CN102590195A - Method for testing nitrite and nitrate in pickled vegetables - Google Patents
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Abstract
The invention belongs to the field of chemistry, and relates to a method for testing the nitrite and nitrate in pickled vegetables. The method for rapidly testing the nitrite and nitrate in pickled vegetables comprises the following steps that: (1) the nitrite and the nitrate in pickled vegetable are extracted by ultrasonic waves; (2) a calibration curve is drawn by using nitrosonitrogen (NO2-) standard solution; and a calibration curve is drawn by using nitronitrogen (NO3-) standard solution; (3) by diazotizing sulfamide, the nitrite in the pickled vegetable is coupled with hydrochloric acid N-(1-naphthyl)ethylenediamine, so that rosy azo dye is produced, and the content of the nitrite is determined at 540nm; (4) the nitrate in the pickled vegetable is reduced to nitrite by a cadmium column, the nitrite is determined, the determined nitrite in the same sample is subtracted, and the nitrate content is worked out.
Description
Technical field
The invention belongs to chemical field, be applicable to the mensuration of nitrite, nitrate in the sour salted vegetables.
Background technology
Nitrite, English name Nitrite, abbreviation NIT is commonly called as salt made from earth containing a comparatively high percentage of sodium chloride, is a kind of White-opalescent crystal, and shape is similar with salt.Nitrite is a carcinogen, and severe toxicity is arranged, and toxicity symptom is enterogenous cyanosis disease, cyanosis disease, black mouth disease, and the nitrite of eating 0.3～0.5 gram can cause poisons even death.The nitrite poisoning morbidity rapidly; In general 1～3 hour latent period, the principal feature of poisoning is because histanoxia causes the cyanosis phenomenon, and is livid purple like lip, the tip of the tongue, finger tip; Weight person's eye conjunctiva, face and whole skin are livid purple; Occur dizziness, headache, weak, palpitate quickly, symptoms such as drowsiness or irritated, expiratory dyspnea, nauseating, vomiting, stomachache, diarrhoea, stupor, convulsions, gatism appear in severe patient, until death.Pickle, stored easy generation nitrite such as ripe leftovers of a specified duration, and contained multiple nitrate in the vegetables, particularly the big vegetables of nitrogen fertilizer application amount contain more nitrate.The enzyme of vegetables self can reduce nitrate, changes into nitrite; Vegetables of cooking or leftovers were placed of a specified duration, under the effect of bacterium, also can produce nitrite.
Reason owing to raw material and processing technology; All contain a certain amount of nitrite and nitrate in any curing food inevitably; The acid salted vegetables is a kind of plant curing food; Also contain nitrite, nitrate, how to monitor, control nitrite, nitrate in the sour salted vegetables, average family, citizen are difficult to accomplish.The present invention is through detecting the nitrite in the sour salted vegetables curing process, the content of nitrate; Seek nitrite, nitrate Changing Pattern in the sour salted vegetables curing process; Confirm that sour salted vegetables best safety eats the phase, reduce the injury of nitrite in the sour salted vegetables, nitrate human body.Mainly there is sodium nitrite molecular formula NaNO in nitrite in the acid salted vegetables with the sodium nitrite form
2, sodium nitrite is extremely strong carcinogen; Nitrate mainly exists with the sodium nitrate form, molecular formula NaNO
3, the nitrate in the curing food very easily is converted into nitrite, thereby human body is produced harm.
The acid of acid salted vegetables mouthfeel is refreshing, and taste is unique, is the batching commonly used of house travelling, local characteristic snack, also is the necessity of popular life.Acid salted vegetables manufacture craft is simple, and is salted with green vegetables (Yunnan and other places claim bitter dish), and fresh vegetable is through sun solarization after one, two day; Clean, dry, optionally prescind or chop up, add spices such as salt, chilli powder, anise seed, Cao Jiao, anise; Mix thoroughly, spray a little edible wine, move in the cylinder; Compress, seal, it is edible to open jar after 25 days.
