CN103512971A - Quality control method of brucea javanica oil oral dry emulsion - Google Patents
Quality control method of brucea javanica oil oral dry emulsion Download PDFInfo
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Abstract
The invention relates to a detection method of a traditional Chinese medicine preparation, and in particular relates to a method used for detecting components used for controlling quality. The quality control detection method of a brucea javanica oil dry emulsion is a method for detecting contents of oleic acids and linoleic acids in the brucea javanica oil dry emulsion by using a gas chromatographic method. The quality control detection method of the brucea javanica oil dry emulsion comprises the following steps of: observing shape and properties, identifying, checking and testing contents. In order to control the quality of the brucea javanica oil dry emulsion comprehensively and accurately, the gas chromatographic method is used to identify the characteristic spectrum of the brucea javanica oil dry emulsion and/or composition of fatty acids of the brucea javanica oil oral dry emulsion and the contents of the oleic acids and linoleic acids in the brucea javanica oil dry emulsion are detected at the same time. Verified by an analytical method, the precision, the reproducibility, the stability and the recovery rate of the method meet requirements of the Verification Guideline for Traditional Chinese Medicine Quality Standard Analytical Methods which is an appendix of the 2010 version of Chinese pharmacopoeia.
Description
Invention field
The present invention relates to the method for quality control of the oral dried emulsifier of a kind of brucea fruit oil, belong to the technical field of Chinese medicine.
Background of invention
Java brucea main body of oil oleic acid, it can suppress topoisomerase (TOPOII) activity, thereby suppresses the synthetic and growth of cell DNA, the propagation of blocking-up cancer cell.Brucea fruit oil has selectivity to tumour cancer cell, and the film system such as selective destruction cancer cell membrane and mitochondria, makes cancer cell degeneration necrosis, and harmless to normal cell.Meanwhile, brucea fruit oil has targeting to tumour cancer cell, and after medication, drug concentration is concentrated, and has specificity affinity closely with cancer cell, and humoral immunity and cellular immunity are had to facilitation, and has the hematopoiesis function effect that promotes stem cell.Brucea fruit oil oral agents is widely used in after radiotherapy, chemotherapy auxiliary curing clinically or combined treatment has good curative effect, not only can improve result for the treatment of, also there is increasing leukocyte, alleviate the curative effects such as toxic and side effect, gynecological tumor, breast cancer, brain tumor, metastatic encephaloma, sarcoma, lymthoma, cancer of pancreas, prostate cancer etc. are all had to obvious result for the treatment of.At present, the brucea fruit oil oral agents of using clinically mainly contains brucea fruit oil Orally taken emulsion and two kinds of formulations of brucea fruit oil oral soft capsule.Emulsion has advantages of many uniquenesses, for example, can absorb fast, can be used as the good carrier of the medicine of many poorly water-solubles, and medicine carrying amount is high, has certain targeting etc.But also exist poor stability, the oxidizable factors such as causing unstable product quality of becoming sour of grease.Although soft capsule can slow down the oxidative rancidity of grease to a certain extent, during due to long-term taking, the softgel shell digestion power that patient makes gelatin declines, and easily produces belch, poor compliance.Dried emulsifier in the present invention can solve the problems referred to above, thereby has good prospect in medicine.
The quality control of Java brucea oral latex emulsion adopts the health drug standards promulgated by the ministries or commissions of the Central Government (the 19 of Traditional Chinese medicine historical preparation) WS3-B-3646-98, this standard only checks PH and total acid content, though this method can be controlled glycerine three acid contents of brucea fruit oil, but can not control the active component wherein working, oleic acid and linoleic content, thereby also just can not well guarantee the curative effect of product.Though one of Pharmacopoeia of People's Republic of China adopts vapor phase method to analyze the oleic acid content in Java brucea medicinal material, but the mensuration that does not relate to emulsion, its component is not limited, can not well reflects into and be grouped into, thereby just can not control well the component fluctuation of brucea fruit oil yet yet.Because emulsion is in preparation process, emulsifying agent and other auxiliary materials in order to increase its stability, have been added in a large number.Due to the particularly existence of emulsifying agent of auxiliary material, the assay of medicine is caused to certain interference, therefore, must select easy quantitative analysis method accurately to carry out quality control to emulsion.Conventionally adopt suitable solvent that medicine dissolving is extracted, then carry out assay, auxiliary material interference and impact that especially emulsifying agent is measured extractive content must be considered and get rid of to assay method.Because the oil phase in emulsion is wrapped in inner oil phase, must first make emulsion breakdown of emulsion, discharge oil phase, just can extract.The key that emulsion content is measured is that emulsion extracts the interior oily mutually of emulsion after breakdown of emulsion completely, so the accuracy that emulsion is measured depends on Er Ge aspect: i.e. the extraction of breakdown of emulsion and oil phase.Breakdown of emulsion is to make the profit two-phase of emulsion fluid completely separated, and generally its process realizes in two steps, and the first step is flocculation, and the liquid pearl of disperse phase assembles agglomerating.Second step is coalescent, and in group, each drop is merged into large liquid pearl mutually, finally assembles separated.The breakdown of emulsion of emulsion fluid is with stable relevant with many factors, as the character of two-phase content and component proportion, particle size and distribution, temperature, surface tension, interfacial viscosity and interfacial film etc., wherein interfacial tension interfacial film intensity plays vital effect for the formation of emulsion fluid, stable and breakdown of emulsion.The general chemical demulsification method that adopts is combined use physical demulsification method, can obtain good demulsification.Chemical demulsification method has acid adding, adds alkali, salt adding etc.; That physical demulsification method has is ultrasonic, microwave, heating, electricity and membrane demulsification method, freeze-thaw and high speed centrifugation etc.After breakdown of emulsion, must sample extraction is complete, otherwise affect measurement result, but the existence with surface-active emulsion tends to make extract and is difficult to completely.At present, the breakdown of emulsion that emulsion assay adopts and extracting method are mainly that direct organic solvent extracts and electrolyte destroys emulsion methods, as Sun Lingling (Pharmaceutical Analysis magazine 1996,16 (2): 98) when carrying out large-fruit pungent litse fruit oil breast mensuration, adopt and first with sherwood oil and acetone, carry out two step extractions, with the interference except emulsion breaker Fabaceous Lecithin.Because Litsea cubeba oil has certain solubleness in emulsifying agent, and emulsifying agent also has certain solubleness in water, and the existence of emulsifying agent has increased the solubleness of oil phase in water undoubtedly, makes to extract to be difficult to completely.(post column derivatization HPLC method is measured the related substance of Alprostadil emulsions to Li Ping, Chinese Medicine Leader 2010,7 (34): 47) adopt the method for column front derivation to measure Alprostadil emulsions, get rid of the interference of auxiliary material, though method is feasible, but operate too numerous and diversely, be unfavorable for practical application; Chinese patent CN101596161 has introduced a kind of lipoalprostadil method for measuring, adopt cryodesiccated method to go emulsification, make sample become grease, then use organic solvent extracting grease, obtain test liquid, this method is time-consuming, the impact of emulsifying agent in leaching process is not well eliminated simultaneously.Controlled and safe and effective for guaranteeing product quality, the present invention adopts the oral dried emulsifier emulsion droplet of brucea fruit oil particle diameter to control, the fatty acid of the characteristic spectrum of brucea fruit oil dried emulsifier and/or brucea fruit oil dried emulsifier forms the quality of brucea fruit oil dried emulsifier is limited with becoming to be grouped into, adopt total acid content mensuration and oleic acid and linoleic acid content to measure to control content of triglyceride and oleic acid and the linoleic acid content in product simultaneously, particularly when oleic acid and linoleic acid content being measured by gas chromatography, first with inorganic salts, destroy the structure of emulsion, and then with mixed solvent, guarantee brucea fruit oil to propose completely.
