CN102519930A - Method for rapidly determining selenium content in plant sample - Google Patents
Method for rapidly determining selenium content in plant sample Download PDFInfo
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- CN102519930A CN102519930A CN2011104276820A CN201110427682A CN102519930A CN 102519930 A CN102519930 A CN 102519930A CN 2011104276820 A CN2011104276820 A CN 2011104276820A CN 201110427682 A CN201110427682 A CN 201110427682A CN 102519930 A CN102519930 A CN 102519930A
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Abstract
The invention discloses a method for rapidly determining the selenium content in a plant sample. According to the invention, a hydride generation-atomic fluorescence spectrometry method and a standard curve method are adopted. A mixed acid is obtained, such that an oxidation capacity is high, and over-night cold-digestion is not required. Under a certain temperature, a hot-digestion time can also be shortened, such that a detection period is greatly reduced. A national selenium standard solution is adopted, such that a standard curve can be prepared directly. The operation is simple, the accuracy us high, and a linear relationship is good. Because the atomic fluorescence spectrometry method is characterized by high interference resistance, and potassium ferricyanide has a weak effect in shielding the interferences of other elements, a detection result is almost not influenced. Therefore, the application of the potassium ferricyanide reagent can be eliminated. Therefore, reagent dosage is reduced, steps are simpler, and operation is easier.
Description
Technical field
The present invention relates to a kind of wet method and clear up the method for Se content in the fast measuring plant class sample, is to take place with hydride---AFS is measured, and belongs to the physical and chemical inspection field.
Background technology
Selenium is one of trace element of needed by human, is the important component part of biosome glutathion inside peroxidase (GSH-Px), is present in its activated centre with the form of selenocysteine.The harmful superoxide of GSH-Px in can the decomposing organism body removed free radical, prevents the impaired accumulation with heavy metal of body of cell membrane oxidation, have anti-oxidant, anticancer, anti-aging, improve effects such as body immunity, protection cardiovascular and cerebrovascular.
The whole world has more than 40 countries and regions to be in to lack selenium state, China also to have 2/3 area to lack selenium, wherein has 1/3 area to lack the selenium area for serious, as long as ground such as enshi, Ziyang, Shaanxi, Shitai County, Anhui, Fengcheng are rich selenium area.The humans and animals body lacks selenium can cause Keshan disease, Kaschin-Beck disease, and the white muscle disease of animal etc., certain cancers in addition, diseases such as region thyroid gland obstacle are also closely related with low selenium environment.China has obtained major progress through mending selenium treatment Prevention of Keshan Disease, Kaschin-Beck disease and liver cancer.
At present Determination of Selenium Contents method in the sample is had: atomic absorption spectrography (AAS), neutron activation analysis method, inductively coupled plasma mass spectrometry, ultraviolet spectrophotometry, electrochemical methods and hydride take place---atomic fluorescence spectrometry.Wherein hydride takes place---and the advantage of atomic fluorescence spectrometry is: detection limit is low, and the range of linearity is wide, and is easy and simple to handle; Operating cost ground; Hydride makes a large amount of matrix elements can not get into atomizer, thereby not only the spectrum interference is few, and chemistry disturbs also low.
Existing Se content detection method is that 200810236815.4 one Chinese patent application discloses a kind of method that adopts selenium metallic compound testing total Se content like: number of patent application; Belong to the electrochemical analysis stripping voltammetry; But it detects limit for height, and precision is low, degree of accuracy is low, and application is low.Number of patent application is that 201110020434.4 one Chinese patent application discloses a kind of total Se content detection method, is to carry out standard specimen with sodium selenite to measure, and has simplified the typical curve process for preparation.Number of patent application is the method that 200910009195.5 one Chinese patent application discloses a kind of fast measuring content of selenium in selenium polysaccharide; Belong to the optimization of colleges and universities' liquid chromatography detecting method; High performance liquid chromatography and atomic fluorescence spectrometry are relatively; Detection limit, accuracy are similar, but complicated operation, operating cost are high.
