CN103884796B - Method for detecting total propionic acid content of hybrid feed additive - Google Patents
Method for detecting total propionic acid content of hybrid feed additive Download PDFInfo
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- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 title claims abstract description 386
- 235000019260 propionic acid Nutrition 0.000 title claims abstract description 192
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 title claims abstract description 192
- 238000000034 method Methods 0.000 title claims abstract description 25
- 239000003674 animal food additive Substances 0.000 title abstract description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 47
- XBDQKXXYIPTUBI-UHFFFAOYSA-M Propionate Chemical compound CCC([O-])=O XBDQKXXYIPTUBI-UHFFFAOYSA-M 0.000 claims abstract description 47
- 239000000706 filtrate Substances 0.000 claims abstract description 24
- 238000001514 detection method Methods 0.000 claims abstract description 19
- 238000012937 correction Methods 0.000 claims abstract description 13
- 239000012528 membrane Substances 0.000 claims abstract description 10
- 150000001875 compounds Chemical class 0.000 claims description 36
- 239000000243 solution Substances 0.000 claims description 32
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 26
- 239000002671 adjuvant Substances 0.000 claims description 17
- 239000003795 chemical substances by application Substances 0.000 claims description 17
- LDHQCZJRKDOVOX-NSCUHMNNSA-N crotonic acid Chemical compound C\C=C\C(O)=O LDHQCZJRKDOVOX-NSCUHMNNSA-N 0.000 claims description 16
- LDHQCZJRKDOVOX-UHFFFAOYSA-N trans-crotonic acid Natural products CC=CC(O)=O LDHQCZJRKDOVOX-UHFFFAOYSA-N 0.000 claims description 16
- 239000006228 supernatant Substances 0.000 claims description 15
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 13
- 235000019253 formic acid Nutrition 0.000 claims description 13
- 235000019441 ethanol Nutrition 0.000 claims description 12
- 239000000284 extract Substances 0.000 claims description 12
- 239000012086 standard solution Substances 0.000 claims description 12
- 239000012224 working solution Substances 0.000 claims description 11
- XJMWHXZUIGHOBA-UHFFFAOYSA-N azane;propanoic acid Chemical compound N.CCC(O)=O XJMWHXZUIGHOBA-UHFFFAOYSA-N 0.000 claims description 9
- 238000003965 capillary gas chromatography Methods 0.000 claims description 9
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 8
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 8
- 125000005909 ethyl alcohol group Chemical group 0.000 claims description 8
- 238000002347 injection Methods 0.000 claims description 5
- 239000007924 injection Substances 0.000 claims description 5
- 229910021529 ammonia Inorganic materials 0.000 claims description 4
- 239000000377 silicon dioxide Substances 0.000 claims description 4
- 235000012239 silicon dioxide Nutrition 0.000 claims description 4
- 229910052902 vermiculite Inorganic materials 0.000 claims description 4
- 239000010455 vermiculite Substances 0.000 claims description 4
- 235000019354 vermiculite Nutrition 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 claims description 3
- 230000020477 pH reduction Effects 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 4
- 238000004587 chromatography analysis Methods 0.000 abstract description 3
- 239000011259 mixed solution Substances 0.000 abstract description 3
- 231100000252 nontoxic Toxicity 0.000 abstract description 3
- 230000003000 nontoxic effect Effects 0.000 abstract description 3
- 238000001914 filtration Methods 0.000 abstract 2
- 239000000126 substance Substances 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 48
- 238000012360 testing method Methods 0.000 description 10
- 230000003449 preventive effect Effects 0.000 description 8
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- 239000007789 gas Substances 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 5
- 239000000203 mixture Substances 0.000 description 4
- 238000002203 pretreatment Methods 0.000 description 4
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- BCZXFFBUYPCTSJ-UHFFFAOYSA-L Calcium propionate Chemical compound [Ca+2].CCC([O-])=O.CCC([O-])=O BCZXFFBUYPCTSJ-UHFFFAOYSA-L 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 235000010331 calcium propionate Nutrition 0.000 description 3
- 239000004330 calcium propionate Substances 0.000 description 3
- 238000010812 external standard method Methods 0.000 description 3
- 238000010813 internal standard method Methods 0.000 description 3
- BWILYWWHXDGKQA-UHFFFAOYSA-M potassium propanoate Chemical compound [K+].CCC([O-])=O BWILYWWHXDGKQA-UHFFFAOYSA-M 0.000 description 3
- 235000010332 potassium propionate Nutrition 0.000 description 3
- 239000004331 potassium propionate Substances 0.000 description 3
- 239000012488 sample solution Substances 0.000 description 3
- JXKPEJDQGNYQSM-UHFFFAOYSA-M sodium propionate Chemical compound [Na+].CCC([O-])=O JXKPEJDQGNYQSM-UHFFFAOYSA-M 0.000 description 3
- 235000010334 sodium propionate Nutrition 0.000 description 3
- 239000004324 sodium propionate Substances 0.000 description 3
