The assay method of fat emulsion anisidine value
Technical field
The present invention relates to measure the technical field of anisidine value, particularly relate to a kind of fat emulsion methoxyThe assay method of base aniline value.
Background technology
Intravenous lipid emulsion is mainly by oil phase, as soybean oil, median chain triglyceride oil, phosphatide andThe oil-in-water emulsion that water is made through emulsification. Wherein in oil phase and phospholipid composition, contain unsaturated bond, easilyOxidation conversion becomes peroxide. Anisidine value is that reflection peroxide breakdown becomes one of aldehyde, letonesItem index, its value is high, may have infringement to human liver function.
The assay method of existing fat emulsion anisidine value is with reference to " the Fat Emulsion injection of the Chinese pharmacopoeia committeeLiquid (C14-C24) quality standard " (exposure draft). There is following shortcoming and defect part in the method.
Oil phase in fat emulsion as soybean oil and phosphatide can not withstand high temperatures, especially phosphatide, if at heightUnder temperature condition, operation may make the unsaturated fatty acid ester oxidation in soybean oil and phosphatide accelerate, and causes peroxideCompound increase, and the method adopt 60 DEG C of water-bath rotary evaporation in vacuo remove moisture, temperature too high and rotation steamSend out overlong time and may cause peroxide to continue to be degraded to aldehyde, ketone and little molecule aldehyde, rotate through high-temperature vacuumEvaporation may be removed, and causes the inaccurate of measurement result.
Summary of the invention
The invention provides a kind of assay method of fat emulsion anisidine value, in order to improve fat emulsion firstThe accuracy of measurement of oxygen base aniline value.
The assay method of fat emulsion anisidine value of the present invention, comprising:
Adopt alkali metal salt as demulsifier, and adopt ether, as extractant, fat emulsion carry out extraction placeReason, obtains ether phase and water;
Under lower than 40 DEG C of conditions, ether is carried out to the processing of vacuum rotating evaporate to dryness mutually, obtain residue one;
Adopt acetic acid ethyl dissolution residue one, obtain ethyl acetate phase, under lower than 40 DEG C of conditions to acetic acid secondEster carries out the processing of vacuum rotating evaporate to dryness mutually, obtains residue two as test sample.
In technical solution of the present invention, under lower than 40 DEG C of conditions, carry out rotary evaporation in vacuo, avoid high temperature to drawPlay the oxidation of unsaturated fatty acid ester and the problem of degraded, improve the degree of accuracy of measuring, and in fat emulsionPhosphatide be zwitterionic surfactant and have stronger emulsifiability, and alkali metal salt, as sodium chloride,Potassium chloride, sodium sulphate, potassium sulfate etc. are one of good demulsifier, therefore, can avoid in leaching processThe emulsification of sample and can shorten extraction time, improves detection efficiency; Adopt ether as extractant, due toThe boiling point of ether is extremely low can be removed rapidly, therefore can further shorten extraction time; In addition, byMay bring a small amount of moisture in ether, ether cannot be taken a small amount of water out of in removing fast, therefore againWith revolving after the slightly high acetic acid ethyl dissolution residue one of boiling point to steam, moisture is taken out of.
