated pH to 5 with the Specimen eliminating liquid concentration of gained, must regulate afterwards solution.
In the present invention, de-protected modified crosslinking chitosan microball CCTS-g-ED is prior art, can be that 201210096788.1 " production method of crosslinked chitosan microsphere adsorbent for heavy metal " is prepared according to application number for example.
In the invention process, in order to obtain better technological parameter, the inventor had once carried out following experiment:
The selection of one, preenrichment condition:
Suitable flow velocity has important impact to the separation and concentration that Gold Samples belongs to ion.Flow velocity is too fast, and then the metallic ion in the sample can not be adsorbed fully, and the result of detection is just inaccurate; If flow velocity is too slow, then can increase the time of sample separation enrichment, reduced efficient.Therefore, select suitable flow velocity very important to the accuracy of guaranteeing experimental result.The present invention has studied different enrichments and elution flow rate, to the Cu of 5 mg/L
2+The impact of the recovery is take 1 mol/L HCl as eluant, eluent.Specific as follows:
With Cu
2+Concentration is the copper nitrate salt solusion of 5mg/L, utilize concentration for acetic acid-sodium acetate solution of 0.4mol/L (namely, the pH value is 5.0 HAc-NaAc damping fluid) regulate pH to 5, then arrive step 2 of the present invention) in the microtrabeculae of gained, the flow velocity of solution is certain value after the regulating and controlling; Adsorb complete after, add concentration and be the HCl solution of 1mol/L as eluent; The flow velocity of control eluent is certain value, and the copper ion concentration during wash-out in the efflux reduces gradually, when wash-out finishes in the efflux concentration of copper ion be zero.
The result as shown in Figure 2, CCTS-g-ED is to Cu
2+Adsorption rate reduce with the increase of enrichment flow velocity, at 0.5-1 mL/min, adsorption rate reaches more than 99.6%, so enriching rate selects 1 mL/min.Similarly, eluting rate reduces with the increase of flow velocity, and flow velocity is when 0.5-3 mL/min, and eluting rate reaches more than 99.2%, and therefore selecting elution flow rate is 3 mL/min.
Two, the impact of coexistent metallic ion
The biological sample Component comparison is complicated, comprises a lot of other micro-inorganic elements and disturbs such as Na
+, K
+, Ca
2+, Mg
2+The present invention is by adding the target method in digestion solution, namely to make its concentration be 0.5mg/L to the added metal ion.Except having studied above several metal ion species, also to Cd
2+, Pb
2+And Zn
2+To enrichment 0.5mg/L Cu
2+Impact be studied.Experimental result shows, CCTS-g-ED other ions are not inhaled or adsorbance very little.Be that these elements are to Cu in 98.6% the situation in the corresponding recovery
2+The enriching and recovering rate is impact not.
In sum, detection method of the present invention is to utilize adsorbent that testing sample (instant food sample) is carried out carrying out spectrophotometric behind the separation and concentration, have the advantage that testing cost is low, instrumentation is simple, accurate, practical, be conducive to popularizing of food heavy metal copper detection.
Description of drawings
Below in conjunction with accompanying drawing the specific embodiment of the present invention is described in further detail.
Fig. 1 is the structural drawing of part ethylenediamine (ED).
Fig. 2 is absorption and the affect figure of elution flow rate on adsorption rate and eluting rate.
Fig. 3 is the typical curve between absorbance and the copper ion concentration.
Embodiment
Be the de-protected modified crosslinking chitosan microball CCTS-g-ED of 201210096788.1 " production method of crosslinked chitosan microsphere adsorbent for heavy metal " preparation according to application number, carry out successively following steps:
A, preparation crosslinked chitosan microsphere carry out following steps successively:
1., under the room temperature, in the pulverous shitosan of 0.5g, add mass concentration and be 4% acetic acid solution 12.5ml, after shitosan fully dissolves, add again liquid paraffin 55ml, under the rotating speed of 350r/min, stirred 10 minutes;
2., step gains 1. are warming up to 55 ℃, after continuing to stir 10 minutes under the rotating speed of 350r/min, drip namely 2 of emulsifying agent span-80 0.1ml(), insulation emulsification is 10 minutes under same rotational speed; Under the effect of emulsifying agent, can form tiny shitosan drop;
3., step gains 2. are warming up to 60 ℃, adding 1.5ml formaldehyde stirring (rotating speed of 350r/min) reaction 1.5 hours; Gains are transparence;
4., to drip massfraction in step transparence gains 3. be 5% NaOH solution, when the transparence gains become white (this moment, pH value was for alkaline), stops to drip massfraction and be 5% NaOH solution; Then be warming up to 70 ℃, slowly drip epichlorokydrin 1.5ml with constant pressure funnel, stirred (rotating speed of 350r/min) insulation reaction 5 hours;
5., the product that 4. step is reacted gained filters, washing (consumption of water is 300ml), with sherwood oil suction filtration (consumption of sherwood oil is 200ml) in vacuum pump, then use the absolute ethyl alcohol suction filtration 3 times (the each consumption of absolute ethyl alcohol is 200ml), afterwards repeatedly washing is until the pH value of cleansing solution reaches neutral.