Had various kinds of document to disclose the hydride generation---Se content method in the atomic fluorescence spectrometry test sample.Like People's Republic of China's agricultural industry criteria " mensuration of full selenium in the soil " (NYT1104-2006); " mensuration of selenium in the feed " (GB/T_13883-2008); " the detection method of Se content in the tealeaves (GB/T 21729-2008); " mensuration of selenium in the food security national standard food " (GB_5009-2010) etc., the said method testing procedure is many, complicated operation, reagent dosage is big, sense cycle is long, the typical curve preparation is complicated, error is big.
So,, all exist complex steps, complicated operation, sense cycle length or cost to cross problems such as height at present to the method for the total selenium test of sample.The present invention therefore.
Summary of the invention
The method that the purpose of this invention is to provide Se content in a kind of mensuration plant class sample quick, accurate, simple to operate.
In order to solve these problems of the prior art, technical scheme provided by the invention is following:
The method of Se content in a kind of fast measuring plant class sample is characterized in that said method comprising the steps of:
(1) be the gradient selenium standard application liquid that mother liquor is mixed with 0-20 μ g/kg with selenium national standard solution; Employing hydride takes place---and atomic fluorescence spectrometry is measured selenium standard application liquid, according to the mensuration of selenium standard application liquid drawing standard curve and set up regression equation as a result;
(2) use nitric acid: the volume ratio of perchloric acid is that the acid mixture of 9-8: 1-2 clears up processing to plant class sample; The said temperature of clearing up employing is controlled at 120 ℃-180 ℃; Digestion time was controlled at 1-2 hour, carried out the acid treatment of catching up with of digestion solution then, caught up with sour temperature to select 180-210 ℃; In the digestion solution of catching up with after acid treatment is handled, add concentrated hydrochloric acid and reduce, then the reducing solution constant volume is obtained sample solution at temperature 95-110 ℃;
(3) adopting hydride to take place respectively---atomic fluorescence spectrometry is measured blank sample and sample solution; Obtain the content of selenium in the sample solution according to formula (I):
X＝(C-C
0)×V/m (I)；
In the formula: X is the content of selenium in the sample solution, and unit is every kilogram of microgram or every liter of microgram (μ g/kg or μ g/L); The concentration that C measures for the sample solution digestive juice, unit is every liter of microgram (μ g/L); C
0Measure concentration for the blank sample digestive juice, unit is every liter of microgram (μ g/L); M is sample solution quality or volume), unit is gram or milliliter (g or mL); V is a cumulative volume behind the sample solution constant volume, and unit is a milliliter (mL).
Preferably, hydride takes place in the said method---and the reductive agent that atomic fluorescence spectrometry adopts is sodium borohydride or potassium borohydride; The employing condition determination is: negative high voltage is 270V; Lamp current is 80mA; Atomization temperature is 800 ℃; The stove height is 8mm; Flow rate of carrier gas is 400ml/min; Shield gas flow speed is 800ml/min; Current-carrying is that mass concentration is 5% hydrochloric acid.
Preferably, sodium borohydride is dissolved in the sodium hydroxide solution that mass concentration is 2g/L in the said method, and constant volume shakes up and is mixed with the sodium borohydride solution that concentration is 10g/L then.
Preferably, the concentration of concentrated hydrochloric acid is 6mol/L-12mol/L in the said method step (2), and the recovery time was controlled at 10-30 minute.
Preferably, the typical curve linearly dependent coefficient requires more than 0.999 in the said method.
Preferably, the selenium concentration of the sample solution behind the constant volume is controlled in the concentration range of gradient selenium standard application liquid in the said method step (2).
Preferably, said method step (2) plant class sample carries out pulverization process in advance.
Concrete, when the present invention measures in the plant class sample Se content, carry out according to following steps:
(1) sample pre-treatments: plant class dry sample grinds with sample grinding machine or mortar after impurity elimination, clean, air-dry or oven dry, and making it all through the aperture is 0.2mm～0.6mm sieve, stores in wide-necked bottle or the valve bag, and it is subsequent use to post label.The wet appearance of gourd, fruit and vegetable class is removed the part of rotting, clean, with gauze inhale remove surface moisture after, be cut into small pieces, be milled to homogenate with sample grinding machine or mortar, store in wide-necked bottle or the valve bag, post label and deposit in the refrigerator subsequent use.
(2) reagent and preparation:
Nitric acid (top grade is pure).
Perchloric acid (top grade is pure).
Hydrochloric acid (top grade is pure).