Preferably, the assay method of described fat emulsion anisidine value, also comprises:
Test sample is dissolved with isooctane, as need testing solution, using isooctane as blank, adoptUV-VIS spectrophotometry is measured the absorbance A of need testing solution at 350nm wavelength place1;
Measure isopyknic need testing solution and isooctane, add respectively isopyknic 0.25% 4-methoxyl groupThe glacial acetic acid solution of aniline, obtains respectively test sample mixed liquor and isooctane mixed liquor, with isooctane mixed liquorFor blank, adopt UV-VIS spectrophotometry to measure test sample mixed liquor at 350nm wavelength placeAbsorbance A2。
Concrete, the assay method of described fat emulsion anisidine value, described assay method comprise asLower step:
Precision measures fat emulsion 10mL(milliliter) be placed in separatory funnel;
In described separatory funnel, add ethanol 5mL, mix, then add alkali metal salt 0.05~0.15g andEther 20mL, violent jolting;
Stratification, is transferred to upper oil phase in flask, and lower floor's water is used extracted with diethyl ether twice again, eachEther 10mL, merges oil phase, obtains ether phase;
Under lower than 60 DEG C of conditions, ether is carried out to the processing of vacuum rotating evaporate to dryness mutually, obtain residue one;
Adopt 10mL acetic acid ethyl dissolution residue one, obtain ethyl acetate phase, right under lower than 60 DEG C of conditionsEthyl acetate carries out the processing of vacuum rotating evaporate to dryness mutually, obtains residue three;
Adopt 10mL acetic acid ethyl dissolution residue three, obtain ethyl acetate phase, right under lower than 60 DEG C of conditionsEthyl acetate carries out the processing of vacuum rotating evaporate to dryness mutually, obtains residue two as test sample;
Test sample is dissolved with isooctane and shifts and put in 25mL volumetric flask and be diluted to scale with isooctane,With anhydrous sodium sulfate dehydration, filter after solution as need testing solution;
Get need testing solution, using isooctane as blank, adopt UV-VIS spectrophotometry to existIt is A that 350nm wavelength place records need testing solution absorbance1;
Precision measures need testing solution 10mL and puts in brown tool plug test tube, and precision adds 0.25% 4-methoxyThe glacial acetic acid solution 2mL of base aniline, the jolting of jumping a queue, lucifuge is placed 10 minutes, obtains test sample mixed liquor;Another precision measures isooctane 10mL and replaces need testing solution, with method operation, obtains isooctane mixed liquor, withIsooctane mixed liquor, as blank, adopts UV-VIS spectrophotometry to record at 350nm wavelength placeThe absorbance of test sample mixed liquor is A2, calculate anisidine value by following calculating formula:
In formula: the sampling amount that V is test sample, Unit/mL; B is the labelled amount of oil phase in need testing solution,The g/mL of unit.
Preferably, to the assay method of above-mentioned any fat emulsion anisidine value, described alkali metal saltFor sodium chloride, potassium chloride, sodium sulphate or potassium sulfate.
Alkali metal salt, as demulsifier, can be sodium chloride, potassium chloride, sodium sulphate, potassium sulfate etc., preferablyAdopt sodium chloride, the present invention does not adopt hydrochloric acid as demulsifier, is the pH value that can change water due to hydrochloric acid,Thereby make the pH value of water below the isoelectric point of phosphatide, therefore, can cause part phosphatide to extract not out,May affect the accuracy of measurement result.
In the assay method of above-mentioned any fat emulsion anisidine value, right under lower than 30 DEG C of conditionsEther carries out the processing of vacuum rotating evaporate to dryness mutually; Under lower than 30 DEG C of conditions, ethyl acetate is carried out to vacuum rotating mutuallyEvaporate to dryness processing.
Preferred, in the assay method of above-mentioned fat emulsion anisidine value, lower than 20 DEG C of barsUnder part, ether is carried out to the processing of vacuum rotating evaporate to dryness mutually; Under lower than 20 DEG C of conditions, ethyl acetate is carried out very mutuallyThe processing of empty rotation evaporate to dryness.
The assay method of above-mentioned fat emulsion anisidine value is applicable to the mensuration of following fat emulsion: nutritionEmulsion injection, Java brucea fruit oil emulsion injection, propofol injection, alprostadil injection, elemene injectionLong-chain fat in liquid, Etomidate fatty emulsion parenteral solution, Etomidate medium and long chain fat emulsion injection, PropofolFat emulsion injection, docetaxel fat emulsion injection, ICZ fat emulsion injection, fish oil Fat Emulsion notePenetrate liquid, compound liposoluble vitamins fat emulsion injection, dexamethasone palmitate fat emulsion injection, perfluorCarbocyclic aliphatic emulsion injection, taxol fat emulsion injection, cyclosporin A fat emulsion injection, ZitromaxElement fat emulsion injection, carbamazepine fat emulsion injection, amphotericin B fat emulsion injection, hydroxylCamptothecine fat emulsion injection, diazepam fat emulsion injection, curcuma zedoary oil fat injection, ligustrazine fatFat emulsion injection, dihydroergotoxine methanesulfonate fat emulsion injection or sevoflurane fat emulsion injection.