The invention provides a kind of method of monitoring nitrite, nitrate in the sour salted vegetables, can fast, accurately detect the content of nitrite, nitrate in the sour salted vegetables.The present invention uses full-automatic flow injection analyzer; Measure nitrite, nitrate in the sour salted vegetables; Technological precedence, finding speed is fast, the result is accurate, is particularly useful for the detection of carcinogen sodium nitrite in the sour salted vegetables, sodium nitrate content, belongs to the scientific and technical innovation project.At present, do not find the report of nitrite, nitrate method in the related detection acid salted vegetables both at home and abroad as yet.
The present invention is intended to overcome the defective of prior art, provides a kind of employing full-automatic flow injection analyzer, measures the method for nitrite, nitrate in the sour salted vegetables, detection limit sodium nitrite 0.14mg/kg, sodium nitrate 0.17mg/kg.National standard " mensuration of nitrite and nitrate in the food safety and sanitation standard food " GB5009.33-2010 has stipulated the assay method of nitrite and nitrate, the mensuration (three therapeutic methods of traditional Chinese medicine) of nitrite and nitrate in the chromatography of ions (first method), AAS (second method), the milk and milk products; National standard " catsup and pickled vegetables hygienic standard " GB2714-2003, " analytical approach of catsup and pickled vegetables hygienic standard " GB/T5009.54-2003 have stipulated limiting the quantity of and assay method of nitrite in the catsup and pickled vegetables, and content of nitrite is (with NaNO
2Meter)≤and 20mg/kg, assay method is the method for GB5009.33 regulation.Existing nitrite, the standard of nitrate, the method for detecting, all different with used instrument and the determination step of the present invention, the present invention has the novelty and the practicality of distinctness.
Summary of the invention
Nitrite, nitrate method in the sour salted vegetables of mensuration according to the invention, form by following steps:
One, quantitatively takes by weighing sour salted vegetables sample, place that homogenizer is smashed to pieces, mixing, with nitrite, the nitrate in the ultrasonic extraction acid salted vegetables.
Two, draw the nitrite calibration curve: use nitroso-nitrogen (NO
2 -) standard solution, being mixed with concentration is the standard operation solution of 0.0,2.0,5.0,10.0,20.0,50.0,100.0 μ g/L, is horizontal ordinate with the nitroso-nitrogen concentration, nitroso-nitrogen peak area is an ordinate, the computer automatic drafting calibration curve.
Use nitrate (NO
3 -) standard solution, being mixed with concentration is the standard operation solution of 0.0,2.0,20.0,100.0,200.0,500.0,1000.0 μ g/L, is horizontal ordinate with nitrate concentration, the nitrate peak area is an ordinate, the computer automatic drafting calibration curve.
Three, quantitatively add sulfamide diazotising according to nitrite in the sour salted vegetables of per 1.0 grams,, generate the rose azo dyes, measure content of nitrite at 540nm with hydrochloric acid N-(1-naphthyl) ethylenediamine coupling.
Content of nitrite calculates by (1) formula in the acid salted vegetables sample:
In the formula:
X
1---content of nitrite is (with NaNO in the sour salted vegetables
2Meter), unit is every kilogram (mg/kg) of milligram;
C
X1---sample solution Central Asia nitrate concentration, unit is every liter of microgram (μ g/L);
C
01---blank test solution Central Asia nitrate concentration, unit is every liter of microgram (μ g/L);
V---sample solution is measured volume, and unit is a milliliter (mL);
V
1---sample solution is obtained volume, and unit is a milliliter (mL);
V
0---sample solution constant volume, unit are milliliter (mL);
M---sample mass, unit is gram (g);
k
1---nitroso-nitrogen is scaled the coefficient of sodium nitrite, 4.929.
The result keeps 2 position effective digitals behind the radix point.
Four, be nitrite according to the nitrate in the sour salted vegetables sample of per 1.0 grams through Cadmium column reduction; Quantitatively add sulfamide diazotising; With hydrochloric acid N-(1-naphthyl) ethylenediamine coupling, generate the rose azo dyes, measure nitrite at 540nm; Deduct the content of nitrite of having measured in the same sample, convert nitrate content.