Acid mixture preparation: nitric acid: perchloric acid=9-8: 1-2.
NaOH (top grade is pure).
Reductive agent: sodium borohydride (10g/L), with right amount of boron sodium hydride (NaBH
4), being dissolved in the sodium hydroxide solution (2g/L), constant volume shakes up then, and is subsequent use.
Current-carrying: hydrochloric acid (5%).
Single element selenium standard solution: 1000mg/L.
The configuration of selenium standard application liquid: get the selenium national standard solution mother liquor of respective volume, to become concentration be the gradient selenium standard application liquid of 0-20 μ g/kg to stepwise dilution respectively.
(3) testing sample is cleared up: accurately take by weighing and treat measuring plants class dry sample 0.2-2.0g, or the wet appearance of gourd, fruit and vegetable class 1.0-5.0g, add acid mixture 8-15ml; After mixing, place on the electric hot plate, cleared up 1-2 hour for 120 ℃-180 ℃; Catch up with nitric acid in the most solution at 180 ℃-210 ℃; To the solution white cigarette of emerging, take down cooling from electric hot plate, obtain digestion solution.In digestion solution, add 4-8ml hydrochloric acid, concentration of hydrochloric acid is 12mol/L-6mol/L, after mixing, in 95-110 ℃ of heating 10-30 minute, obtains reducing solution.With reducing solution with ultrapure water dilution constant volume in 10ml or above volumetric flask, the supernatant 5-10ml that gets the sample solution behind the constant volume is used for AFS and detects.
(4) machine is measured on drawing standard curve and the digestion solution: with AFS the selenium standard solution is measured, the drawing standard curve is also set up regression equation.Measure with the Specimen eliminating liquid of AFS after constant volume.
Acid mixture preparation in the step (2): nitric acid: perchloric acid=9-8: 1-2, ratio is high easy evaporate to dryness causes selenium to run off, and ratio is low waywardly catches up with sour terminal point.
According to total Se content in the general plant sample, be the accuracy of guaranteeing to measure in the step (2), and the typical curve good linear relationship, selenium standard application liquid maximum concentration is decided to be 20 μ g/kg.
Very competent in the step (3) because of clearing up of acid mixture, need not carry out cold clearing up to sample.
According to the character of clearing up of nitric acid-perchloric acid acid mixture, and guarantee the clear up intensity of acid mixture to plant class sample in the step (3), digestion condition is selected 120-180 ℃, in order to ensure safety, needs progressively to heat up.
In order to catch up with most nitric acid fast, catch up with sour temperature to select 180-210 ℃ in the step (3).
Adopt concentrated hydrochloric acid heating reduction hexavalent selenium to tetravalence selenium in the step (3), the time only needs 10-30 minute.
The selenium concentration of the sample solution in the step (3) behind institute's constant volume needs in the concentration range of the highest prepared selenium standard application liquid in step (2).
Gradient selenium standard application liquid can be manually prepared in the drafting of typical curve in the step (4), also can adopt AFS to prepare typical curve automatically.
The typical curve linearly dependent coefficient reaches more than 0.999 and can use in the step (4).
AFS instrument reference conditions in the step (4): negative high voltage: 270V; Lamp current: 80mA; Atomization temperature: 800 ℃; Stove is high: 8mm; Flow rate of carrier gas: 400ml/min; Shield gas flow speed: 800ml/min; Metering system: calibration curve method; Reading mode: peak area; Time delay: 1s; Reading duration: 9s.
After setting above-mentioned parameter, combustion preheater 10min～20min treats to begin to measure behind the instrument stabilizer.Elder generation's bioassay standard series, the drawing standard curve.Carry out sample again and measure, measure sample blank and sample reducing solution respectively.
Be calculated as follows the content of selenium in the sample:
X＝(C-C
0)×V/m
In the formula:
The content of selenium in the X-sample, unit are every kilogram of microgram or every liter of microgram (μ g/kg or μ g/L)
The concentration that C-sample digestive juice is measured, unit is every liter of microgram (μ g/L)
C
0-sample blank digestive juice is measured concentration, and unit is every liter of microgram (μ g/L);
M-sample mass (volume), unit is gram or milliliter (g or mL);
Cumulative volume behind the V-sample reducing solution constant volume, unit are milliliter (mL).