Detailed description of the invention
In order to improve the accuracy of measurement of fat emulsion anisidine value, the embodiment of the present invention provides oneThe assay method of fat emulsion anisidine value, in this technical scheme, adopts alkali metal salt as breakdown of emulsionAgent, and adopt ether as extractant, the aldehyde in fat emulsion, ketone to be extracted, at vacuum rotating evaporate to drynessTime, temperature all operates under lower than 40 DEG C of conditions, therefore, avoids high temperature to cause the oxygen of unsaturated fatty acid esterThe problem of changing and degrading, improves the degree of accuracy of measuring. For making the object, technical solutions and advantages of the present invention moreAdd clearly, by the following examples the present invention is described in further detail.
The embodiment of the present invention provides a kind of assay method of fat emulsion anisidine value, comprising:
Adopt alkali metal salt as demulsifier, and adopt ether, as extractant, fat emulsion carry out extraction placeReason, obtains ether phase and water;
Under lower than 40 DEG C of conditions, ether is carried out to the processing of vacuum rotating evaporate to dryness mutually, obtain residue one;
Adopt acetic acid ethyl dissolution residue one, obtain ethyl acetate phase, under lower than 40 DEG C of conditions to acetic acid secondEster carries out the processing of vacuum rotating evaporate to dryness mutually, obtains residue two as test sample.
In embodiments of the present invention, under lower than 40 DEG C of conditions, carry out rotary evaporation in vacuo, avoid high temperature to causeThe oxidation of unsaturated fatty acid ester and the problem of degraded, improve the degree of accuracy of measuring, and in fat emulsionPhosphatide is zwitterionic surfactant and has stronger emulsifiability, and alkali metal salt, as sodium chloride, chlorineChanging potassium, sodium sulphate, potassium sulfate etc. is one of good demulsifier, therefore, can avoid sample in leaching processThe emulsification of product and can shorten extraction time, improves detection efficiency; Adopt ether as extractant, due to secondEther has good dissolubility to the phosphatide in fat emulsion and aldehyde, ketone, and the boiling point utmost point low energy of etherEnough be removed rapidly, therefore can further shorten extraction time; In addition, because ether may be brought into fewWater gaging divides, and ether cannot be taken a small amount of water out of in removing fast, therefore again with the slightly high acetic acid of boiling pointAfter ethyl ester dissolution residual substance one, revolve to steam moisture is taken out of.
Refer to that lower than 40 DEG C of conditions solvent can revolve at the temperature steaming lower than 40 in vacuum rotary evaporatorDEG C, for example, can be 39 DEG C, 38 DEG C, 35 DEG C, 30 DEG C, 25 DEG C, 20 DEG C, 15 DEG C, 12 DEG C, 10DEG C, 8 DEG C, 5 DEG C or 2 DEG C, revolve steaming efficiency in order to improve, temperature is generally higher than zero degrees celsius.