- 229960003212 sodium propionate Drugs 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 238000004817 gas chromatography Methods 0.000 description 2
- 229910052500 inorganic mineral Inorganic materials 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000011707 mineral Substances 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 238000003822 preparative gas chromatography Methods 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000012159 carrier gas Substances 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 238000009434 installation Methods 0.000 description 1
- 239000013067 intermediate product Substances 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
The invention discloses a method for detecting total propionic acid content of a hybrid feed additive. The method comprises the steps of 1) making a corresponding relation standard curve of the concentration and a peak area of propionic acid measured by a gas chromatographic method, or obtaining a correction factor of propionic acid relative to an internal standard substance; 2) converting propionate in a sample to be detected into free propionic acid, and extracting the propionic acid through absolute ethyl alcohol to obtain a mixed solution; 3) filtering supernate of the mixed solution obtained in the step 2) through a disposable organic filtering membrane, and measuring the peak area of the propionic acid in filtrate by the gas chromatographic method; 4) calculating the concentration of the propionic acid in the filtrate according to the corresponding relation standard curve of the concentration and the peak area of the propionic acid as well as the peak area of the propionic acid in the filtrate obtained in the step 3), and then calculating the total propionic acid content of the sample to be detected. According to the method, the pretreatment is simple, reagents are safe and non-toxic, the safety of the detection personnel is high, a result is accurate and stable, and damages to instruments are reduced.
Description
Technical field
The present invention relates to the detection of feed addictive composition, specifically refer to total propionic acid content detection method in a kind of mixed feed adjuvant, belong to feed addictive or technical field of analysis and detection.
Background technology
Propionic acid is a kind of volatile fatty acid, is colourless transparent liquid, soluble in water, ethanol, ethyl acetate equal solvent.It is the intermediate product of animal eubolism, has very wide antimicrobial spectrum, is generally used for formula feed mildewproof agent and acidulant.Because the volatility of propionic acid is comparatively strong, add separately propionic acid short for its action time, therefore often add propionate in Midew preventive for feed, propionate common in Midew preventive for feed is propionic acid ammonium, calcium propionate, sodium propionate and potassium propionate simultaneously.Propionate can continue to dissociate propionic acid to play mildew-proof function in acid condition.By a certain amount of propionic acid and salt thereof, and other effective constituents join in the carrier such as silicon dioxide and/or vermiculite to mix and namely can be made into a kind of Midew preventive for feed or acidulant.
The national standard that in Midew preventive for feed, the detection of propionic acid and propionate (in propionic acid) content is ununified at present.The method that feed additive enterprises and feed factory are commonly used has titrimetry, compleximetry, liquid phase chromatography and vapor-phase chromatography etc.First two method is all easily subject to other organic acids impact in mildewproof agent, and therefore accuracy and precision are all lower, can only be used for the bigness scale of propionic acid content.GB/T 17815-1999 defines the method that packed column gas chromatography method measures propionic acid, propionate content in feed, but the method is only applicable to the mensuration of propionic acid content propionic acid, propionate (in propionic acid) below 0.3% in feed, but, in Midew preventive for feed, total propionic acid content is higher, generally be greater than 25%, and at present packed column extensively replace by capillary column, therefore the method is not suitable for the detection of total propionic acid content in Midew preventive for feed.At present, less about using the bibliographical information of total propionic acid content in milk powder Midew preventive for feed.These methods all use excessive mineral acid (sulfuric acid/phosphoric acid) to carry out acidified sample, and mineral acid enters gas chromatography and injection port can be caused to pollute, and serious meeting causes irreversible damage to capillary column.Though adopt headspace sampling to avoid the problems referred to above, but need specific installation, restricted application.Further, in above-mentioned document, pre-treatment step is more, and testing staff's workload is comparatively large, and organic solvent used (as ethyl acetate, acetone, ether etc.) has certain toxicity usually, causes certain threat to the health of testing staff.