Nitrate content calculates by (2) formula in the acid salted vegetables sample:
In the formula:
X
2---nitrate content is (with NaNO in the sour salted vegetables
3Meter), unit is every kilogram (mg/kg) of milligram;
X
1---content of nitrite is (with NaNO in the sour salted vegetables
3Meter), unit is every kilogram (mg/kg) of milligram;
C
X2---sample solution Central Asia nitrate concentration, unit is every liter of microgram (μ g/L);
C
02---blank test solution Central Asia nitrate concentration, unit is every liter of microgram (μ g/L);
V---sample solution is measured volume, and unit is a milliliter (mL);
V
2---sample solution is obtained volume, and unit is a milliliter (mL);
V
0---sample solution constant volume, unit are milliliter (mL);
M---sample mass, unit is gram (g);
k
1---nitroso-nitrogen is scaled the coefficient of sodium nitrite, 4.929;
k
2---sodium nitrite is scaled the coefficient of sodium nitrate, 1.232.
The result keeps 2 position effective digitals behind the radix point.
The present invention has overcome the defective of prior art; Use full-automatic flow injection analyzer; Measure carcinogen nitrite, nitrate in the sour salted vegetables, technological precedence, finding speed is fast, the result is accurate, is applicable to the detection of food security project; Filled up the blank of sour salted vegetables check, significant to protection consumer health and food security work.
Principal feature of the present invention:
1, use full-automatic flow injection analyzer to measure nitrite, nitrate content in the sour salted vegetables, the sample single treatment is measured secondary and is accomplished.
2, the measurement detection limit is low, and measure nitrite in the sour salted vegetables, nitrate content detection limit: nitrite is (with NaNO
2Meter) 0.14mg/kg, nitrate are (with NaNO
3Meter) 0.17mg/kg.
3, accuracy of measurement meets the demands, and measures nitrite in the sour salted vegetables, nitrate content recovery of standard addition: nitroso-nitrogen recovery of standard addition 93.1～109.2%, and nitrate recovery of standard addition 82.1～118.8%, it is accurate to measure the result.
4, measuring repeatability meets the requirements; Measure sodium nitrite measurement relative standard deviation (RSD) 0.74～3.34% in the sour salted vegetables, sodium nitrate is measured relative standard deviation (RSD) 0.59～3.90%, measures relative standard deviation (RSD)≤4.00%; Error at measurment is little, and the result is accurate.
Embodiment
Embodiment 1:
1 measuring principle
1.1 nitrite measuring principle
Nitrite in the acid salted vegetables sample with hydrochloric acid N-(1-naphthyl) ethylenediamine coupling, generates the rose azo dyes through to sulfamide diazotising, measures content of nitrite at 540nm.
1.2 nitrate measuring principle
Nitrate in the acid salted vegetables sample is nitrite through Cadmium column reduction; Warp is to sulfamide diazotising; With hydrochloric acid N-(1-naphthyl) ethylenediamine coupling, generate the rose azo dyes, measure nitrite at 540nm; Deduct the content of nitrite of having measured in the same sample, convert nitrate content.
2 use instrument and reagent
2.1 instrument
Full-automatic flow injection analyzer, model: FIA-6000
+, Beijing Jitian Instrument Co., Ltd. produces, and the instrument service condition is seen table 1.
The full-automatic flow injection analyzer condition of work of table 1
| Project | Condition | Project | Condition |
| Sample introduction needle scavenging period (S) | 10 | Appearance time (S) | 20 |
| Time of arrival (S) | 25 | Peak width (S) | 60 |
| Inject time (S) | 30 | The current-carrying position | Z2 |
| Sample injection time (S) | 21 | The cleaning position | Z3 |
| Scavenging period (S) | 20 | Wash the pin position | Z2 |
| The sample cycle time (S) | 60 | Pump speed | 35 |
The numerical control supersonic washer, model: KH-300DE, Kunshan standing grain wound ultrasonic instrument company limited produces, and the instrument service condition is seen table 2.