Summary of the invention
The object of the present invention is to provide the method for quality control of the oral dried emulsifier of a kind of brucea fruit oil
Method of quality control of the present invention is realized by the following technical solutions:
One, proterties
Get the oral dried emulsifier of brucea fruit oil appropriate, put in glossy paper, the about 5cm2 that tiles, flattens its surface, at bright place, observe, and be white particle; Sample particle or its emulsion adding after water redissolves are pleasantly sweet.
Two, differentiate
Get the oral dried emulsifier 1g of brucea fruit oil, add absolute ethyl alcohol 10ml and dissolve, filter, filtrate is measured according to UV-VIS spectrophotometry (appendix V A of " Chinese Pharmacopoeia " version in 2010), at the wavelength place of 270nm, has absorption maximum.
Three, check
1, content uniformity
Get 10 bags of the oral dried emulsifiers of brucea fruit oil, the weight of accurately weighed every bag of content respectively, every bag of content weight and labelled amount comparison, content uniformity limit must not surpass ± 5.0%, exceed content uniformity limit, should no more than 2 bags, and must not have 1 bag of a times of exceeding content uniformity limit.(appendix IC of " Chinese Pharmacopoeia " version in 2010)
2, granularity
Get 5 bags of the finished products of the oral dried emulsifier unit dose package of brucea fruit oil, weighed weight is placed in medicine sieve and sieves, maintenance level while sieving, and left and right comes and goes and sieves gently 3 minutes, can not and can, by the particle powder summation of sieving for No. five, must not cross 15% by No. 1 sieve.(appendix XI B second method of " Chinese Pharmacopoeia " version in 2010)
3, moisture
According to aquametry (appendix IX H first method of " Chinese Pharmacopoeia " version in 2010), measure, must not cross 6.0%.
4, emulsibility
Get 1 bag of the oral dried emulsifier of brucea fruit oil, add warm water 200ml, stir 5 minutes, should all dissolve, standing, can layering, after jolting, still can be uniformly dispersed, and must not have insoluble impurities.
5, emulsion droplet particle diameter
Get the oral dried emulsifier 1g of brucea fruit oil, add 60 ℃ of warm water 10ml, stir and make all to dissolve, place 10 minutes, stir, sampling and measuring, particle diameter < 10 μ m must not be less than 99%.
6, limit test of microbe
According to " Chinese Pharmacopoeia " version in 2010, an appendix X III C checks, bacterial population must not be crossed 800CFU/g, and fungi count must not be crossed 80CFU/g, and Escherichia coli, the mite that lives must not detect.
7, characteristic spectrum
According to vapor-phase chromatography (appendix VI E of " Chinese Pharmacopoeia " version in 2010), measure
Chromatographic condition and system suitability chemical bonding polyethylene glycol capillary column (CP WAX-52CB, internal diameter 0.32mm, column length 30m, film thickness 0.50 μ m); 205 ℃ of column temperatures, 250 ℃ of injector temperatures, 300 ℃ of detecting devices (FID), carrier gas High Purity Nitrogen, flow velocity 1ml/min, split ratio 5: 1.Theoretical cam curve should be not less than 10000 according to oleic acid calculated by peak area.It is appropriate that the preparation precision of object of reference solution takes methyl oleate reference substance, methyl linoleate reference substance, methyl stearate reference substance, methyl hexadecanoate reference substance, adds normal hexane and make every 1ml containing the solution of 1mg, as object of reference solution.
8, determination method
Get the about 1g of the oral dried emulsifier of brucea fruit oil, accurately weighed, put in test tube, add water 2ml jolting and make dissolve complete, add saturated metabisulfite solution 1ml, jolting.Add isopropyl alcohol: normal hexane: glacial acetic acid (volume ratio 40: 10: 1) 3ml, jolting, standingly make complete layering, draw the superiors' yellow solution and put 25ml measuring bottle, add normal hexane to scale, shake up, the accurate 1ml that draws, another precision takes oleic acid object of reference solution 2ml, put respectively in 10ml tool plug test tube, with nitrogen, dry up, precision adds 0.5mol/L potassium hydroxide methanol solution 2ml respectively, shakes up, put 60 ℃ of water-bath saponification 25min, every vibration in several minutes once, all disappear to oil droplet, let cool.Precision adds 14% boron trifluoride methanol solution 2ml respectively, shakes up, and puts 60 ℃ of water-bath esterification 2min, lets cool; Precision adds normal hexane 2ml respectively, and shake well adds respectively saturated nacl aqueous solution 1ml, and jolting is standing, and the accurate upper solution 1ml that draws, in 10ml volumetric flask, dries up with nitrogen respectively, adds normal hexane appropriate, and constant volume, shakes up.Draw respectively in each 1 μ L inject gas chromatograph of upper solution, measure, record the chromatogram of 20 minutes, obtain.
Test sample characteristic spectrum should have 4 characteristic peaks, wherein 4 peaks are identical with corresponding object of reference peak retention time, with corresponding peak, methyl oleate object of reference peak be S peak, the relative retention time at calculated characteristics peak 1,2,4, its relative retention time should setting ± 5% within.Setting is 0.60 (peak 1), 0.94 (peak 2), 1.00 (peaks 3), 1.12 (peaks 4).
9, the oral dried emulsifier fatty acid of brucea fruit oil composition measuring
Get the oral dried emulsifier 1g of brucea fruit oil, accurately weighed, put in test tube, add water 2ml jolting and make dissolve complete, add saturated metabisulfite solution 1ml, jolting.Add isopropyl alcohol: normal hexane: glacial acetic acid (volume ratio 40: 10: 1) 3ml, jolting, standingly make complete layering, draw the superiors' yellow solution and put 25ml measuring bottle, add normal hexane to scale, shake up, the accurate 1ml that draws, another precision takes oleic acid object of reference solution 2ml, put respectively in 10ml tool plug test tube, with nitrogen, dry up, precision adds 0.5mol/L potassium hydroxide methanol solution 2ml respectively, shakes up, put 60 ℃ of water-bath saponification 25min, every vibration in several minutes once, all disappear to oil droplet, let cool.Precision adds 14% boron trifluoride methanol solution 2ml respectively, shakes up, and puts 60 ℃ of water-bath esterification 2min, lets cool, precision adds normal hexane 2ml respectively, shake well, add respectively saturated nacl aqueous solution 1ml, jolting, standing, the accurate upper solution 1ml that draws is in 10ml volumetric flask respectively, with nitrogen, dry up, add normal hexane appropriate, constant volume, shake up, for need testing solution, with methyl oleate, methyl linoleate reference substance, the hexane solution of methyl stearate and methyl hexadecanoate is contrast, CP-WAX 52CB (30m * 0.32mm * 0.50 μ m) for chromatographic column, injector temperature: 250 ℃, detector temperature: 300 ℃, column temperature: 205 ℃, split ratio: 5: 1, flow rate of carrier gas: 1mlmin
-1, theoretical cam curve is calculated and all should be not less than 10000 by oleic acid and linoleic acid peak.Fatty acid retention time is followed successively by palmitic acid, stearic acid, oleic acid, linoleic acid.Press areas of peak normalization method and calculate, each fatty acid percentage composition is respectively 10%～15%, 4%～7%, 55%～70% and 15%～28%.