Summary of the invention
Loaded down with trivial details for total propionic acid survey timed samples pre-treatment in existing feed addictive, toxic solvent evil, equipment is had to the technical matterss such as infringement, the object of the present invention is to provide a kind of pre-treatment simple to operate, to testing staff's safety non-toxic, and total propionic acid content detection method in the mixed feed adjuvant of instrument damage can be reduced.
To achieve these goals, the technical solution used in the present invention is such:
Total propionic acid content detection method in a kind of mixed feed adjuvant, containing propionic acid and propionate in this mixed feed adjuvant, detecting step is as follows,
1) the corresponding relation typical curve of the propionate concentration that records of making capillary gas chromatography and peak area; Need to configure the propionic acid standard working solution sample introduction of 5-8 concentration gradient between 0.1 ~ 20 mg/mL during step 1) production standard curve to analyze, according to each concentration survey propionic acid peak area the corresponding relation of size and concentration do typical curve, require that related coefficient is greater than 0.99.
2) propionate in detected sample is changed into free propionic acid, in detected sample, add the free propionic acid that enough absolute ethyl alcohols make the propionic acid in detected sample and propionate change into again be dissolved in completely in absolute ethyl alcohol, to be extracted from detected sample by propionic acid, obtain mixed liquor; Add when absolute ethyl alcohol extracts and answer shake well or low temperature ultrasonic, then centrifugal or placement more than 6 hours;
3) the 2nd is got) supernatant of step gained mixed liquor cross disposable organic filter membrane, the peak area of propionic acid in filtrate is recorded by capillary gas chromatography;
4) according to the 1st) the corresponding relation typical curve of propionate concentration and peak area that makes of step, then in conjunction with the 3rd) peak area of propionic acid in the filtrate that obtains of step, in calculating filtrate, the concentration of propionic acid, calculates total propionic acid content in detected sample thus.
Total propionic acid content detection method in a kind of mixed feed adjuvant, containing propionic acid and propionate in this mixed feed adjuvant, detecting step is as follows,
1) the propionic acid standard reserving solution of concentration is determined in preparation, gets this standard reserving solution of certain volume in volumetric flask, then adds the pre-configured internal standard compound standard solution of certain volume, use absolute ethyl alcohol constant volume, obtain propionic acid standard solution; Propionic acid standard solution crosses disposable organic filter membrane, and filtrate with the peak area of capillary gas chromatography propionic acid and internal standard compound, then to calculate under this method propionic acid relative to the correction factor of internal standard compound in conjunction with the concentration of propionic acid standard reserving solution and internal standard compound;
2) propionate in detected sample is changed into free propionic acid, in detected sample, add the free propionic acid that enough absolute ethyl alcohols make the propionic acid in detected sample and propionate change into again be dissolved in completely in absolute ethyl alcohol, to be extracted from detected sample by propionic acid, obtain mixed liquor; Add when absolute ethyl alcohol extracts and answer shake well or low temperature ultrasonic, then centrifugal or placement more than 6 hours;
3) the 2nd is got) supernatant of step gained mixed liquor, this supernatant is considered as the propionic acid standard reserving solution in step 1), by the peak area of step 1) time-and-motion study propionic acid and internal standard compound;
4) according to the 1st) propionic acid that obtains of step relative to the correction factor of internal standard compound, then in conjunction with the 3rd) peak area of propionic acid and internal standard compound in the filtrate that obtains of step, calculate the concentration of propionic acid in filtrate, calculate total propionic acid content in detected sample thus.
Described internal standard compound is crotonic acid standard items, and machine on repeated configuration 6 ~ 8 propionic acid standard solution of answering in step 1) measures, and under calculating this method, propionic acid is relative to the average correction factor of crotonic acid.
Described step 2) in extract propionic acid the amount of absolute ethyl alcohol should to meet in supernatant propionate concentration close to propionic acid standard reserving solution concentration simultaneously.
The present invention is used for the mixed feed adjuvant being carrier with silicon dioxide and/or vermiculite, the particularly chromatogram ration analysis method of total propionic acid content in mildewproof agent and/or acidulant, first utilize strong organic acid that propionate is changed into free propionic acid, extract with absolute ethyl alcohol again, directly get supernatant and cross organic filter membrane, finally get filtrate sample detection.
This method pre-treatment is simple, and agents useful for same safety non-toxic, testing staff's security is high, result accurate stable, and can reduce the damage to instrument.