Preferably, the assay method of described fat emulsion anisidine value, also comprises:
Test sample is dissolved with isooctane, as need testing solution, using isooctane as blank, adoptUV-VIS spectrophotometry is measured the absorbance A of need testing solution at 350nm wavelength place1;
Measure isopyknic need testing solution and isooctane, add respectively isopyknic 0.25% 4-methoxyl groupThe glacial acetic acid solution of aniline, obtains respectively test sample mixed liquor and isooctane mixed liquor, with isooctane mixed liquorFor blank, adopt UV-VIS spectrophotometry to measure test sample mixed liquor at 350nm wavelength placeAbsorbance A2。
Concrete, the assay method of described fat emulsion anisidine value, described assay method comprise asLower step:
Precision measures fat emulsion 10mL and is placed in separatory funnel;
In described separatory funnel, add ethanol 5mL, mix, then add alkali metal salt 0.05~0.15g andEther 20mL, violent jolting;
Stratification, is transferred to upper oil phase in flask, and lower floor's water is used extracted with diethyl ether twice again, eachEther 10mL, merges oil phase, obtains ether phase;
Under lower than 60 DEG C of conditions, ether is carried out to the processing of vacuum rotating evaporate to dryness mutually, obtain residue one;
Adopt 10mL acetic acid ethyl dissolution residue one, obtain ethyl acetate phase, right under lower than 60 DEG C of conditionsEthyl acetate carries out the processing of vacuum rotating evaporate to dryness mutually, obtains residue three;
Adopt 10mL acetic acid ethyl dissolution residue three, obtain ethyl acetate phase, right under lower than 60 DEG C of conditionsEthyl acetate carries out the processing of vacuum rotating evaporate to dryness mutually, obtains residue two as test sample;
Test sample is dissolved with isooctane and shifts and put in 25mL volumetric flask and be diluted to scale with isooctane,With anhydrous sodium sulfate dehydration, filter after solution as need testing solution;
Get need testing solution, using isooctane as blank, adopt UV-VIS spectrophotometry to existIt is A that 350nm wavelength place records need testing solution absorbance1;
Precision measures need testing solution 10mL and puts in brown tool plug test tube, and precision adds 0.25% 4-methoxyThe glacial acetic acid solution 2mL of base aniline, the jolting of jumping a queue, lucifuge is placed 10 minutes, obtains test sample mixed liquor;Another precision measures isooctane 10mL and replaces need testing solution, with method operation, obtains isooctane mixed liquor, withIsooctane mixed liquor, as blank, adopts UV-VIS spectrophotometry to record at 350nm wavelength placeThe absorbance of test sample mixed liquor is A2, calculate anisidine value by following calculating formula:
In formula: the sampling amount that V is test sample, Unit/mL; B is the labelled amount of oil phase in need testing solution,The g/mL of unit.
Soybean oil and phosphatide in fat emulsion do not dissolve in water, but dissolve in ether, therefore, selectEther is as extractant, and because fat emulsion is O/W(Oil/Water, oil-in-water) type emulsion, therefore,Add appropriate ethanol oil-in-water system can be destroyed, facilitate ether carrying oily matter in fat emulsionGet.
Preferably, to the assay method of above-mentioned any fat emulsion anisidine value, described alkali metal saltFor sodium chloride, potassium chloride, sodium sulphate or potassium sulfate.
Alkali metal salt, as demulsifier, can be sodium chloride, potassium chloride, sodium sulphate, potassium sulfate etc., preferablyAdopt sodium chloride, the present invention does not adopt hydrochloric acid as demulsifier, is the pH value that can change water due to hydrochloric acid,Thereby make the pH value of water below the isoelectric point of phosphatide, therefore, can cause part phosphatide to extract not out,May affect the accuracy of measurement result.
In the assay method of above-mentioned any fat emulsion anisidine value, right under lower than 30 DEG C of conditionsEther carries out the processing of vacuum rotating evaporate to dryness mutually; Under lower than 30 DEG C of conditions, ethyl acetate is carried out to vacuum rotating mutuallyEvaporate to dryness processing.
Experiment showed, 10 DEG C of the every raisings of temperature of revolving steaming, the degradation rate of compound can improve 2~4 times, because ofThis, assay method of the present invention is 40 DEG C of following operations, preferably below 30 DEG C, more preferably below 20 DEG C,Can ensure that peroxide is non-degradable.
Preferred, in the assay method of above-mentioned fat emulsion anisidine value, lower than 20 DEG C of barsUnder part, ether is carried out to the processing of vacuum rotating evaporate to dryness mutually; Under lower than 20 DEG C of conditions, ethyl acetate is carried out very mutuallyThe processing of empty rotation evaporate to dryness.