Table 2 ultrasonic cleaner condition of work
| Time (min) | Temperature (℃) | Power (%) |
| 20 | 20 | 50 |
2.2 use reagent
2.2.1 experimental water is a deionized water, chemical reagent is that top grade is pure;
2.2.2 hydrochloric acid (HCl:35-38%);
2.2.3 ammoniacal liquor (NH
3: 25-28%);
2.2.4 phosphoric acid (H
3PO
4:>=85%);
2.2.5 NaOH;
2.2.6 two ethylenediamine hydrate tetraacethyl disodium (Na
2EDTA2H
2O);
2.2.7 sulfanilamide (SN) (P-aminobenzene-sulfonamide, C
6H
8N
2O
2S
2);
(2.2.8N-1-naphthyl) ethylenediamine-hydrochloride (C
12H
16Cl
2N
2);
2.2.9 nitroso-nitrogen standard solution: national standard solution, 1000.0mg/L;
2.2.10 nitrate standard solution: national standard solution, 100.0mg/L;
2.2.11 ammonium chloride buffer (pH8.5): in the 500mL water, add 105mL hydrochloric acid
(2.2.2), 95mL ammoniacal liquor (2.2.3), 1.0g two ethylenediamine hydrate tetraacethyl disodiums
(2.2.6), mixing, transferring to pH with 8.0g NaOH (2.2.5) is 8.5, is settled to 1 liter.
2.2.12 sulfanilamide (SN) developer: add 100mL phosphoric acid (2.2.4), 40.0g sulfanilamide (SN) (2.2.7), 1.0g N-(1-naphthyl) ethylenediamine-hydrochloride (2.2.8) in the 600mL water, be stirred to whole dissolvings, be settled to 1L, be stored in the brown bottle.
3 measure
3.1 sour salted vegetables test liquid preparation
Weighing acid salted vegetables test portion 10.0g (being accurate to 0.0001g) smashs 2min to pieces in homogenizer, move in the 100mL glass volumetric flask, and water is settled to scale, mixing, and ultrasonic extraction 20min leaves standstill.
Obtain the 5mL supernatant to the 50mL volumetric flask, water is settled to scale, and mixing is measured nitrite.When content of nitrite is higher, answer dilution metering.
Obtain the 1mL supernatant to the 50mL volumetric flask, water is settled to scale, and mixing is measured nitrate.When nitrate content is higher, answer dilution metering.
3.2 nitroso-nitrogen (NO
2 -) the calibration curve drafting
Use nitroso-nitrogen (NO
2 -) standard solution (2.2.9), being mixed with concentration is the standard operation solution of 0.0,2.0,5.0,10.0,20.0,50.0,100.0 μ g/L, is used for the mensuration of nitrite.Under table 1 instrument condition of work; Connecting deionized water (current-carrying), damping fluid (2.2.11), developer (2.2.12), measure nitroso-nitrogen absorption peak, is horizontal ordinate with the nitroso-nitrogen concentration; Nitroso-nitrogen peak area is an ordinate, the computer automatic drafting calibration curve.3.3 nitrate (NO
3 -) the calibration curve drafting
Use nitrate (NO
3 -) standard solution (2.2.10), being mixed with concentration is the standard operation solution of 0.0,2.0,20.0,100.0,200.0,500.0,1000.0 μ g/L, is used for the mensuration of nitrate.Measuring condition is measured the nitrate absorption peak with 3.2, is horizontal ordinate with nitrate concentration, and the nitrate peak area is an ordinate, the computer automatic drafting calibration curve.
3.4 nitrite is measured
Under 3.2 conditions, measure the sour salted vegetables sample solution of 3.1 preparations, content of nitrite calculates by (3) formula in the sour salted vegetables sample:
In the formula:
X
1---content of nitrite is (with NaNO in the sour salted vegetables
2Meter), unit is every kilogram (mg/kg) of milligram;
C
X1---sample solution Central Asia nitrate concentration, unit is every liter of microgram (μ g/L);
C
01---blank test solution Central Asia nitrate concentration, unit is every liter of microgram (μ g/L);
V---sample solution is measured volume, and unit is a milliliter (mL);
V
1---sample solution is obtained volume, and unit is a milliliter (mL);
V
0---sample solution constant volume, unit are milliliter (mL);
M---sample mass, unit is gram (g);
k
1---nitroso-nitrogen is scaled the coefficient of sodium nitrite, 4.929.