Accompanying drawing explanation
Propionic acid chromatogram in Fig. 1-mildewproof agent.
Fig. 2-propionic acid typical curve.
Embodiment
Containing propionic acid and propionate in mixed feed adjuvant of the present invention, common propionate is propionic acid ammonium, calcium propionate, sodium propionate and potassium propionate.The present invention has two kinds of methods can realize the detection of total propionic acid content in mixed feed adjuvant, is respectively external standard method and internal standard method.Be described respectively below.
One, external standard method
1) the corresponding relation typical curve of the propionate concentration that records of making capillary gas chromatography and peak area; Need when production standard curve to configure the propionic acid standard working solution sample introduction analysis between 0.1 ~ 20 mg/mL of 5-8 concentration gradient, according to each concentration survey propionic acid peak area the corresponding relation of size and concentration do typical curve, require that related coefficient is greater than 0.99;
2) propionate in detected sample is changed into free propionic acid, in detected sample, add the free propionic acid that enough absolute ethyl alcohols make the propionic acid in detected sample and propionate change into again be dissolved in completely in absolute ethyl alcohol, to be extracted from detected sample by propionic acid, obtain mixed liquor; Add when absolute ethyl alcohol extracts and answer shake well or low temperature ultrasonic, then centrifugal or placement more than 6 hours;
3) the 2nd is got) supernatant of step gained mixed liquor cross disposable organic filter membrane (0.22 μm), the peak area of propionic acid in filtrate is recorded by capillary gas chromatography;
4) according to the 1st) the corresponding relation typical curve of propionic acid peak area and concentration that makes of step, then in conjunction with the 3rd) peak area of propionic acid in the filtrate that obtains of step, in calculating filtrate, the concentration of propionic acid, calculates total propionic acid content in detected sample thus.
In order to improve detection accuracy, the present invention can adopt the method for average, and detected sample is divided into equal several parts, detects by identical operation, the 2nd) the absolute ethyl alcohol amount that adds of step is also equal, finally averages.
Two, internal standard method
1) the propionic acid standard reserving solution of concentration is determined in preparation, gets this standard reserving solution of certain volume in volumetric flask, then adds the pre-configured internal standard compound standard solution of certain volume, use absolute ethyl alcohol constant volume, obtain propionic acid standard solution.Propionic acid standard solution crosses disposable organic filter membrane (0.22 μm), filtrate with the peak area of capillary gas chromatography propionic acid and internal standard compound, then to calculate under this method propionic acid relative to the correction factor of internal standard compound in conjunction with the concentration of propionic acid standard reserving solution and internal standard compound;
2) propionate in detected sample is changed into free propionic acid, in detected sample, add the free propionic acid that enough absolute ethyl alcohols make the propionic acid in detected sample and propionate change into again be dissolved in completely in absolute ethyl alcohol, to be extracted from detected sample by propionic acid, obtain mixed liquor; Add when absolute ethyl alcohol extracts and answer shake well or low temperature ultrasonic, then centrifugal or placement more than 6 hours;
3) the 2nd is got) supernatant of step gained mixed liquor, this supernatant is considered as the propionic acid standard reserving solution in step 1), by the peak area of step 1) time-and-motion study propionic acid and internal standard compound;
4) according to the 1st) propionic acid that obtains of step relative to the correction factor of internal standard compound, then in conjunction with the 3rd) peak area of propionic acid and internal standard compound in the filtrate that obtains of step, calculate the concentration of propionic acid in filtrate, calculate total propionic acid content in detected sample thus.
Internal standard compound should close with boiling point to the relative molecular mass of tested component, structure is similar, and retention time is close, but can separate.The present invention uses crotonic acid, and machine on repeated configuration 6 ~ 8 propionic acid standard solution of answering in step 1) measures, and under calculating this method, propionic acid is relative to the average correction factor of crotonic acid.
Two kinds of method steps 2 above) in the propionate in detected sample changed into free propionic acid carry out as follows, if propionate is unique and be propionic acid ammonium in feed addictive, then without the need to carrying out acidification.Because propionic acid and propionic acid ammonium can be dissolved in absolute ethyl alcohol, during actual treatment, sample first extracts with absolute ethyl alcohol, and propionic acid ammonium is propionic acid and ammonia in gas chromatograph injection port decomposes.If containing other propionates (as calcium propionate, sodium propionate or potassium propionate) in feed addictive, then add formic acid acidifying detected sample, it is that analysis is pure that institute adds formic acid, addition should guarantee all to displace the propionic acid in propionate, formic acid adds and fully to shake up afterwards, and places a period of time.