6., step gains 5. are dried to constant weight in 50 ℃ of vacuum dryers, get crosslinked chitosan microsphere (CCTS).
B, take crosslinked chitosan microsphere as parent, ethylenediamine (ED) is part, and crosslinked chitosan microsphere is carried out chemical graft, carries out successively following steps:
1., crosslinked chitosan microsphere 50mg is immersed in the hydrochloric acid solution that 125ml concentration is 1 mol/L 2 hours, after taking out in 70 ℃ of heat treated 9 hours;
2., step gains are 1. carried out first alkali cleaning, being about to step gains 1., to put into 125ml concentration be that the NaOH solution of 1oml/L soaked 2 hours; Then be washed to cleansing solution for neutral;
Carry out pickling, being about to gains after the washing, to put into 125ml concentration be that the HCl solution of 1oml/L soaked 2 hours again; Then be washed to cleansing solution for neutral;
Place 50 ℃ vacuum dryer to constant weight the gains after above-mentioned second time of the washing, obtain de-protected crosslinked chitosan microsphere (about 50mg);
3., accurately take by weighing the de-protected crosslinked chitosan microsphere of 50 mg, move in the three-necked bottle of 100 ml, add 50 ml reaction dissolvents---deionized water soaked overnight (namely soak 12 hours, thereby make de-protected crosslinked chitosan microsphere swelling);
4., ligand reagent---the ethylenediamine (ED) (its structural formula such as Fig. 1) that in step gains (going after being swelling protected crosslinked chitosan microsphere) 3., adds 250 mg, the catalyzer of 5ml---massfraction is 5% NaOH solution, under nitrogen protection in 80 ℃, heating stirring reaction 12 hours under the rotating speed of 350r/min gets the crosslinked chitosan microsphere after the modification;
5., with step 4. the crosslinked chitosan microsphere after the modification of gained with the reaction dissolvent of 20ml--deionized water soak 12 hours then use reaction dissolvent--deionized water washs, until cleansing solution is colourless, (consumption of acetone is 50ml/ time, washes 3 times respectively to wash 3 times with acetone, ether, absolute ethyl alcohol again; The consumption of ether is 50ml/ time, washes 3 times; The consumption of absolute ethyl alcohol is 50ml/ time, washes 3 times), then use distilled water (50ml/ time) washing 3 times; Be the NaOH aqueous solution soaking 2 hours of 1oml/L with 50ml concentration again, be washed to neutrality; Be that the HCl solution of 1oml/L soaked 1 hour with 50ml concentration again, be washed to neutrality; After placing 50 ℃ vacuum dryer to be dried to constant weight, obtain the about 45mg of de-protected modified crosslinking chitosan microball (being CCTS-g-ED).
After testing, CCTS-g-ED is to metallic ion Cu
2+The Dynamic Adsorption amount be 115.4mg/g.
All optionally in the following case use above-mentioned CCTS-g-ED.
The detection method of heavy metal copper in embodiment 1, the pork liver, carry out successively following steps:
1), select de-protected modified crosslinking chitosan microball CCTS-g-ED as the heavy metal copper ion adsorbent.
2), first with the glass pillar of (Φ 3 mm * 30 cm) with salpeter solution (nitric acid mixes according to the volume ratio of 1:1 with water) soaked overnight, then with ultrapure water flushing neutrality extremely, for subsequent use.Accurately take by weighing 100 mg CCTS-g-ED and pack in the above-mentioned glass pillar, fill with glass wool respectively at two ends.Add pH value and be 5.0 HAc-NaAc damping fluid (0.4 mol/L) and soaked 24 hours, make above-mentioned HAc-NaAc damping fluid fill pillar, the formation microtrabeculae; For subsequent use.
3), accurately prepare the series metal copper solution that copper ion concentration is 0.2 ug/ml, 0.4 ug/ml, 0.6 ug/ml, 0.8 ug/ml, and measure the absorbance of the metallic copper solution of (in 496nm place mensuration) every kind of concentration with ultraviolet-visible pectrophotometer, thereby obtain typical curve (as shown in Figure 3), y=0.9088x, y represents absorbance, and x represents the concentration (ug/ml) of copper ion in the metallic copper solution.
4), utilize Cu ion concentration in preenrichment-spectrophotometry sample:
1., food samples is processed:
Food to be measured is pork liver (fresh liver), and every part is the fresh liver of 0.5 g.