Four, total acid content is measured
Get the oral dried emulsifier 22g of brucea fruit oil, accurately weighed, adding water 22ml leaches, add neutral alcohol 20ml and (add before use instructions phenolphthalein solution 1.0ml, with NaOH vs 0.1mol/L, drop to micro-aobvious pink), precision adds 0.5mol/L potassium hydroxide-ethanol solution 25ml, add hot reflux 40 minutes, with neutral alcohol 10ml, rinse condenser pipe, let cool, add 2 of instructions phenolphthalein solutions, with hydrochloric acid vs (0.5mol/L), be titrated to red disappearance, and the result of titration is proofreaied and correct with blank test.Every 1ml hydrochloric acid vs (0.5mol/L) is equivalent to the C of 141.25mg
18h
34o
2.This product total acid content is pressed oleic acid (C
18h
34o
2) meter, must not be less than 8.0%.
Five, oleic acid and linoleic acid content are measured
1, chromatographic condition and system suitability
CP-WAX 52CB capillary column, fid detector, injector temperature is 250 ℃, and column temperature is 205 ℃, and detector temperature is 300 ℃, and sample size is 1 μ l, split ratio is 5: 1, flow rate of carrier gas: 1mlmin
-1, theoretical cam curve is calculated and is not less than 20000 by oleic acid peak.Under above-mentioned chromatographic condition, each methyl oleate, methyl linoleate and phenol benzoate all reach baseline separation.
2, the preparation of reference substance solution
Get oleic acid, linoleic acid reference substance is appropriate, accurately weighed, add normal hexane and make every 1ml containing the solution of 1.00,0.50mg.
Get phenol benzoate reference substance appropriate, accurately weighed, add normal hexane and make every 1ml containing the solution of 1.00mg, as inner mark solution.
The derivatization of fatty acid: precision measures need testing solution 1ml and puts in right amount in 10ml tool plug test tube respectively, dries up with nitrogen, adds 0.5molL
-1potassium hydroxide methanol solution 2ml, close plug, shakes up, and puts saponification 25min in 60 ℃ of water-baths, let cool, add 15% boron trifluoride methanol solution 2ml, close plug, shakes up, put 60 ℃ of water-bath esterification 3min, let cool, precision adds normal hexane 2ml, jolting, adds saturated NaCl solution 1ml, jolting, standingly make layering, precision is got normal hexane layer solution 1ml, with nitrogen, dries up, precision adds inner mark solution 0.7ml, adds normal hexane and makes to dissolve and be diluted to 10ml, obtains.
3, the preparation of need testing solution
(1) investigation of breaking method
The investigation of A, different inorganic salts
Select the inorganic salts that are commonly used to breakdown of emulsion to carry out breakdown of emulsion processing to the oral dried emulsifier of brucea fruit oil, fixed extraction solvent and extracting method, oleic acid and linoleic extraction ratio after more different inorganic salts breakdowns of emulsion.The results are shown in Table 1.
The investigation of table 1 breaking method
B, the screening of extracting solvent species
The kind of extracting solvent affect extraction efficiency, selects can dissolve oleic acid and can extract oleic acid and the linoleic acid in the oral dried emulsifier of brucea fruit oil by the linoleic organic solvent of fine dissolving again, and the impact of comparison different solvents on extraction ratio, the results are shown in Table 2.
The extraction effect of table 2 different organic solvents
| Extract solvent | Sherwood oil | Normal hexane | Isopropyl alcohol | Ethyl acetate |
| Phenomenon | Emulsification is serious | Emulsification is serious | Emulsification is serious | Emulsification is serious |
| Oleic acid extraction ratio | 11.1 | 16.8 | 19.6 | 79.7 |
| Linoleic acid extraction ratio | 9.4 | 15.8 | 19.3 | 80.6 |
The investigation of C, compounded organic solvent different proportion
Experiment finds, the low organic solvent of polarity is as normal hexane, adds the alcohols can fast emulsion breaking in the emulsion layer that sherwood oil etc. form in leaching process, adds easily emulsification again of vortex again after ethanol breakdown of emulsion, adds no longer emulsification of vortex after isopropyl alcohol breakdown of emulsion; Because oleic acid and linoleic acid contain carboxyl, consider to add a small amount of glacial acetic acid to suppress its hydrolysis and produce salt soluble in water.Comprehensively relatively select normal hexane, isopropyl alcohol, the composite extraction solvent of glacial acetic acid further to investigate the impact of three's ratio on extraction effect, the results are shown in Table 3.
The extraction effect of table 3 different proportion organic solvent
The investigation of D, extraction time
Select and extract solvent normal hexane-isopropyl alcohol-glacial acetic acid (10-40-1), investigate the impact of different extraction times on extraction ratio, the results are shown in Table 4.
The extraction effect of the different extraction times of table 4
| |
1 | 2 | 3 | 4 |
| Oleic acid extraction ratio | 92.6 | 98.5 | 99.7 | 99.7 |
| Linoleic acid extraction ratio | 93.7 | 99.7 | 101.6 | 101.1 |
E, the investigation of extracting solvent load
Select difference to measure and extract solvent investigation extraction effect, the results are shown in Table 5.
Table 5 extracts the extraction effect of solvent load
| Extract solvent load | 7ml | 13ml | 20ml |
| Oleic acid extraction ratio | 99.7 | 92.4 | 86.2 |
| Linoleic acid extraction ratio | 101.1 | 93.5 | 87.0 |
(2) need testing solution preparation method's determines
Comprehensive above-mentioned experimental result finally determines that the preparation method of the oral dried emulsifier need testing solution of brucea fruit oil is: get the about 1g of brucea fruit oil dried emulsifier, accurately weighed, put in test tube, add water 2.0ml jolting and make dissolve complete, add saturated metabisulfite solution 1ml, jolting, add isopropyl alcohol-normal hexane-glacial acetic acid solution (40: 10: 1, V/V) 3ml, 20s is extracted in jolting, standing 5min, draws the superiors' yellow solution.Lower floor's solution divides three extractions, add normal hexane 3ml, vortex 15s, adds isopropyl alcohol-normal hexane-glacial acetic acid solution (40: 10: 1 at every turn, V/V) 2ml, jolting 15s, standing 5min, draws upper strata settled solution, before and after merging, No. four extracts are in 25ml measuring bottle, add normal hexane to scale, shake up, obtain.
(3) need testing solution preparation
The solution 1ml that precision measures after breakdown of emulsion puts in 10ml tool plug test tube in right amount, with nitrogen, dries up, and adds 0.5molL
-1potassium hydroxide methanol solution 2ml, close plug, shakes up, and puts saponification 25min in 60 ℃ of water-baths, let cool, add 15% boron trifluoride methanol solution 2ml, close plug, shakes up, put 60 ℃ of water-bath esterification 3min, let cool, precision adds normal hexane 2ml, jolting, adds saturated NaCl solution 1ml, jolting, standingly make layering, precision is got normal hexane layer solution 1ml, with nitrogen, dries up, precision adds inner mark solution 0.7ml, adds normal hexane and makes to dissolve and be diluted to 10ml, obtains.