This method by original 2-3 days, shortens to 8-10 hour with total selenium sense cycle, detects quick, accurate, simple to operate.
Compare prior art, the present invention has following beneficial effect:
The present invention takes place through hydride---and atomic fluorescence spectrometry carries out Determination of Selenium Contents; And hydride being taken place---atomic fluorescence spectrometry is optimized; The nitration mixture oxidability of using in the method is extremely strong; Do not need cold clearing up to spend the night, and hot at a certain temperature digestion time also can reduce, shorten sense cycle greatly.Adopt directly preparation standard curve of national selenium standard solution, easy and simple to handle, accuracy is high, and linear relationship is good.Because of the strong characteristics of atomic fluorescence spectrometry antijamming capability, a little less than the effect extremely that other elements of potassium ferricyanide shielding disturb,, can reduce reagent dosage without this reagent to almost not influence of testing result, make step simpler, more easy to operate.
Embodiment:
Following examples are used to explain the present invention, but can not be used for limiting scope of the present invention.The implementation condition that adopts among the embodiment can be done further adjustment according to the condition of concrete producer, and unaccounted implementation condition is generally the condition in the normal experiment.
Embodiment 1
(1) the used AFS in this laboratory is the AFS-9230 of Beijing Jitian Instrument Co., Ltd..
(2) testing sample is cleared up: accurately take by weighing and treat measuring plants class dry sample 0.2000g, wet appearance 2.000g places the 50ml conical flask, adds acid mixture (nitric acid: 10ml perchloric acid=4: 1); Bottleneck covers the bend neck funnel, after mixing, places on the electric hot plate; Cleared up 1 hour for 180 ℃, catch up with nitric acid in the most solution, to the solution white cigarette of emerging at 210 ℃; Take down cooling from electric hot plate, obtain digestion solution.In digestion solution, add 5ml hydrochloric acid, concentration of hydrochloric acid is 6mol/L, after mixing, in 100 ℃ of heating 15 minutes, obtains reducing solution.With reducing solution with ultrapure water dilution constant volume in the 25ml volumetric flask, the supernatant 5-10ml that gets the sample solution behind the constant volume is used for AFS and detects.
(3) drawing standard curve and set up regression equation: get the selenium national standard solution mother liquor (1000mg/kg) of respective volume, stepwise dilution is to 20 μ g/kg.Automatically join the mark method with AFS; It is 0.5 μ g/kg, 2 μ g/kg, 5 μ g/kg, 10 μ g/kg, 20 μ g/kg that concentration gradient is set; To gradient selenium concentration of standard solution drawing standard curve and set up regression equation, the typical curve linearly dependent coefficient is 0.9999 with peak area.
Precision of this method and accuracy:
National standard material GBW10010 (rice), GBW10011 (wheat), GBW10012 (corn), GBW10014 (cabbage), GBW10015 (spinach), GBW10019 (apple) are carried out Determination of Selenium Contents; All in sign value scope, the result sees table 1 for 7 flat accurate values.
The mensuration result of selenium in table 1 standard substance
Can find out that by table 1 all in the calibration value scope, and precision and accuracy are very high with the measured national standard material of this method.
Embodiment 2
Relatively this method and State Standard of the People's Republic of China " mensuration of selenium in the food security national standard food " GB_5009-2010 is to result difference that same sample is surveyed.
These method detailed rules for the implementation such as embodiment 1.
The National Standard Method detailed rules for the implementation see also State Standard of the People's Republic of China's " mensuration of selenium in the food security national standard food " GB_5009-2010.
Sample detection result such as table 2 to variable concentrations
Se content relatively in table 2 National Standard Method and this method institute measuring plants class sample
Can find out that by table 2 this method for quick and national standard Law are to Se content testing result and no significant difference.
Embodiment 3
(1) experiment in used instrument with instance 1.