The assay method of above-mentioned fat emulsion anisidine value is applicable to the mensuration of following fat emulsion: nutritionEmulsion injection or main ingredient be the medicine carrying fat emulsion without UV absorption at 350nm wavelength place, and described main ingredient exists350nm wavelength place comprises Java brucea fruit oil emulsion injection, propofol Injection without the medicine carrying fat emulsion of UV absorptionLong in liquid, alprostadil injection, elemene injection, Etomidate fatty emulsion parenteral solution, EtomidateChain fatty emulsion injection, Propofol medium and long chain fat emulsion injection, docetaxel fat emulsion injection, according to bentHealth azoles fat emulsion injection, fish oil fat emulsion injection, compound liposoluble vitamins fat emulsion injection, palm fibrePalmitic acid acid carulon fat emulsion parenteral solution, perfluorocarbon fat emulsion injection, taxol fat emulsion injection, ringSpore rhzomorph A fat emulsion injection, azithromycin fat emulsion injection, carbamazepine fat emulsion injection,Amphotericin B fat emulsion injection, HCPT fat emulsion injection, diazepam fat emulsion injection,Curcuma zedoary oil fat injection, ligustrazine fat emulsion injection, dihydroergotoxine methanesulfonate fat emulsion injectionOr sevoflurane fat emulsion injection.
Below enumerate the mensuration side that embodiment preferably illustrates fat emulsion anisidine value of the present inventionMethod. But the present invention is not limited to following embodiment.
Embodiment 1
The assay method of fat emulsion anisidine value, step is as follows:
Precision measures fat emulsion (Etomidate fatty emulsion parenteral solution) 10mL and is placed in separatory funnel;
In described separatory funnel, add ethanol 5mL, mix, then add sodium chloride 0.1g and ether20mL, violent jolting;
Stratification, is transferred to upper oil phase in the flask of 250mL, and lower floor's water is used extracted with diethyl ether againTwice, each ether 10mL, merges oil phase, obtains ether phase;
Under lower than 40 DEG C of conditions, ether is carried out to the processing of vacuum rotating evaporate to dryness mutually, obtain residue one;
Adopt 10mL acetic acid ethyl dissolution residue one, obtain ethyl acetate phase, right under lower than 40 DEG C of conditionsEthyl acetate carries out the processing of vacuum rotating evaporate to dryness mutually, obtains residue three;
Adopt 10mL acetic acid ethyl dissolution residue three, obtain ethyl acetate phase, right under lower than 40 DEG C of conditionsEthyl acetate carries out the processing of vacuum rotating evaporate to dryness mutually, obtains residue two as test sample;
Test sample is dissolved with isooctane and shifts and put in 25mL volumetric flask and be diluted to scale with isooctane,With anhydrous sodium sulfate dehydration, filter after solution as need testing solution;
Get need testing solution, using isooctane as blank, adopt UV-VIS spectrophotometry to existIt is A that 350nm wavelength place records need testing solution absorbance1;
Precision measures need testing solution 10mL and puts in brown tool plug test tube, and precision adds 0.25% 4-methoxyThe glacial acetic acid solution 2mL of base aniline, the jolting of jumping a queue, lucifuge is placed 10 minutes, obtains test sample mixed liquor;Another precision measures isooctane 10mL and replaces need testing solution, with method operation, obtains isooctane mixed liquor, withIsooctane mixed liquor, as blank, adopts UV-VIS spectrophotometry to record at 350nm wavelength placeThe absorbance of test sample mixed liquor is A2, calculate anisidine value by following calculating formula:
In formula: the sampling amount that V is test sample, Unit/mL; B is the labelled amount of oil phase in need testing solution,The g/mL of unit; Coefficient 1.2 is: add the solution dilution factor after the glacial acetic acid solution of 4-aminoanisole.
The result of simultaneously measuring 6 duplicate samples is respectively: 1.16,1.24,1.16,1.07,1.14,1.28.