The result keeps 2 position effective digitals behind the radix point.
3.5 nitrate is measured
Under 3.3 conditions, measure the sour salted vegetables sample solution of 3.1 preparations, nitrate content calculates by (4) formula in the sour salted vegetables sample:
In the formula:
X
2---nitrate content is (with NaNO in the sour salted vegetables
3Meter), unit is every kilogram (mg/kg) of milligram;
X
1---content of nitrite is (with NaNO in the sour salted vegetables
3Meter), unit is every kilogram (mg/kg) of milligram;
C
X2---sample solution Central Asia nitrate concentration, unit is every liter of microgram (μ g/L);
C
02---blank test solution Central Asia nitrate concentration, unit is every liter of microgram (μ g/L);
V---sample solution is measured volume, and unit is a milliliter (mL);
V
2---sample solution is obtained volume, and unit is a milliliter (mL);
V
0---sample solution constant volume, unit are milliliter (mL);
M---sample mass, unit is gram (g);
k
1---nitroso-nitrogen is scaled the coefficient of sodium nitrite, 4.929;
k
2---sodium nitrite is scaled the coefficient of sodium nitrate, 1.232.
The result keeps 2 position effective digitals behind the radix point.
4 measure interpretation of result
4.1 natrium nitrosum, sodium nitrate measurement result see Table 3 in the sour salted vegetables sample.
Natrium nitrosum, sodium nitrate measurement result in the sour salted vegetables sample of table 3
| Sample | Sodium nitrite, mg/kg | Sodium nitrate, % |
| The 2010-10-sample | 8.06 | 0.84 |
| The 2010-11-sample | 17.80 | 0.75 |
4.2 sodium nitrite, sodium nitrate changes of contents rule in the sour salted vegetables sample
To carry out sodium nitrite, sodium nitrate detection with a collection of sour salted vegetables; Detect once every day; The result proves: under the normal temperature (20 ℃); Acid salted vegetables content of nitrite in curing process increases gradually, and content of nitrite was the highest in 9～13 days, surpass national standard " catsup and pickled vegetables hygienic standard " GB2714-2003 regulation " content of nitrite is (with NaNO
2Meter)≤20mg/kg " limit standard, what have surpasses 2～3 times of limit standards.This moment, the food acids salted vegetables must be harmful to health; After normally pickling 15 days, content of nitrite reduces gradually in the sour salted vegetables, and is safe to eat after 25 days.Content of sodium nitrite sees Table 4 with the variation of salting period in the acid salted vegetables.
The variation of sodium nitrite, sodium nitrate content in the sour salted vegetables curing process of table 4
The embodiment technical identification:
1 detection limit confirmatory experiment
1.1 blank solution detection limit experiment
Measure blank solution 10 times, basis of calculation deviation sees Table 5.
Table 5 blank solution detection limit experimental result (n=10, μ g/L)
1.2 get sodium nitrite, sodium nitrate calibration curve minimum point 2.0 μ g/L mensuration 10 times, basis of calculation deviation is seen table 6.
Table 6 minimum point concentration detection limit experimental result (n=10, μ g/L)
3 times with minimum point concentration determination value standard deviation serves as to measure detection limit, nitrite detection limit 0.56 μ g/L, and nitrate detection limit 0.33 μ g/L a little more than the instrument acceptance index, belongs to normal range.Be converted into that the sodium nitrite detection limit is 0.14mg/kg in the sour salted vegetables sample, sodium nitrate detects and is limited to 0.17mg/kg.
The experiment of 2 accuracy of measurement
Before the sour salted vegetables sample of extraction, quantitatively add nitroso-nitrogen, nitrate standard solution, carry out recovery experiment, measure the recovery and see table 7, table 8.