(4) preparation of blank solution
According to recipe quantity preparation, containing the blank of brucea fruit oil, do not do newborn granule, accurately weighed, put in test tube, add water 1.5ml jolting and make dissolve complete, add saturated metabisulfite solution 1ml, jolting, add isopropyl alcohol-normal hexane-glacial acetic acid solution (40: 10: 1, V/V) 3ml, 20s is extracted in jolting, standing 5min, draws the superiors' yellow solution.Lower floor's solution divides three extractions, adds normal hexane 3ml, vortex 15s at every turn, add isopropyl alcohol-normal hexane-glacial acetic acid solution (40: 10: 1, V/V) 2ml, jolting 15s, standing 5min, draw upper strata settled solution, before and after merging, No. four extracts, in 25ml measuring bottle, add normal hexane to scale, shake up, the solution 1ml that precision measures after breakdown of emulsion puts in 10ml tool plug test tube in right amount, with nitrogen, dries up, and adds 0.5molL
-1potassium hydroxide methanol solution 2ml, close plug, shakes up, and puts saponification 25min in 60 ℃ of water-baths, let cool, add 15% boron trifluoride methanol solution 2ml, close plug, shakes up, put 60 ℃ of water-bath esterification 3min, let cool, precision adds normal hexane 2ml, jolting, adds saturated NaCl solution 1ml, jolting, standingly make layering, precision is got normal hexane layer solution 1ml, with nitrogen, dries up, precision adds inner mark solution 0.7ml, adds normal hexane and makes to dissolve and be diluted to 10ml, obtains.
(5) measure
Accurate reference substance solution and each 1 μ L inject gas chromatograph of need testing solution drawn, measures, and obtains.
Methodological study of the present invention:
A, linear relationship are investigated
Oleic acid, the linoleic acid standard solution 0.2,1,2,3 drawn respectively, 4ml, puts in 10ml tool plug test tube, with nitrogen, dries up, and adds 0.5molL
-1potassium hydroxide methanol solution 2ml, close plug, shakes up, put saponification 25min in 60 ℃ of water-baths, let cool, add 15% boron trifluoride methanol solution 2ml, close plug, shake up, put 60 ℃ of water-bath esterification 3min, let cool, precision adds normal hexane 2ml, jolting, add saturated NaCl solution 1ml, jolting, standingly makes layering, precision is got normal hexane layer solution 1ml, with nitrogen, dry up, precision adds inner mark solution 0.7ml, adds normal hexane and makes to dissolve and be diluted to 10ml, draw respectively above-mentioned solution 1 μ l inject gas chromatograph and measure, record chromatogram.Peak area ratio y with methyl oleate and phenol benzoate
1for ordinate, with oleic acid concentration x
1for horizontal ordinate calculates typical curve equation I, be y
1=13.76x
1+ 0.013 (r=0.9999).With methyl linoleate and phenol benzoate peak area ratio y
1for ordinate, with linoleic acid concentration x
2for horizontal ordinate calculates typical curve equation II, be y
2=13.86x
2+ 0.006 (r=0.9999).
The investigation of detectability and quantitative limit
Each is appropriate to get methyl oleate, methyl linoleate reference substance, accurately weighed, add normal hexane and be made into successively every 1ml containing each three parts of the solution of methyl oleate, methyl linoleate 1.5,1,0.5 μ g, draw respectively 1 μ l inject gas chromatograph, measure, record chromatogram, as signal to noise ratio (S/N ratio) S/N=3, detectability concentration is 0.5 μ gml
-1.As signal to noise ratio (S/N ratio) S/N=10, quantitative limit concentration is 1.5 μ gml
-1.When quantitative limit concentration, the precision of methyl oleate and methyl linoleate reference substance all meets the requirements.
Table 6 oleic acid and linoleic quantitative limit precision
B, precision are investigated
Get respectively with a reference substance solution and need testing solution, continuous sample introduction 6 times, calculates precision with the peak area ratio of methyl oleate and phenol benzoate and the peak area ratio of methyl linoleate and phenol benzoate respectively.
Table 7 Precision Experiment result
C, study on the stability
Get the about 1g of the oral dried emulsifier of brucea fruit oil, accurately weighed, put in test tube, add water 2ml jolting and make dissolve complete, add saturated metabisulfite solution 1ml, jolting, add isopropyl alcohol: normal hexane: glacial acetic acid solution (40: 10: 1, V/V) 3ml, jolting, standingly make complete layering, draw the superiors' yellow solution and put 25ml measuring bottle.Lower floor's solution repeats to extract three times, adds normal hexane 2ml, jolting at every turn, add isopropyl alcohol: normal hexane: glacial acetic acid solution (40: 10: 1, V/V) 2ml, jolting, standing complete layering, absorption upper strata settled solution, the merging extract of making, add normal hexane to scale, shake up, the accurate 1ml that draws, puts in 10ml tool plug test tube, with nitrogen, dry up, add 0.5molL
-1potassium hydroxide methanol solution 2ml, close plug, shakes up, and puts saponification 25min in 60 ℃ of water-baths, let cool, add 15% boron trifluoride methanol solution 2ml, close plug, shakes up, put 60 ℃ of water-bath esterification 3min, let cool, precision adds normal hexane 2ml, jolting, adds saturated NaCl solution 1ml, jolting, standingly make layering, precision is got normal hexane layer solution 1ml, with nitrogen, dries up, precision adds inner mark solution 0.7ml, adds normal hexane and makes to dissolve and be diluted to 10ml, obtains.
Prepare two parts of need testing solutions, portion is transferred and is set to 0 in room temperature, 1,2,4,8,12,24,36h sample introduction is measured, oleic acid and interior mark peak area ratio RSD=1.23%, and linoleic acid and interior mark peak area ratio RSD=1.22%, illustrate that need testing solution room temperature is placed 12h internal stability good.A in refrigerator (20 ℃) preserve 0,1,2,4,6,8,12,24,36h sample introduction measures, oleic acid and interior mark peak area ratio RSD=1.50%, linoleic acid and interior mark peak area ratio RSD=0.99%, illustrate that need testing solution refrigerator (20 ℃) is preserved 36h internal stability good.
Repeatability is investigated
Parallel 6 parts of the oral dried emulsifiers of brucea fruit oil that take, according to above-mentioned preparation method's operation, sample introduction is measured, and records chromatogram.
Table 8 stability experiment result
Table 9 repeated experiment result
D, average recovery are investigated
Get the oral dried emulsifier of brucea fruit oil, in 80%, 100%, 120% ratio, precision adds oleic acid and linoleic acid standard solution respectively, mix, get the about 1g of the oral dried emulsifier of brucea fruit oil, accurately weighed, put in test tube, add water 2ml jolting and make dissolve complete, add saturated metabisulfite solution 1ml, jolting, add isopropyl alcohol: normal hexane: glacial acetic acid solution (40: 10: 1) 3ml, jolting, standingly makes complete layering, draws the superiors' yellow solution and puts 25ml measuring bottle.Lower floor's solution repeats to extract three times, adds normal hexane 2ml, jolting at every turn, add isopropyl alcohol: normal hexane: glacial acetic acid solution (40: 10: 1) 2ml, jolting, standingly makes complete layering, draw upper strata settled solution, merge extract, add normal hexane to scale, shake up, the accurate 1ml that draws, put in 10ml tool plug test tube, with nitrogen, dry up, add 0.5molL
-1potassium hydroxide methanol solution 2ml, close plug, shakes up, and puts saponification 25min in 60 ℃ of water-baths, let cool, add 15% boron trifluoride methanol solution 2ml, close plug, shake up, put 60 ℃ of water-bath esterification 3min, let cool, precision adds normal hexane 2ml, and jolting adds saturated NaCl solution 1ml, jolting, standingly makes layering, and precision is got normal hexane layer solution 1ml, with nitrogen, dry up, precision adds inner mark solution 0.7ml, adds normal hexane and makes to dissolve and be diluted to 10ml., sample introduction is measured, and records chromatogram.