Above-mentioned steps 2) add absolute ethyl alcohol after answer shake well, the amount of described absolute ethyl alcohol should to meet in supernatant propionate concentration close to propionic acid standard reserving solution concentration.Although the content of propionic acid be can not determine in sample, for Midew preventive for feed and acidulant, its content has an approximate range usually, so can probably estimate in advance, adds absolute ethyl alcohol based on estimation content.Described shake well should avoid the operation being conducive to material volatilization in system.
It is good that vapor-phase chromatography testing conditions should meet propionic acid peak-to-peak shape, and retention time is suitable for.When using internal standard compound, propionic acid peak is separated thoroughly with internal standard compound peak.
Below in conjunction with embodiment, the invention will be further described.
The equipment that the present invention mainly needs has analytical balance, gas chromatograph and support equipment thereof, and propionic acid standard items, analytically pure absolute ethyl alcohol and formic acid etc., and analytical procedure, method are described below:
In embodiment one, propionic acid and propionic acid ammonium type mildewproof agent, total propionic acid content measures (external standard method)
1 reagent and solution
1.1 absolute ethyl alcohols are analyzed pure
1.2 propionic acid standard items
1.3 propionic acid standard reserving solutions: 50mg/mL ethanol solution.(seal rear 4 DEG C of preservations, 3 months shelf-lifves.）
1.4 propionic acid standard working solution: draw 40 respectively, 20,10,4,2mL propionic acid standard reserving solution in 100mL volumetric flask, with absolute ethyl alcohol constant volume (used time now joins), be mixed with 20,10,5,2 and 1mg/mL propionic acid standard working solution.
2 propionic acid contents measure
2.1 principle
Propionic acid and propionic acid ammonium can be dissolved in absolute ethyl alcohol.Sample absolute ethyl alcohol extracts.Propionic acid ammonium is propionic acid and ammonia in injection port decomposes.Use FID to detect propionic acid peak area, then calculate propionate concentration by typical curve, thus calculate total propionic acid content in mildewproof agent.
2.2 sample extraction
Quick and precisely take about 1g (being accurate to 0.0001g) sample in 50mL tool plug triangular flask, add 25mL absolute ethyl alcohol, manually (at least 6h) is spent the night in centrifugal after violent jolting 5min or placement.Careful Aspirate supernatant crosses 0.22 μm of disposable organic filter membrane, and it is to be measured that filtrate is collected in upper machine in sample bottle.
2.3 instruments and condition
2.3.1 instrument
Gas chromatograph: with fid detector;
Chromatographic column: HP-FFAP capillary chromatographic column, 30m × 250 μm × 0.25 μm;
2.3.2 key instrument condition
Adjustment instrument parameter, makes retained fraction be separated with reagent and assorted peak.Design parameter need be made adjustment according to different gas chromatograph.The instrument condition recommended following (being applicable to Agilent 7890B):
Injector temperature: 230 DEG C.
Detector temperature: 230 DEG C.
Carrier gas: nitrogen; Constant current mode, post flow 1.0mL/min.
Post case temperature: initial temperature 100 DEG C, constant temperature 5min.Again with 100 DEG C/min ramp to 200 DEG C.
Sample size: 1 μ L; Split ratio 50:1;
Air mass flow: 400mL/min; Hydrogen flowing quantity: 40mL/min; Make-up gas flow: 30 mL/min;
The calculating of 2.4 results
2.4.1 the formulation of propionic acid typical curve:
Be mixed with 20,10,5,2 and 1mg/mL propionic acid standard working solution are measured by condition above.Take propionate concentration as horizontal ordinate, peak area is that ordinate does typical curve.Fig. 2 is the propionic acid typical curve that embodiment one obtains.Fig. 1 is propionic acid chromatogram in mildewproof agent.
2.4.2 calculate
In sample, active principle propionic acid content is calculated as follows:
In formula:
---the percentage composition of total propionic acid in sample
---the propionate concentration of the detection solution calculated by typical curve, unit mg/mL;
25---sample extracting solution volume, Unit/mL;
---the quality of sample, unit g.
Measurement result retains 2 significant digits.
2.4.3 repeated
Require that twice parallel determinations relative deviation is less than or equal to 10%.