(2) testing sample is cleared up: accurately take by weighing and treat measuring plants class sample sample 0.5g, wet appearance 3.000g places the 50ml conical flask, adds acid mixture (nitric acid: 15ml perchloric acid=9: 1); Bottleneck covers the bend neck funnel, after mixing, places on the electric hot plate; Cleared up 1 hour for 140 ℃, catch up with nitric acid in the most solution, to the solution white cigarette of emerging at 200 ℃; Take down cooling from electric hot plate, obtain digestion solution.In digestion solution, add 4ml hydrochloric acid, concentration of hydrochloric acid is 12mol/L, after mixing, in 100 ℃ of heating 20 minutes, obtains reducing solution.With reducing solution with ultrapure water dilution constant volume in the 25ml volumetric flask, the supernatant 5-10ml that gets the sample solution behind the constant volume is used for AFS and detects.
(3) drawing standard curve and set up regression equation with embodiment 1.
Precision of this method and accuracy:
National standard material GBW10010 (rice), GBW10011 (wheat), GBW10012 (corn), GBW10014 (cabbage), GBW10015 (spinach), GBW10019 (apple) are carried out Determination of Selenium Contents; All in sign value scope, the result sees table 3 for 7 flat accurate values.
The mensuration result of selenium in table 3 standard substance
Can find out that by table 3 all in the calibration value scope, and precision and accuracy are very high with the measured national standard material of this method.
Because of digestion condition is lower than instance 1,, but the result is influenced not quite so digestion time is slightly long.
Embodiment 4
Relatively this method and State Standard of the People's Republic of China " mensuration of selenium in the food security national standard food " GB_5009-2010 is to result difference that same sample is surveyed.
These method detailed rules for the implementation such as embodiment 3.
The National Standard Method detailed rules for the implementation see also State Standard of the People's Republic of China's " mensuration of selenium in the food security national standard food " GB_5009-2010.
Sample detection result such as table 4 to variable concentrations.
Se content relatively in table 4 National Standard Method and this method institute measuring plants class sample
Can find out that by table 4 this method for quick and national standard Law are to Se content testing result and no significant difference.
Draw digestion condition when higher by above instance, digestion time can shorten greatly, but of the present invention exactly, precision is all than higher.Compare with the content method of national standard Law working sample selenium, shortened digestion time greatly, unaffected basically under the situation without screener to testing result, reduce the preparation of reagent, and operation steps.The determination experiment cycle of selenium in the plant class sample is shortened greatly, more easy to operate, measure quick, accurate.
Above-mentioned instance only is explanation technical conceive of the present invention and characteristics, and its purpose is to let the people who is familiar with this technology can understand content of the present invention and enforcement according to this, can not limit protection scope of the present invention with this.All equivalent transformations that spirit is done according to the present invention or modification all should be encompassed within protection scope of the present invention.
Claims (7)
1. the method for Se content in the fast measuring plant class sample is characterized in that said method comprising the steps of:
(1) be the gradient selenium standard application liquid that mother liquor is mixed with 0-20 μ g/kg with selenium national standard solution; Employing hydride takes place---and atomic fluorescence spectrometry is measured selenium standard application liquid, according to the mensuration of selenium standard application liquid drawing standard curve and set up regression equation as a result;
(2) use nitric acid: the volume ratio of perchloric acid is cleared up processing as the acid mixture of 9-8:1-2 to plant class sample; The said temperature of clearing up employing is controlled at 120 ℃-180 ℃; Digestion time was controlled at 1-2 hour; Carry out the acid treatment of catching up with of digestion solution then, catch up with sour temperature to select 180-210 ℃; In the digestion solution of catching up with after acid treatment is handled, add concentrated hydrochloric acid and reduce, then the reducing solution constant volume is obtained sample solution at temperature 95-110 ℃;
(3) adopting hydride to take place respectively---atomic fluorescence spectrometry is measured blank sample and sample solution; Obtain the content of selenium in the sample solution according to formula (I):
X=(C-C
0)×V/m （I）；
In the formula: X is the content of selenium in the sample solution, and unit is every kilogram of microgram or every liter of microgram (μ g/kg or μ g/L); The concentration that C measures for the sample solution digestive juice, unit is every liter of microgram (μ g/L); C
0Measure concentration for the blank sample digestive juice, unit is every liter of microgram (μ g/L); M is sample solution quality or volume), unit is gram or milliliter (g or mL); V is a cumulative volume behind the sample solution constant volume, and unit is a milliliter (mL).