Table 10 average recovery experimental result
Accompanying drawing explanation
Fig. 1 is that (wherein, peak 1: methyl hexadecanoate for the gas chromatogram of the oral dried emulsifier of brucea fruit oil; Peak 2: methyl stearate; Peak 3: methyl oleate; Peak 4: methyl linoleate), for illustrating that the present invention adopts vapor-phase chromatography identification to be subject to assay method and the result of the characteristic spectrum of test product brucea fruit oil dried emulsifier;
Fig. 2 is the canonical plotting of methyl oleate, for illustrating that the present invention is subject to test product brucea fruit oil dried emulsifier methyl oleate linear relationship to investigate result;
Fig. 3 is the canonical plotting of methyl linoleate, for illustrating that the present invention is subject to test product brucea fruit oil dried emulsifier methyl linoleate linear relationship to investigate result.
embodiment
Embodiment mono-
Proterties: sample (the oral dried emulsifier of brucea fruit oil, lower same) is white particle; Taste is sweet.
Differentiate: sample thief 1g, add absolute ethyl alcohol 10ml and dissolve, to filter, filtrate is measured according to UV-VIS spectrophotometry (appendix V A of " Chinese Pharmacopoeia " version in 2010), at the wavelength place of 270nm, has absorption maximum.
Check: 10 bags of content uniformity sample thiefs, the weight of accurately weighed every bag of content respectively, every bag of content weight and labelled amount comparison, content uniformity limit must not surpass ± 5.0%, exceed content uniformity limit, should no more than 2 bags, and must not have 1 bag of a times of exceeding content uniformity limit.(appendix IC of " Chinese Pharmacopoeia " version in 2010)
Granularity: 5 bags of the finished products of sample thief unit dose package, weighed weight is placed in medicine sieve and sieves, maintenance level while sieving, left and right comes and goes and sieves gently 3 minutes, can not and can, by the granular powder art summation of No. five sieves, must not cross 15% by No. 1 sieve.(appendix XI B second method of " Chinese Pharmacopoeia " version in 2010)
Moisture: measure according to aquametry (appendix IX H first method of " Chinese Pharmacopoeia " version in 2010), must not cross 6.0%.1 bag of emulsibility sample thief, adds warm water 200ml, stirs 5 minutes, should all dissolve, standing, can layering, and after jolting, still can be uniformly dispersed, and must not have insoluble impurities.
Emulsion droplet particle diameter: sample thief 1g, add warm water 10ml, stir and make all to dissolve, place 10 minutes, stir, sampling and measuring, particle diameter < 10 μ m must not be less than 99%.
Limit test of microbe: an appendix X III C checks according to " Chinese Pharmacopoeia " version in 2010, and bacterial population must not be crossed 800CFU/g, and fungi count must not be crossed 80CFU/g, Escherichia coli, the mite that lives must not detect.
Characteristic spectrum: measure according to vapor-phase chromatography (appendix VI E of " Chinese Pharmacopoeia " version in 2010)
Chromatographic condition and system suitability: chemical bonding polyethylene glycol capillary column (CP WAX-52CB, internal diameter 0.32mm, column length 30m, film thickness 0.50 μ m); 205 ℃ of column temperatures, 250 ℃ of injector temperatures, 300 ℃ of detecting devices (FID), carrier gas High Purity Nitrogen, flow velocity 1ml/min, split ratio 5: 1.Theoretical cam curve should be not less than 10000 according to oleic acid calculated by peak area.It is appropriate that the preparation precision of object of reference solution takes methyl oleate reference substance, methyl linoleate reference substance, methyl stearate reference substance, methyl hexadecanoate reference substance, adds normal hexane and make every 1ml containing the solution of 1mg, as object of reference solution.
Determination method: get the dry about 1g of newborn granule of brucea fruit oil, accurately weighed, put in test tube, add water 2ml jolting and make dissolve complete, add saturated metabisulfite solution 1ml, jolting.Add isopropyl alcohol: normal hexane: glacial acetic acid (volume ratio 40: 10: 1) 3ml, jolting, standingly make complete layering, draw the superiors' yellow solution and put 25ml measuring bottle, add normal hexane to scale, shake up, the accurate 1ml that draws, another precision takes oleic acid object of reference solution 2ml, put respectively in 10ml tool plug test tube, with nitrogen, dry up, precision adds 0.5mol/L potassium hydroxide methanol solution 2ml respectively, shakes up, put 60 ℃ of water-bath saponification 25min, every vibration in several minutes once, all disappear to oil droplet, let cool.Precision adds 14% boron trifluoride methanol solution 2ml respectively, shakes up, and puts 60 ℃ of water-bath esterification 2min, lets cool; Precision adds normal hexane 2ml respectively, and shake well adds respectively saturated nacl aqueous solution 1ml, and jolting is standing, and the accurate upper solution 1ml that draws, in 10ml volumetric flask, dries up with nitrogen respectively, adds normal hexane appropriate, and constant volume, shakes up.Draw respectively in each 1 μ L inject gas chromatograph of upper solution, measure, record the chromatogram of 20 minutes, obtain.
Test sample characteristic spectrum should have 4 characteristic peaks, wherein 4 peaks are identical with corresponding object of reference peak retention time, with corresponding peak, methyl oleate object of reference peak be S peak, the relative retention time at calculated characteristics peak 1,2,4, its relative retention time should setting ± 5% within.Setting is 0.60 (peak 1), 0.94 (peak 2), 1.00 (peaks 3), 1.12 (peaks 4).As Fig. 1, the corresponding methyl hexadecanoate in peak 1 wherein, the corresponding methyl stearate in peak 2, the corresponding methyl oleate in peak 3, the corresponding methyl linoleate in peak 4.
Assay: total acid content sample thief 22g, accurately weighed, adding water 22ml leaches, add neutral alcohol 20ml and (add before use instructions phenolphthalein solution 1.0ml, with NaOH vs 0.1mol/L, drop to micro-aobvious pink), precision adds 0.5mol/L potassium hydroxide-ethanol solution 25ml, add hot reflux 40 minutes, with neutral alcohol 10ml, rinse condenser pipe, let cool, add 2 of instructions phenolphthalein solutions, with hydrochloric acid vs (0.5mol/L), be titrated to red disappearance, and the result of titration is proofreaied and correct with blank test.Every 1ml hydrochloric acid vs (0.5mol/L) is equivalent to the C of 141.25mg
18h
34o
2.
This product total acid content is pressed oleic acid (C
18h
34o
2) meter, must not be less than 8.0%.
Oleic acid and linoleic acid: according to vapor-phase chromatography (appendix VI E), measure
Chromatographic condition and system suitability: chemical bonding polyethylene glycol capillary column (CP WAX-52CB, internal diameter 0.32mm, column length 30m, film thickness 0.50 μ m); 205 ℃ of column temperatures, 250 ℃ of injector temperatures, 300 ℃ of detecting devices (FID), carrier gas High Purity Nitrogen, flow velocity 1ml/min, split ratio 5: 1.Theoretical cam curve should be not less than 10000 according to oleic acid calculated by peak area.
The preparation of reference substance solution: precision takes oleic acid and linoleic acid reference substance is appropriate, add respectively normal hexane make every 1ml containing the reference substance solution of 1.0mg oleic acid and every 1ml the solution containing 0.5mg linoleic acid reference substance.