In embodiment two, compound propionic acid type mildewproof agent, total propionic acid content measures (internal standard method)
1 reagent and solution
1.1 absolute ethyl alcohols are analyzed pure
1.2 propionic acid standard items
1.3 crotonic acid standard items
1.4 propionic acid standard reserving solutions: 50 mg/mL ethanol solutions.(seal rear 4 DEG C of preservations, 3 months shelf-lifves.）
1.5 crotonic acid inner mark solutions: 50 mg/mL ethanol solutions.
1.6 propionic acid standard working solution: draw 1mL propionic acid standard reserving solution in 10mL volumetric flask, add 200 μ L crotonic acid inner mark solutions, with absolute ethyl alcohol constant volume (used time now joins).
2 propionic acid contents measure
2.1 principle
With excessive formic acid acidifying mildewproof agent, propionate is dissociated propionic acid completely, then extracts propionic acid with absolute ethyl alcohol, add internal standard compound, constant volume shakes up, and uses gas chromatograph separation determination, calculates propionic acid content in mildewproof agent by internal standard compound.
2.2 sample extraction
Quick and precisely taking 1g(and be accurate to 0.0001g) sample is in 50mL tool plug triangular flask, accurately add 10mL formic acid and absolute ethyl alcohol mixed solution (formic acid: absolute ethyl alcohol volume ratio is 1:4), manually (at least 6h) is spent the night in centrifugal after violent jolting 5min or placement, Aspirate supernatant 1mL is in 10mL volumetric flask, accurately add 200 μ L crotonic acid inner mark solutions, use absolute ethyl alcohol constant volume.Cross 0.22 μm of disposable organic filter membrane after constant volume, it is to be measured that filtrate is collected in upper machine in sample bottle.
2.3 instruments and condition
With embodiment one;
The calculating of 2.4 results
2.4.1 propionic acid content calculates
The correction factor of the relative internal standard compound crotonic acid of propionic acid in this method sample
calculate by following formula (1):
…………………………（1）
In formula:
---the correction factor of the relative internal standard compound of propionic acid in sample;
---the peak area of internal standard compound crotonic acid in propionic acid standard working solution;
---the mass concentration of propionic acid in propionic acid standard working solution, unit is mg/mL;
---the peak area of propionic acid in propionic acid standard working solution;
---the mass concentration of internal standard compound crotonic acid in propionic acid standard working solution, unit is mg/mL.
In sample, total propionic acid content presses following formula (2) calculating:
…………………………（2）
In formula:
---the percentage composition of total propionic acid in testing sample;
---the correction factor of the relative internal standard compound crotonic acid of propionic acid in testing sample;
---the peak area of propionic acid in testing sample solution;
---the concentration of internal standard compound crotonic acid in testing sample solution;
---constant volume, unit is mL;
10---sample extracting solution volume, Unit/mL;
---the peak area of internal standard compound crotonic acid in testing sample solution;
---the quality of sample, unit is g.
Measurement result retains 2 significant digits.
2.4.2 repeated
Twice parallel determinations result relative deviation is less than or equal to 10%.
What finally illustrate is, above embodiment is only in order to illustrate technical scheme of the present invention and unrestricted, although with reference to preferred embodiment to invention has been detailed description, those of ordinary skill in the art is to be understood that, can modify to technical scheme of the present invention or equivalent replacement, and not departing from aim and the scope of technical solution of the present invention, it all should be encompassed in the middle of right of the present invention.
Claims (5)
1. total propionic acid content detection method in a mixed feed adjuvant, the mixed feed adjuvant of this method is mildewproof agent or acidulant, mildewproof agent or acidulant with silicon dioxide and/or vermiculite for carrier, propionic acid and propionate is contained in mildewproof agent or acidulant, it is characterized in that: detecting step is as follows
1) the corresponding relation typical curve of the propionate concentration that records of making capillary gas chromatography and peak area;
2) propionate in detected sample is changed into free propionic acid, in detected sample, add the free propionic acid that enough absolute ethyl alcohols make the propionic acid in detected sample and propionate change into again be dissolved in completely in absolute ethyl alcohol, to be extracted from detected sample by propionic acid, obtain mixed liquor; Add when absolute ethyl alcohol extracts and answer shake well or low temperature ultrasonic, then centrifugal or placement more than 6 hours;
Propionate in detected sample is changed into free propionic acid carry out as follows, if propionate is unique and be propionic acid ammonium, then without the need to carrying out acidification, it can be decomposed into propionic acid and ammonia under injection port hot environment; Otherwise then need to add formic acid acidifying detected sample, institute adds that formic acid is pure for analyzing, and addition should guarantee all to displace the propionic acid in propionate, formic acid adds and fully to shake up afterwards, and places a period of time;
3) the 2nd is got) supernatant of step gained mixed liquor cross disposable organic filter membrane, the peak area of propionic acid in filtrate is recorded by capillary gas chromatography;
4) according to the 1st) the corresponding relation typical curve of propionate concentration and peak area that makes of step, then in conjunction with the 3rd) peak area of propionic acid in the filtrate that obtains of step, in calculating filtrate, the concentration of propionic acid, calculates total propionic acid content in detected sample thus.