Table 7 nitroso-nitrogen recovery of standard addition (n=20, μ g/L)
Table 8 nitrate recovery of standard addition (n=20, μ g/L)
Experimental result: measure nitroso-nitrogen recovery of standard addition 93.1～109.2%, measure nitrate recovery of standard addition 82.1～118.8%.
3 measure repeated experiment
Select for use different sour salted vegetables samples to carry out measurement reproducibility experiment, measure 21 times, calculate each element relative standard deviation (RSD), the results are shown in Table 9, table 10.
Table 9 nitrite measurement stability experiment (mode standard, n=21, μ g/L)
Table 10 nitrate measurement stability experiment (mode standard, n=21, μ g/L)
Sodium nitrite is measured relative standard deviation (RSD) 0.74～3.34%, and sodium nitrate is measured relative standard deviation (RSD) 0.59～3.90%, measures relative standard deviation (RSD)≤4.00%, measures good reproducibility, and the result is accurate.
Claims (6)
1. method that detects nitrite, nitrate in the sour salted vegetables is characterized in that being made up of following steps:
One, quantitatively takes by weighing sour salted vegetables sample, place that homogenizer is smashed to pieces, mixing, with nitrite, the nitrate in the ultrasonic extraction acid salted vegetables.
Two, use nitroso-nitrogen (NO
2 -) standard solution, being mixed with concentration is the standard operation solution of 0.0,2.0,5.0,10.0,20.0,50.0,100.0 μ g/L, full-automatic flow injection analyzer is drawn calibration curve.Use nitrate (NO
3 -) standard solution, being mixed with concentration is the standard operation solution of 0.0,2.0,20.0,100.0,200.0,500.0,1000.0 μ g/L, draws calibration curve.
Three, sour salted vegetables sample Central Asia nitrate with hydrochloric acid N-(1-naphthyl) ethylenediamine coupling, generates the rose azo dyes through to sulfamide diazotising, measures content of nitrite at 540nm.
Four, nitrate was nitrite through Cadmium column reduction during acid salted down, through to sulfamide diazotising, with hydrochloric acid N-(1-naphthyl) ethylenediamine coupling, generated the rose azo dyes and measured nitrite, deducted nitrite in the same sample, obtained nitrate content.
2. a kind of according to claim 1 method that detects nitrite, nitrate in the sour salted vegetables is characterized in that using full-automatic flow injection analyzer to measure carcinogen nitrite, nitrate content in the sour salted vegetables.
3. a kind of according to claim 1 method that detects nitrite, nitrate in the sour salted vegetables; It is characterized in that finding rule: under the normal temperature (20 ℃) through test experience; Acid salted vegetables content of nitrite in curing process increases gradually, and content of nitrite was the highest in 9～13 days, pickle 15 days after; Content of nitrite reduces gradually in the acid salted vegetables, and is safe to eat after 25 days.
4. a kind of according to claim 1 method that detects nitrite, nitrate in the sour salted vegetables, it is characterized in that detection limit is low: the nitrite detection limit is (with NaNO
2Meter) 0.14mg/kg, nitrate detection limit are (with NaNO
3Meter) 0.17mg/kg.
5. a kind of according to claim 1 method that detects nitrite, nitrate in the sour salted vegetables is characterized in that nitroso-nitrogen recovery of standard addition 93.1～109.2%, measures nitrate recovery of standard addition 82.1～118.8%, and it is accurate to measure the result.
6. a kind of according to claim 1 method that detects nitrite, nitrate in the sour salted vegetables; It is characterized in that measuring repeatability meets the requirements; Measure sodium nitrite measurement relative standard deviation (RSD) 0.74～3.34% in the sour salted vegetables, sodium nitrate is measured relative standard deviation (RSD) 0.59～3.90%, measures relative standard deviation (RSD)≤4.00%; Error at measurment is little, and the result is accurate.
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| CN109557267A (en) * | 2018-12-10 | 2019-04-02 | 荀佳钰 | A kind of nitrite concentration research method that pickling of food process generates |
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