The preparation of inner mark solution: it is appropriate that precision takes phenol benzoate, adds normal hexane and makes every 1ml containing the solution of 1.0mg, as inner mark solution.
Determination method: get the dry about 1g of newborn granule of brucea fruit oil, accurately weighed, put in test tube, add water 2ml jolting and make dissolve complete, add saturated metabisulfite solution 1ml, jolting.Add isopropyl alcohol: normal hexane: glacial acetic acid (volume ratio 40: 10: 1) 3ml, jolting, standingly makes complete layering, draws the superiors' yellow solution and puts 25ml measuring bottle.Lower floor's solution repeats to extract 3 times, adds normal hexane 2ml at every turn, and jolting, adds isopropyl alcohol: normal hexane: glacial acetic acid (volume ratio 40: 10: 1) 2ml, jolting, standingly makes complete layering.Draw upper strata settled solution, merge extract, add normal hexane to scale, shake up, the accurate 1ml that draws, another precision measures oleic acid reference substance solution and each 1ml of linoleic acid reference substance solution, split in 10ml tool plug test tube, with nitrogen, dry up, add respectively 0.5mol/L potassium hydroxide methanol solution 2ml, shake up, put 60 ℃ of water-bath saponification 25min, every vibration in several minutes once, all disappear to oil droplet, let cool.Add respectively 14% boron trifluoride methanol solution 2ml, close plug, shakes up, and puts 60 ℃ of water-bath esterification 2min, lets cool; Precision adds normal hexane 2ml respectively, and shake well adds respectively saturated nacl aqueous solution 1ml, jolting, standing, the accurate upper solution 1ml that draws, in 10ml volumetric flask, dries up with nitrogen respectively, precision adds inner mark solution 0.7ml, adds normal hexane and is settled to scale, shakes up.Draw respectively in each 1 μ L inject gas chromatograph of above-mentioned solution, measure, obtain.
The every 1g of this product is containing oleic acid, with (C
18h
34o
2) meter, must not be less than 45mg; Containing linoleic acid (C
18h
32o
2) meter, must not be less than 13mg.
Embodiment bis-
Proterties: sample (the oral dried emulsifier of brucea fruit oil, lower same) is white particle; Taste is sweet.
Differentiate: sample thief 1g, add absolute ethyl alcohol 10ml and dissolve, to filter, filtrate is measured according to UV-VIS spectrophotometry (appendix V A of " Chinese Pharmacopoeia " version in 2010), at the wavelength place of 270nm, has absorption maximum.
Check:
Content uniformity: 10 bags of sample thiefs, the weight of accurately weighed every bag of content respectively, every bag of content weight and labelled amount comparison, content uniformity limit must not surpass ± 5.0%, exceed content uniformity limit, should no more than 2 bags, and must not have 1 bag of a times of exceeding content uniformity limit.(appendix IC of " Chinese Pharmacopoeia " version in 2010)
Granularity: 5 bags of the finished products of sample thief unit dose package, weighed weight is placed in medicine sieve and sieves, maintenance level while sieving, left and right comes and goes and sieves gently 3 minutes, can not and can, by the particle powder summation of No. five sieves, must not cross 15% by No. 1 sieve.(appendix XI B second method of " Chinese Pharmacopoeia " version in 2010)
Moisture: measure according to aquametry (appendix IX H first method of " Chinese Pharmacopoeia " version in 2010), must not cross 6.0%.1 bag of emulsibility sample thief, adds warm water 200ml, stirs 5 minutes, should all dissolve, standing, can layering, and after jolting, still can be uniformly dispersed, and must not have insoluble impurities.
Emulsion droplet particle diameter: sample thief 1g, add warm water 10ml, stir and make all to dissolve, place 10 minutes, stir, sampling and measuring, particle diameter < 10 μ m must not be less than 99%.
Limit test of microbe: an appendix X III C checks according to " Chinese Pharmacopoeia " version in 2010, and bacterial population must not be crossed 800CFU/g, and fungi count must not be crossed 80CFU/g, Escherichia coli, the mite that lives must not detect.
The oral dried emulsifier fatty acid of brucea fruit oil composition measuring:
According to vapor-phase chromatography (appendix VI E of " Chinese Pharmacopoeia " version in 2010), measure
Chromatographic condition and system suitability: chemical bonding polyethylene glycol capillary column (CP WAX-52CB, internal diameter 0.32mm, column length 30m, film thickness 0.50 μ m); 205 ℃ of column temperatures, 250 ℃ of injector temperatures, 300 ℃ of detecting devices (FID), carrier gas High Purity Nitrogen, flow velocity 1ml/min, split ratio 5: 1.Theoretical cam curve should be not less than 10000 according to oleic acid calculated by peak area.It is appropriate that the preparation precision of object of reference solution takes methyl oleate reference substance, methyl linoleate reference substance, methyl stearate reference substance, methyl hexadecanoate reference substance, adds normal hexane and make every 1ml containing the solution of 1mg, as object of reference solution.
Determination method: get the about 1g of the oral dried emulsifier of brucea fruit oil, accurately weighed, put in test tube, add water 2ml jolting and make dissolve complete, add saturated metabisulfite solution 1ml, jolting.Add isopropyl alcohol: normal hexane: glacial acetic acid (volume ratio 40: 10: 1, V/V) 3ml, jolting, standingly make complete layering, draw the superiors' yellow solution and put 25ml measuring bottle, add normal hexane to scale, shake up, the accurate 1ml that draws, another precision takes oleic acid object of reference solution 2ml, puts respectively in 10ml tool plug test tube, with nitrogen, dries up, precision adds 0.5mol/L potassium hydroxide methanol solution 2ml respectively, shake up, put 60 ℃ of water-bath saponification 25min, every vibration in several minutes once, all disappear to oil droplet, let cool.Precision adds 14% boron trifluoride methanol solution 2ml respectively, shakes up, and puts 60 ℃ of water-bath esterification 2min, lets cool, precision adds normal hexane 2ml respectively, shake well, add respectively saturated nacl aqueous solution 1ml, jolting, standing, the accurate upper solution 1ml that draws is in 10ml volumetric flask respectively, with nitrogen, dry up, add normal hexane appropriate, constant volume, shake up, for need testing solution, with methyl oleate, methyl linoleate reference substance, the hexane solution of methyl stearate and methyl hexadecanoate is contrast, CP-WAX 52CB (30m * 0.32mm * 0.50 μ m) for chromatographic column, injector temperature: 250 ℃, detector temperature: 300 ℃, column temperature: 205 ℃, split ratio: 5: 1, flow rate of carrier gas: 1mlmin
-1, theoretical cam curve is calculated and all should be not less than 10000 by oleic acid and linoleic acid peak.Fatty acid retention time is followed successively by palmitic acid, stearic acid, oleic acid, linoleic acid.Press areas of peak normalization method and calculate, each fatty acid percentage composition is respectively 10%～15%, 4%～7%, 55%～70% and 15%～28%.
Assay: total acid content sample thief 22g, accurately weighed, adding water 22ml leaches, add neutral alcohol 20ml and (add before use instructions phenolphthalein solution 1.0ml, with NaOH vs 0.1mol/L, drop to micro-aobvious pink), precision adds 0.5mol/L potassium hydroxide-ethanol solution 25ml, add hot reflux 40 minutes, with neutral alcohol 10ml, rinse condenser pipe, let cool, add 2 of instructions phenolphthalein solutions, with hydrochloric acid vs (0.5mol/L), be titrated to red disappearance, and the result of titration is proofreaied and correct with blank test.Every 1ml hydrochloric acid vs (0.5mol/L) is equivalent to the C of 141.25mg
18h
34o
2.