2. method according to claim 1 is characterized in that hydride takes place in the said method---the reductive agent that atomic fluorescence spectrometry adopts is sodium borohydride or potassium borohydride; The employing condition determination is: negative high voltage is 270 V; Lamp current is 80 mA; Atomization temperature is 800 ℃; The stove height is 8 mm; Flow rate of carrier gas is 400 ml/min; Shield gas flow speed is 800 ml/min; Current-carrying is that mass concentration is 5% hydrochloric acid.
3. method according to claim 2 is characterized in that sodium borohydride is dissolved in the sodium hydroxide solution that mass concentration is 2 g/L in the said method, and constant volume shakes up and is mixed with the sodium borohydride solution that concentration is 10 g/L then.
4. method according to claim 1 is characterized in that the concentration of concentrated hydrochloric acid in the said method step (2) is 6mol/L-12mol/L, and the recovery time was controlled at 10-30 minute.
5. method according to claim 1 is characterized in that the requirement of typical curve linearly dependent coefficient is more than 0.999 in the said method.
6. method according to claim 1 is characterized in that the selenium concentration of the sample solution behind the constant volume in the said method step (2) is controlled in the concentration range of gradient selenium standard application liquid.
7. method according to claim 1 is characterized in that said method step (2) plant class sample carries out pulverization process in advance.
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| CN102809552A (en) * | 2012-08-15 | 2012-12-05 | 苏州硒谷科技有限公司 | Method for quickly detecting selenium of selenium-containing protein |
| CN103217407A (en) * | 2013-04-18 | 2013-07-24 | 衢州市质量技术监督检测中心 | Content measuring method for organic selenium, protein selenium, polysaccharide selenium or RNA selenium in selenium-rich rice |
| CN104122241A (en) * | 2014-07-03 | 2014-10-29 | 广西壮族自治区药用植物园 | Method for rapid determination of selenium content in tetrastigmatis hemsleyani |
| CN104655603A (en) * | 2015-02-10 | 2015-05-27 | 中国农业科学院农产品加工研究所 | Identification method for selenium-enriched rice |
| CN105021781A (en) * | 2015-08-07 | 2015-11-04 | 广东药学院 | Method for measuring selenium in food through combination of double cloud point extraction and hydride generation-atomic fluorescence spectroscopy |
| CN105372196A (en) * | 2015-12-10 | 2016-03-02 | 深圳市众凯检测技术有限公司 | Detection method of lead content of food |
| CN109269875A (en) * | 2018-11-14 | 2019-01-25 | 中国地质科学院地球物理地球化学勘查研究所 | A method of inhibiting selenium volatilization in plant sample digestion process |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102809552A (en) * | 2012-08-15 | 2012-12-05 | 苏州硒谷科技有限公司 | Method for quickly detecting selenium of selenium-containing protein |
| CN103217407A (en) * | 2013-04-18 | 2013-07-24 | 衢州市质量技术监督检测中心 | Content measuring method for organic selenium, protein selenium, polysaccharide selenium or RNA selenium in selenium-rich rice |
| CN104122241A (en) * | 2014-07-03 | 2014-10-29 | 广西壮族自治区药用植物园 | Method for rapid determination of selenium content in tetrastigmatis hemsleyani |
| CN104655603A (en) * | 2015-02-10 | 2015-05-27 | 中国农业科学院农产品加工研究所 | Identification method for selenium-enriched rice |
| CN105021781A (en) * | 2015-08-07 | 2015-11-04 | 广东药学院 | Method for measuring selenium in food through combination of double cloud point extraction and hydride generation-atomic fluorescence spectroscopy |
| CN105372196A (en) * | 2015-12-10 | 2016-03-02 | 深圳市众凯检测技术有限公司 | Detection method of lead content of food |
| CN105372196B (en) * | 2015-12-10 | 2018-05-08 | 深圳市众凯检测技术有限公司 | A kind of detection method of Lead Content in Foodstuff |
| CN109269875A (en) * | 2018-11-14 | 2019-01-25 | 中国地质科学院地球物理地球化学勘查研究所 | A method of inhibiting selenium volatilization in plant sample digestion process |
| CN109269875B (en) * | 2018-11-14 | 2021-05-18 | 中国地质科学院地球物理地球化学勘查研究所 | Method for inhibiting selenium volatilization in plant sample digestion process |
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