Cut the fresh liver of 0.5 g, put into clean polyfluortetraethylene pipe, add 5 mL HNO
3With 2 mL H
2O
2Laggard luggage tank.Program is cleared up in setting: 800 w, 15 min; 1000 w, 25 min; 0 w, 15 min.Digest complete after, after sample is cooled to room temperature (25 ℃), take out from counteract appearance, place catch up with in the electric hot plate of constant temperature (for example being 60 ℃) acid to 1 mL(namely, residue 1mL's clears up product in the polyfluortetraethylene pipe), be cooled to room temperature.
The product of clearing up of above-mentioned 1mL is put into to volumetric flask, with ultrapure water above-mentioned polyfluortetraethylene pipe is repeatedly washed (each 1.5ml, 5--10 time altogether) on a small quantity, then be settled to 100 mL with ultrapure water, as the pork liver digestion solution.
2., measure (3 repetitions are set):
Utilize concentration to regulate pH to 5 for acetic acid-sodium acetate solution (that is, the pH value is 5.0 HAc-NaAc damping fluid) of 0.4mol/L the pork liver digestion solution of gained, solution after must regulating; Then solution is added to above-mentioned steps 2 according to the enriching rate of 1 mL/min after this being regulated) in the microtrabeculae of gained; Adsorb complete after, add concentration and be the HCl solution of 1mol/L as eluent; The flow velocity of control eluent is 3 mL/min; Copper ion concentration during wash-out in the efflux reduces gradually, when wash-out finishes in the efflux concentration of copper ion be zero (use the spectrophotometric determination absorbance, concentration is learnt in conversion according to typical curve).
The remarks explanation: Fractional Collections efflux, 1mL are one section, and when the copper ion concentration in the final stage efflux was zero, enrichment finished, and the efflux that obtains before the final stage is gathered, and the efflux after must gathering (being pregnant solution) is 25ml altogether.
Adopt ultraviolet spectrophotometer to detect (detecting at the 496nm place) to above-mentioned pregnant solution, getting absorbance is 0.457; The above-mentioned y=0.9088x of substitution, thus the concentration that obtains pregnant solution is 0.503ug/ml.
Because, the pregnant solution of total 25ml, 25 * 0.503=12.575ug; Owing to be to select the fresh liver of 0.5 g as sample, therefore learn that the content of copper in the sample is 25.15 mg.kg
-1
All the other 2 parts of pork liver digestion solutions also carry out said determination, and the result of final gained is as shown in table 1.
Table 1
| The experiment sequence number | Preenrichment-spectrophotometric method (mg.kg -1?) |
| 1 | 25.15 |
| 2 | 24.82 |
| 3 | 25.09 |
Confirmatory experiment 1,
Complete fresh liver with embodiment 1 is checked that according to the ICP-AES direct measuring method result is: 24.89mg.kg
-1
The detection method of heavy metal copper in embodiment 2, the Carp Liver, carry out successively following steps:
Make the food among the embodiment 1 into Carp Liver by pork liver, all the other detection methods are equal to embodiment 1.The final detection result of 3 duplicate samples is: 15.79 mg.kg
-1, 15.40 mg.kg
-1, 15.55mg.kg
-1
Complete Carp Liver with embodiment 2 is checked that according to the ICP-AES direct measuring method result is: 15.47mg.kg
-1
The detection method of heavy metal copper in embodiment 3, the chicken gizzard, carry out successively following steps:
Make the food among the embodiment 1 into chicken gizzard by pork liver, all the other detection methods are equal to embodiment 1.The final detection result of 3 duplicate samples is: 20.50 mg.kg
-1, 20.75 mg.kg
-1, 20.30 mg.kg
-1
Confirmatory experiment 3,
Complete Carp Liver with embodiment 3 is checked that according to the ICP-AES direct measuring method result is: 20.45mg.kg
-1
Comparative Examples 1-1,
Make the enriching rate in embodiment 1 step 4) into 1.5mL/min by 1 mL/min; All the other are with embodiment 1.The pregnant solution of gained is 25ml.
Adopt ultraviolet spectrophotometer to detect (detecting at the 496nm place) to above-mentioned pregnant solution, getting absorbance is 0.366; The above-mentioned y=0.9088x of substitution, thus the concentration of the efflux after obtaining to gather is 0.403ug/ml.
Because, the pregnant solution of total 25ml, 25 * 0. 403=10.07ug; Owing to be to select the fresh liver of 0.5 g as sample, therefore learn that the content of copper in the sample is 20.14 mg.kg
-1
The accuracy of the method obviously reduces.
Comparative Examples 1-2,
Make the enriching rate in embodiment 1 step 4) into 0.5mL/min by 1 mL/min; All the other are with embodiment 1.The pregnant solution of gained is 25ml.