Embodiment 2~6
Embodiment 2~6 adopts embodiment 1 fat emulsion used, only Fat Emulsion in initially adding separatory funnelThe consumption of agent, demulsifier, extractant and ethanol is different, consumption as shown in table 1, and those skilled in the art canThe consumption that steams agent to revolving according to the general knowledge of this area is done corresponding adjusting, and operating procedure is identical with embodiment 1.
In table 1 embodiment 2~6, fat emulsion, demulsifier, extractant and ethanol uses scale
Consumption in embodiment |
Embodiment 2 |
Embodiment 3 |
Embodiment 4 |
Embodiment 5 |
Embodiment 6 |
Fat emulsion (mL) |
5 |
8 |
15 |
20 |
25 |
Sodium chloride (g) |
0.05 |
0.08 |
0.1 |
0.1 |
0.15 |
Ether (mL) |
10 |
20 |
20 |
20 |
20 |
Ethanol (mL) |
2 |
2 |
5 |
5 |
5 |
Anisidine value (mean value) |
1.25 |
1.32 |
1.30 |
1.20 |
1.30 |
Adopt assay method of the present invention to measure the anisidine value of the fat emulsion of different volumes, as calculatedLearn volume (V) and the 1.2A of the fat emulsion measuring2-A1Linear, coefficient R2Reach0.993, therefore, no matter adopt the anisidine value phase of the fat emulsion that the fat emulsion of how many volumes recordsWith, illustrate that assay method of the present invention is workable, and reappearance is better.
Embodiment 7
Embodiment 7 is only different from embodiment 1 operating temperature, and all the other are all identical with embodiment 1, embodiment 71.22,1.17,1.19 under lower than 20 DEG C of conditions, operate, the result of simultaneously measuring 6 duplicate samples is respectively:,1.25、1.20、1.15。
Embodiment 8
Embodiment 8 is only different from embodiment 1 fat emulsion used, and all the other are all identical with embodiment 1, implementsExample 8 fat emulsion used is Propofol medium and long chain fat emulsion injection, measures the result of 6 duplicate samples simultaneously and dividesBe not: 2.01,2.15,2.08,1.98,2.04,1.96.
Embodiment 9
Embodiment 9 is that from the difference of embodiment 8 demulsifier used is different, all the other all with embodiment 8 phasesWith, embodiment 9 demulsifier used is sodium sulphate, the result of simultaneously measuring 6 duplicate samples is respectively: 1.31,1.25、1.19、1.27、1.16、1.22。
Embodiment 10
Embodiment 10 and the difference of embodiment 1 are that fat emulsion used is different with demulsifier used,All the other are all identical with embodiment 1, and embodiment 10 fat emulsion used is nutrition emulsion injection (Fat EmulsionParenteral solution (C14-C24)), demulsifier is potassium sulfate, the result of simultaneously measuring 6 duplicate samples is respectively: 2.10,1.95、1.87、2.07、2.15、1.94。
Embodiment 11
Embodiment 11 and the difference of embodiment 1 are that fat emulsion used is different with demulsifier used,All the other are all identical with embodiment 1, and embodiment 11 fat emulsion used is long chain fat emulsion note in Propofol1.99,2.18,2.06 penetrate liquid, demulsifier is potassium chloride, and the result of simultaneously measuring 6 duplicate samples is respectively:,1.94、2.11、2.02。
Taking soybean oil and phosphatide as the degree of accuracy of blank determination assay method of the present invention, comprise following step respectivelyRapid:
Get soybean oil and within 12 minutes, obtain oil phase test liquid in 121 DEG C of heating, get 1g oil phase test liquid and measure itAnisidine value Ad;
Get lecithin and be dissolved in isooctane and within 12 minutes, obtain phosphatide phase test liquid in 121 DEG C of heating, get 1mLPhosphatide phase test liquid is measured its anisidine value Ap;
Remove the negative sample 10mL of soybean oil, phosphatide, add 1g oil phase test liquid, by embodiment's 1The anisidine value of 6 duplicate samples the A ' that averages are measured in method operation simultaneouslyd;
Remove the negative sample 10mL of soybean oil, phosphatide, add 1mL phosphatide phase test liquid, by embodimentThe anisidine value of 6 duplicate samples the A ' that averages are measured in 1 method operation simultaneouslyp;
The result of respectively method by embodiment 1 being measured with directly get oil phase test liquid or phosphatide phase test liquidMeasurement result comparison, calculate according to following formula:
The rate of recovery that calculates soybean oil and phosphatide is respectively 97.8%, 96.4%, relative standard deviation(RelativeStandardDeviation is called for short RSD) is respectively 3.9%, 3.0%. Illustrate that the present invention surveysThe degree of accuracy of determining method is higher.