2. total propionic acid content detection method in mixed feed adjuvant according to claim 1, it is characterized in that: need to configure the propionic acid standard working solution sample introduction of 5-8 concentration gradient between 0.1 ~ 20 mg/mL during described step 1) production standard curve and analyze, according to each concentration survey propionic acid peak area the corresponding relation of size and concentration do typical curve, require that related coefficient is greater than 0.99.
3. total propionic acid content detection method in a mixed feed adjuvant, the mixed feed adjuvant of this method is mildewproof agent or acidulant, mildewproof agent or acidulant with silicon dioxide and/or vermiculite for carrier, propionic acid and propionate is contained in mildewproof agent or acidulant, it is characterized in that: detecting step is as follows
1) the propionic acid standard reserving solution of concentration is determined in preparation, gets this standard reserving solution of certain volume in volumetric flask, then adds the pre-configured internal standard compound standard solution of certain volume, use absolute ethyl alcohol constant volume, obtain propionic acid standard solution; Propionic acid standard solution crosses disposable organic filter membrane, and filtrate with the peak area of capillary gas chromatography propionic acid and internal standard compound, then to calculate under this method propionic acid relative to the correction factor of internal standard compound in conjunction with the concentration of propionic acid standard reserving solution and internal standard compound;
2) propionate in detected sample is changed into free propionic acid, in detected sample, add the free propionic acid that enough absolute ethyl alcohols make the propionic acid in detected sample and propionate change into again be dissolved in completely in absolute ethyl alcohol, to be extracted from detected sample by propionic acid, obtain mixed liquor; Add when absolute ethyl alcohol extracts and answer shake well or low temperature ultrasonic, then centrifugal or placement more than 6 hours;
Propionate in detected sample is changed into free propionic acid carry out as follows, if propionate is unique and be propionic acid ammonium in feed addictive, then without the need to carrying out acidification, it can be decomposed into propionic acid and ammonia under injection port hot environment; Otherwise then need to add formic acid acidifying detected sample, institute adds that formic acid is pure for analyzing, and addition should guarantee all to displace the propionic acid in propionate, formic acid adds and fully to shake up afterwards, and places a period of time;
3) the 2nd is got) supernatant of step gained mixed liquor, this supernatant is considered as the propionic acid standard reserving solution in step 1), by the peak area of step 1) time-and-motion study propionic acid and internal standard compound;
4) according to the 1st) propionic acid that obtains of step relative to the correction factor of internal standard compound, then in conjunction with the 3rd) peak area of propionic acid and internal standard compound in the filtrate that obtains of step, calculate the concentration of propionic acid in filtrate, calculate total propionic acid content in detected sample thus.
4. total propionic acid content detection method in mixed feed adjuvant according to claim 3, it is characterized in that: described internal standard compound is crotonic acid standard items, machine on repeated configuration 6 ~ 8 propionic acid standard solution of answering in step 1) measures, and under calculating this method, propionic acid is relative to the average correction factor of crotonic acid.
5. total propionic acid content detection method in mixed feed adjuvant according to claim 3, is characterized in that: described step 2) in extract propionic acid the amount of absolute ethyl alcohol should to meet in supernatant propionate concentration close to propionic acid standard reserving solution concentration simultaneously.
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| DE10134658A1 (en) * | 2001-07-20 | 2003-02-06 | Schmack Biogas Ag | Method for VFA determination in anaerobic fermentations |
| CN101650347A (en) * | 2009-09-15 | 2010-02-17 | 安徽泰格生物技术股份有限公司 | Method for simultaneously measuring contents of multiple organic acids in feeder acidulant |
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