This product total acid content is pressed oleic acid (C
18h
34o
2) meter, must not be less than 8.0%.
Oleic acid and linoleic acid: according to vapor-phase chromatography (appendix VI E), measure
Chromatographic condition and system suitability: chemical bonding polyethylene glycol capillary column (CP WAX-52CB, internal diameter 0.32mm, column length 30m, film thickness 0.50 μ m); 205 ℃ of column temperatures, 250 ℃ of injector temperatures, 300 ℃ of detecting devices (FID), carrier gas High Purity Nitrogen, flow velocity 1ml/min, split ratio 5: 1.Theoretical cam curve should be not less than 10000 according to oleic acid calculated by peak area.
The preparation of reference substance solution: precision takes oleic acid and linoleic acid reference substance is appropriate, add respectively normal hexane make every 1ml containing the reference substance solution of 1.0mg oleic acid and every 1ml the solution containing 0.5mg linoleic acid reference substance.
The preparation of inner mark solution: it is appropriate that precision takes phenol benzoate, adds normal hexane and makes every 1ml containing the solution of 1.0mg, as inner mark solution.
Determination method:
Get the dry newborn granule 1g of brucea fruit oil, accurately weighed, put in test tube, add water 2ml jolting and make dissolve complete, add saturated metabisulfite solution 1ml, jolting.Add isopropyl alcohol: normal hexane: glacial acetic acid (volume ratio 40: 10: 1) 3ml, jolting, standingly makes complete layering, draws the superiors' yellow solution and puts 25ml measuring bottle.Lower floor's solution repeats to extract 3 times, adds normal hexane 2ml at every turn, and jolting, adds isopropyl alcohol: normal hexane: glacial acetic acid (volume ratio 40: 10: 1) 2ml, jolting, standingly makes complete layering.Draw upper strata settled solution, merge extract, add normal hexane to scale, shake up, the accurate 1ml that draws, another precision measures oleic acid reference substance solution and each 1ml of linoleic acid reference substance solution, split in 10ml tool plug test tube, with nitrogen, dry up, add respectively 0.5mol/L potassium hydroxide methanol solution 2ml, shake up, put 60 ℃ of water-bath saponification 25min, every vibration in several minutes once, all disappear to oil droplet, let cool.Add respectively 14% boron trifluoride methanol solution 2ml, close plug, shakes up, and puts 60 ℃ of water-bath esterification 2min, lets cool; Precision adds normal hexane 2ml respectively, and shake well adds respectively saturated nacl aqueous solution 1ml, jolting, standing, the accurate upper solution 1ml that draws, in 10ml volumetric flask, dries up with nitrogen respectively, precision adds inner mark solution 0.7ml, adds normal hexane and is settled to scale, shakes up.Draw respectively in each 1 μ L inject gas chromatograph of above-mentioned solution, measure, obtain.
The every 1g of this product is containing oleic acid, with (C
18h
34o
2) meter, must not be less than 45mg; Containing linoleic acid (C
18h
32o
2) meter, must not be less than 13mg.
Claims (8)
1. a method of quality control for the oral dried emulsifier of brucea fruit oil, its brucea fruit oil dried emulsifier be with brucea fruit oil and emulsifying agent and auxiliary material through emulsification, be dried the oral dry emulsion formulation of then making; It is characterized in that, the method for quality control of described brucea fruit oil dried emulsifier comprises following full content:
(1) inspection method of the oral dried emulsifier emulsion droplet of brucea fruit oil particle diameter:
Get this product 1g, add 60 ℃ of warm water 10ml, stir and make all to dissolve, place 10 minutes, stir, sampling and measuring, particle diameter < 10 μ m must not be less than 99%;
(2) emulsion droplet particle diameter:
Sample thief 1g, adds 60 ℃ of warm water 10ml, stirs and makes all to dissolve, and places 10 minutes, stir, and sampling and measuring, particle diameter < 10 μ m must not be less than 99%;
(3) the oral dried emulsifier fatty acid of the characteristic spectrum of the oral dried emulsifier of brucea fruit oil and/or brucea fruit oil composition measuring:
The characteristic spectrum of the oral dried emulsifier of brucea fruit oil:
The preparation of object of reference solution, it is appropriate that precision takes methyl oleate reference substance, methyl linoleate reference substance, methyl stearate reference substance, methyl hexadecanoate reference substance, adds normal hexane and make every 1mL containing the solution of 1mg, as object of reference solution.
Get the oral dried emulsifier 1g of brucea fruit oil, accurately weighed, put in test tube, add water 2ml jolting and make dissolve complete, add saturated metabisulfite solution 1ml, jolting, add isopropyl alcohol: normal hexane: glacial acetic acid (volume ratio 40: 10: 1) double solvents 3ml, jolting, standingly make complete layering, draw the superiors' yellow solution and put 25ml measuring bottle, add normal hexane to scale, shake up, the accurate 1ml that draws, another precision takes oleic acid object of reference solution 2ml, put respectively in 10ml tool plug test tube, with nitrogen, dry up, precision adds 0.5mol/L potassium hydroxide methanol solution 2ml respectively, shakes up, put 60 ℃ of water-bath saponification 25min, every vibration in several minutes once, all disappear to oil droplet, let cool, precision adds 14% boron trifluoride methanol solution 2ml respectively, shakes up, and puts 60 ℃ of water-bath esterification 2min, lets cool, precision adds normal hexane 2ml respectively, shake well, add respectively saturated nacl aqueous solution 1ml, jolting, standing, the accurate upper solution 1ml that draws is in 10ml volumetric flask respectively, with nitrogen, dry up, add normal hexane appropriate, constant volume, shake up, for need testing solution, with methyl oleate, methyl linoleate reference substance, the hexane solution of methyl stearate and methyl hexadecanoate is contrast, CP-WAX 52CB (30m * 0.32mm * 0.50 μ m) for chromatographic column, injector temperature: 250 ℃, detector temperature: 300 ℃, column temperature: 205 ℃, split ratio: 5: 1, flow rate of carrier gas: 1mlmin
-1, theoretical cam curve is calculated and all should be not less than 10000 by oleic acid and linoleic acid peak, in test sample chromatogram, characteristic spectrum should have 4 characteristic peaks, and wherein 4 peaks are identical with corresponding object of reference peak retention time, with corresponding peak, methyl oleate object of reference peak be S peak, the relative retention time at calculated characteristics peak 1,2,4, its relative retention time should setting ± 5% within, setting is 0.60 (peak 1), 0.94 (peak 2), 1.00 (peaks 3), 1.12 (peaks 4),
The oral dried emulsifier fatty acid of brucea fruit oil composition measuring:
Get the oral dried emulsifier 1g of brucea fruit oil, accurately weighed, put in test tube, add water 2ml jolting and make dissolve complete, add saturated metabisulfite solution 1ml, jolting, add isopropyl alcohol: normal hexane: glacial acetic acid (volume ratio 40: 10: 1) 3ml, jolting, standingly make complete layering, draw the superiors' yellow solution and put 25ml measuring bottle, add normal hexane to scale, shake up, the accurate 1ml that draws, another precision takes oleic acid object of reference solution 2ml, put respectively in 10ml tool plug test tube, with nitrogen, dry up, precision adds 0.5mol/L potassium hydroxide methanol solution 2ml respectively, shakes up, put 60 ℃ of water-bath saponification 25min, every vibration in several minutes once, all disappear to oil droplet, let cool, precision adds 14% boron trifluoride methanol solution 2ml respectively, shakes up, and puts 60 ℃ of water-bath esterification 2min, lets cool, precision adds normal hexane 2ml respectively, shake well, add respectively saturated nacl aqueous solution 1ml, jolting, standing, the accurate upper solution 1ml that draws is in 10ml volumetric flask respectively, with nitrogen, dry up, add normal hexane appropriate, constant volume, shake up, for need testing solution, with methyl oleate, methyl linoleate reference substance, the hexane solution of methyl stearate and methyl hexadecanoate is contrast, CP-WAX 52CB (30m * 0.32mm * 0.50 μ m) for chromatographic column, injector temperature: 250 ℃, detector temperature: 300 ℃, column temperature: 205 ℃, split ratio: 5: 1, flow rate of carrier gas: 1mlmin
-1, theoretical cam curve is calculated and all should be not less than 10000 by oleic acid and linoleic acid peak,
(4) the oral dried emulsifier total acid content of brucea fruit oil assay:
Get the oral dried emulsifier 22g of brucea fruit oil, accurately weighed, adding water 22ml leaches, add neutral alcohol 20ml and (add before use instructions phenolphthalein solution 1.0ml, with NaOH vs 0.1mol/L, drop to micro-aobvious pink), precision adds 0.5mol/L potassium hydroxide-ethanol solution 25ml, add hot reflux 40 minutes, with neutral alcohol 10ml, rinse condenser pipe, let cool, add 2 of instructions phenolphthalein solutions, with hydrochloric acid vs (0.5mol/L), be titrated to red disappearance, and the result of titration is proofreaied and correct with blank test; Every 1ml hydrochloric acid vs (0.5mol/L) is equivalent to the C of 141.25mg
18h
34o
2;
(5) the oral dried emulsifier oleic acid of brucea fruit oil and linoleic acid content are measured:
Get the dry newborn granule 1g of brucea fruit oil, accurately weighed, put in test tube, add water 2ml jolting and make dissolve complete, add saturated metabisulfite solution 1ml, jolting; Add isopropyl alcohol: normal hexane: glacial acetic acid (volume ratio 40: 10: 1) 3ml, jolting, standingly makes complete layering, draws the superiors' yellow solution and puts 25ml measuring bottle.Lower floor's solution repeats to extract 3 times, adds normal hexane 2ml at every turn, and jolting, adds isopropyl alcohol: normal hexane: glacial acetic acid (volume ratio 40: 10: 1) 2ml, and jolting, standingly makes complete layering; Draw upper strata settled solution, merge extract, add normal hexane to scale, shake up, the accurate 1ml that draws, another precision measures oleic acid reference substance solution and each 1ml of linoleic acid reference substance solution, split in 10ml tool plug test tube, with nitrogen, dry up, add respectively 0.5mol/L potassium hydroxide methanol solution 2ml, shake up, put 60 ℃ of water-bath saponification 25min, every vibration in several minutes once, all disappear to oil droplet, let cool; Add respectively 14% boron trifluoride methanol solution 2ml, close plug, shakes up, and puts 60 ℃ of water-bath esterification 2min, lets cool; Precision adds normal hexane 2ml respectively, and shake well adds respectively saturated nacl aqueous solution 1ml, jolting, standing, the accurate upper solution 1ml that draws, in 10ml volumetric flask, dries up with nitrogen respectively, precision adds inner mark solution 0.7ml, adds normal hexane and is settled to scale, shakes up.
2. according to method described in claim 1, the total acid content of the oral dried emulsifier of brucea fruit oil is pressed oleic acid (C
18h
34o
2) meter, must not be less than 8.0%; Every 1g brucea fruit oil dried emulsifier is containing oleic acid, with (C
18h
34o
2) meter, must not be less than 45mg; Containing linoleic acid (C
18h
32o
2) meter, must not be less than 13mg.
3. according to method described in claim 1, the characteristic spectrum of the oral dried emulsifier of brucea fruit oil, wherein 4 peaks are identical with corresponding object of reference peak retention time, with corresponding peak, methyl oleate object of reference peak be S peak, the relative retention time at calculated characteristics peak 1,2,4, its relative retention time should setting ± 5% within, setting is 0.60 (peak 1), 0.94 (peak 2), 1.00 (peaks 3), 1.12 (peaks 4).
4. according to the oral dried emulsifier fatty acid of method brucea fruit oil described in claim 1, form, fatty acid retention time is followed successively by palmitic acid, stearic acid, oleic acid, linoleic acid.Press areas of peak normalization method and calculate, each fatty acid percentage composition is respectively 10%～15%, 4%～7%, 55%～70% and 15%～28%.
5. according to method described in claim 1, the characteristic spectrum mensuration of the oral dried emulsifier of brucea fruit oil and/or brucea fruit oil dried emulsifier fatty acid composition measuring and the oral dried emulsifier oleic acid of brucea fruit oil and linoleic acid content are measured breaking method used and are: get the oral dried emulsifier of brucea fruit oil, accurately weighed, put in test tube, add water jolting and make dissolve complete, add saturated metabisulfite solution jolting, add isopropyl alcohol-normal hexane-glacial acetic acid solution, 20s is extracted in jolting, standing 5 minutes, draws the superiors' yellow solution; Lower floor's solution extracts three times again, first adds normal hexane jolting 15 seconds at every turn, then adds isopropyl alcohol-normal hexane-glacial acetic acid solution jolting 15 seconds, after standing 5 minutes, draw upper strata settled solution, before and after merging, No. four extracts are in 25ml measuring bottle, add normal hexane to scale, shake up, obtain.
6. according to method described in claim 3, breakdown of emulsion process operates in the following order, first adds saturated inorganic salt solution shake well, then adds normal hexane jolting, finally adds compounded organic solvent and extracts.
7. according to method described in claim 6, the saturated solution that saturated inorganic salt solution is sodium sulphate.
8. according to method described in claim 6, compounded organic solvent is isopropyl alcohol-normal hexane-glacial acetic acid solution, and its proportion of composing is volume ratio (V/V) 40: 10: 1.
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| CN104655746A (en) * | 2015-01-16 | 2015-05-27 | 沈阳药大雷允上药业有限责任公司 | Detection method for medicinal compositions of brucea javanica medicinal material, oleum fructus brucease and oleum fructus brucease pharmaceutical preparation |
| CN104777260A (en) * | 2014-12-26 | 2015-07-15 | 李宏 | Brucea javanica oil emulsion injection liquid quality control method |
| CN106840799A (en) * | 2017-02-27 | 2017-06-13 | 浙江圣兆药物科技股份有限公司 | The assay method of anisidine value in a kind of dried emulsifier |
| CN107091890A (en) * | 2017-04-25 | 2017-08-25 | 浙江圣兆药物科技股份有限公司 | The assay method of oleic acid sodium content in a kind of injection dried emulsifier |
| CN114200028A (en) * | 2020-09-18 | 2022-03-18 | 北京远大九和药业有限公司 | Demulsification method and embedding rate detection method of oral emulsion containing eucalyptol, limonene and alpha-pinene |
| CN115420832A (en) * | 2022-09-20 | 2022-12-02 | 贵州医科大学 | GC-MS fingerprint spectrum and multi-index content determination method of litsea pungens oil |
| CN116068065A (en) * | 2021-10-29 | 2023-05-05 | 中昊晨光化工研究院有限公司 | Method for detecting content of perfluorinated acid compounds in polymer emulsion |
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