Comparative example
Precision measures Etomidate fatty emulsion parenteral solution 10mL and is placed in separatory funnel, adds the salt of 2mol/LAcid 1mL, and add ethanol 1mL along bottle wall, ethyl acetate 10mL, violent jolting. Stratification, by upperLayer solution is transferred in 250mL flask, and water layer is extracted with ethyl acetate twice, and each 10mL, merges secondAcetoacetic ester liquid. With Rotary Evaporators at 40 DEG C of following concentrated solvents that eliminate. Residue adds isooctane and dissolves and shiftPut in 25mL measuring bottle, all the other steps are all identical with embodiment 1. Measure 6 duplicate samples simultaneously.
In comparative example, adopt hydrochloric acid as demulsifier, adopt ethyl acetate as extractant, all the other are usedThe consumption of medicine and medicine is according to consumption shown in embodiment 1, and operating procedure is identical with embodiment 1.The result of simultaneously measuring 6 duplicate samples is respectively: 1.21,1.16,1.18,1.23,1.12,1.14.
In embodiment 1,6 duplicate samples duration used in the time that extracting operation all completes is respectively 3 minutes, 3 pointsClock, 3 minutes, 3 minutes, 3 minutes, 3 minutes, average duration is 3 minutes; And 6 increments in comparative exampleProduct duration used in the time that extracting operation all completes be respectively 10 minutes, 10 minutes, 10 minutes, 10 minutes,10 minutes, 10 minutes, average duration was 10 minutes.
Visible, the relatively shorter and phosphatide of assay method extraction time of the present invention can all be proposed, and greatly carriesThe degree of accuracy of high determination efficiency and mensuration. Comparative example assay method extraction time used length may be byPhosphatide can not be extracted completely in ethyl acetate, and phosphatide is water insoluble, cause phosphatide at water andThe separation of ethyl acetate phase is slower, and adopts ether as extractant, and ether can extract phosphatide completelyCome, therefore the stratification time is shorter.
Reference example
Get fat emulsion sample and add ethanol,aldehyde free and revolve and steam to dry residue and distillate in 60 DEG C, toward distillateMiddle hydro-oxidation potassium test solution is placed 24 hours in 50~60 DEG C of water-baths; Separately, get respectively ethanol,aldehyde free and secondAlcohol (all containing a small amount of aldehyde in 95% ethanol), with method operation, obtains respectively ethanol,aldehyde free liquid and ethanol.
Found that, distillate and ethanol all become yellow, and ethanol,aldehyde free liquid color is constant. Therefore, exist60 DEG C revolve while steaming and also the aldehyde of part can be taken out of and caused surveying except the water in fat emulsion can being taken out of whenDetermine result untrue. Therefore, assay method of the present invention under lower than 40 DEG C of conditions operation even lower thanUnder 20 DEG C of conditions, operate, the measurement result of having avoided high-temperature process to cause is inaccurate.
Obviously, those skilled in the art can carry out various changes and modification and not depart from this present inventionBright design and scope. Like this, if of the present invention these amendment and modification belong to the claims in the present invention andWithin the scope of its equivalent technologies, the present invention be also intended to comprise these change and modification interior.