Adopt ultraviolet spectrophotometer to detect (detecting at the 496nm place) to above-mentioned pregnant solution, getting absorbance is 0.456; The above-mentioned y=0.9088x of substitution, thus the concentration of the efflux after obtaining to gather is 0.502ug/ml.
Because, the pregnant solution of total 25ml, 25 * 0. 502=12.55ug; Owing to be to select the fresh liver of 0.5 g as sample, therefore learn that the content of copper in the sample is 25.09 mg.kg
-1
The defective of its existence is: because the prolongation of enrichment time causes existing long defective detection time.
Comparative Examples 1-3,
Make the flow velocity of the eluent in embodiment 1 step 4) into 3.5 mL/min by 3 mL/min, all the other are with embodiment 1.The pregnant solution of gained is 25ml.
Adopt ultraviolet spectrophotometer to detect (detecting at the 496nm place) to above-mentioned pregnant solution, getting absorbance is 0.377; The above-mentioned y=0.9088x of substitution, thus the concentration of the efflux after obtaining to gather is 0.415ug/ml.
Because, the pregnant solution of total 25ml, 25 * 0. 415=10.375ug; Owing to be to select the fresh liver of 0.5 g as sample, therefore learn that the content of copper in the sample is 20.75 mg.kg
-1
The accuracy of the method obviously reduces.
Comparative Examples 1-4,
Make the flow velocity of the eluent in embodiment 1 step 4) into 2.5 mL/min by 3 mL/min, all the other are with embodiment 1.The pregnant solution of gained is 25ml.
Adopt ultraviolet spectrophotometer to detect (detecting at the 496nm place) to above-mentioned pregnant solution, getting absorbance is 0. 448; The above-mentioned y=0.9088x of substitution, thus the concentration of the efflux after obtaining to gather is 0.493ug/ml.
Because, the pregnant solution of total 25ml, 25 * 0. 493=12.325ug; Owing to be to select the fresh liver of 0.5 g as sample, therefore learn that the content of copper in the sample is 24.65 mg.kg
-1
The defective of its existence is: because the prolongation of elution time causes existing long defective detection time.
At last, it is also to be noted that what more than enumerate only is several specific embodiments of the present invention.Obviously, the invention is not restricted to above embodiment, many distortion can also be arranged.All distortion that those of ordinary skill in the art can directly derive or associate from content disclosed by the invention all should be thought protection scope of the present invention.
Claims (3)
1. the detection method of heavy metal copper in the food is characterized in that may further comprise the steps:
1), select de-protected modified crosslinking chitosan microball CCTS-g-ED as the heavy metal copper ion adsorbent;
2), the CCTS-g-ED that in the glass pillar, packs into, add again the pH value and be 5.0 HAc-NaAc damping fluid, form microtrabeculae;
3), the metallic copper solution of preparation series concentration, and measure the absorbance of the metallic copper solution of every kind of concentration with ultraviolet-visible pectrophotometer, thereby obtain typical curve, y represents absorbance, x represents the concentration ug/ml of copper ion in the copper solution;
4), utilize Cu ion concentration in preenrichment-Food sample:
1., food samples is processed:
Food samples is cleared up processing, the liquid state of gained is cleared up product dilutes and constant volume, get Specimen eliminating liquid;
2., the Specimen eliminating liquid of gained is regulated pH to 5; Must regulate rear solution, solution joins above-mentioned steps 2 after will regulating) in the microtrabeculae of gained, the flow velocity of solution is 0.5 ~ 1 mL/min after the regulating and controlling; Adsorb completely, add concentration and be the HCl solution of 0.9 ~ 1.1 mol/L as eluent; The flow velocity of control eluent is 0.5 ~ 3 mL/min, after wash-out is complete, collect all effluxes as the efflux after gathering, measure the absorbance of the efflux after gathering with ultraviolet-visible pectrophotometer, in the typical curve of substitution step 3) gained, finally obtain Cu ion concentration in the food samples.
2. the detection method of heavy metal copper in the food according to claim 1, it is characterized in that: described food is animal's liver.
3. the detection method of heavy metal copper in the food according to claim 2, it is characterized in that: described animal's liver is pork liver, Carp Liver or chicken gizzard.
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| CN104406924B (en) * | 2014-11-25 | 2017-06-13 | 苏州东辰林达检测技术有限公司 | The detection method of heavy metal in a kind of chitin |
| CN104777110B (en) * | 2015-03-18 | 2017-06-13 | 昆明理工大学 | A kind of detection method of Determination of Trace Amount Cadmium and lead |
| CN108717049A (en) * | 2018-04-17 | 2018-10-30 | 太原师范学院 | The detection method of micro copper ion concentration |
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