CN104458952A - Method for measuring solvent residue amounts of ethyl acetate and n-butyl alcohol in total flavonoids of herba epimedii - Google Patents
Method for measuring solvent residue amounts of ethyl acetate and n-butyl alcohol in total flavonoids of herba epimedii Download PDFInfo
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- CN104458952A CN104458952A CN201410740031.0A CN201410740031A CN104458952A CN 104458952 A CN104458952 A CN 104458952A CN 201410740031 A CN201410740031 A CN 201410740031A CN 104458952 A CN104458952 A CN 104458952A
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- ethyl acetate
- butyl alcohol
- herba epimedii
- total flavonoids
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Abstract
The invention discloses a method for measuring solvent residue amounts of ethyl acetate and n-butyl alcohol in total flavonoids of herba epimedii. The method comprises the following steps: establishing a gas chromatographic method, measuring the residue amounts of organic solvents ethyl acetate and n-butyl alcohol in total flavonoids of herba epimedii. According to the method, methanol serves as a solvent, and isoamyl alcohol serves as an internal standard. The capillary gas chromatography is adopted, the methanol serves as the solvent, the isoamyl alcohol serves as the internal standard, and the residue amounts of the ethyl acetate and n-butyl alcohol in the total flavonoids of herba epimedii are measured. The result proves that the method is easy and convenient to operate, high in sensitivity and high in accuracy and can serve as a method for measuring the residue amounts of the organic solvents in the extracts of total flavonoids of herba epimedii.
Description
Technical field
The present invention relates to the measuring method of ethyl acetate in a kind of barren wort total chromocor, n-butanol solvent residual quantity, belong to biological medicine detection technique field.
Background technology
Barrenwort is Berberidaceae plant barrenwort Epimediumbrevi-cornuMaxim, Epimedium sagittatum Epimedium.sagittatum(Seb.etZucc.) Maxim, the dry leaf of E. Pubescens Epimedium pu-bescens Maxim or korean epimedium herb Epimedium koreanum Nakai, its principle active component is general flavone, icariin, polysaccharide etc., barren wort total chromocor employs ethyl acetate and normal butyl alcohol two kinds of organic solvents in the technique of extraction purification, for ensureing the security of medication, be necessary ethyl acetate, normal butyl alcohol Determination of Residual Organic Solvents carries out studying and controlling.
Summary of the invention
The object of the invention is: the measuring method providing ethyl acetate in a kind of barren wort total chromocor, n-butanol solvent residual quantity, the method simple possible, accuracy of detection is high, meets concerned countries standard, to overcome the deficiencies in the prior art.
Technical scheme of the present invention
The measuring method of ethyl acetate, n-butanol solvent residual quantity in a kind of barren wort total chromocor, the method sets up vapor-phase chromatography to measure organic solvent ethyl acetate and normal butyl alcohol residual quantity in barren wort total chromocor, be solvent with methyl alcohol in the method, isoamylol is interior mark.
In aforesaid a kind of barren wort total chromocor ethyl acetate, n-butanol solvent residual quantity measuring method in, concrete grammar is for adopting capillary gas chromatography, with SE54 capillary column for chromatographic column, temperature programme, nitrogen is carrier gas, injector temperature is 230 DEG C, and fid detector temperature is 240 DEG C.
Owing to adopting technique scheme, compared with prior art, the present invention adopts capillary gas chromatography, take methyl alcohol as solvent, isoamylol is interior mark, measures the ethyl acetate in barren wort total chromocor and normal butyl alcohol residual quantity, result shows, the method is easy and simple to handle, highly sensitive, and accuracy is good, can be used as the assay method of Determination of Residual Organic Solvents in barren wort total chromocor extract.
Accompanying drawing explanation
Accompanying drawing 1 is the gas chromatogram of barren wort total chromocor at control group solution;
Accompanying drawing 2 is the gas chromatogram of barren wort total chromocor at butanol solution;
Accompanying drawing 3 is the gas chromatogram of barren wort total chromocor at negative control solution;
Accompanying drawing 4 is solvent residual amount measurement result schematic diagram in sample;
Accompanying drawing 5 is normal butyl alcohol recovery experimental result schematic diagram;
Accompanying drawing 6 is solvent residual amount measurement result schematic diagram in sample.
Embodiment
Below in conjunction with accompanying drawing, the present invention is described in further detail, but not as any limitation of the invention.
Embodiments of the invention:
1 instrument and reagent
1.1 instrument
GC-14B gas chromatograph (Japanese Shimadzu), hydrogen flame ionization detector, WML chromatographic work station (Nanning Weil-McLain dragon chromatogram scientific & technical corporation), supersonic wave cleaning machine (250W, 29 ~ 34kHz, Beijing Medical Devices two factory) and microsyringe (Town in Shanghai booth microsyringe factory).
1.2 reagent
Normal butyl alcohol, ethyl acetate, isoamylol are chromatographically pure, and methyl alcohol is pure for analyzing; Barren wort total chromocor (lot number 20100912,20100916,20100921) is provided by analytical chemistry teaching and research room of Guiyang Medical College pharmaceutical college.
2 methods and result
2.1 chromatographic condition
SE-54 capillary column (30m × 0 53mm, 1 μm); Injector temperature 230 DEG C, fid detector temperature 240 DEG C, high pure nitrogen is as carrier gas, and before post, pressure is 25kPa, split ratio 10: 1; Hydrogen 50ml/min, air velocity 450ml/min; Adopt temperature programme, initial temperature 35 DEG C (room temperature less than 35 DEG C is carried out) maintains 3min, with 2 DEG C/min ramp to 50 DEG C, maintain 2min, sample size 1.0 μ l, theoretical cam curve is not less than 3000 by ethyl acetate, is not less than 6000 by normal butyl alcohol.
The preparation of 2.2 solution
2.2.1 standard solution preparation
Draw ethyl acetate, normal butyl alcohol is appropriate, add methyl alcohol to make in every 1ml containing ethyl acetate 0.0803mg, the storing solution of normal butyl alcohol 0.0811mg, draw respectively storing solution 0.2,0.6,1.0,1.2,2.0ml, be placed in 10ml measuring bottle, then draw in inner mark solution 0.1ml to each measuring bottle, add methyl alcohol to scale, shake up, obtain series standard solution.
2.2.2 the preparation of inner mark solution
Take isoamylol 0.0182g, put in 50ml measuring bottle, add methyl alcohol and dissolve and be diluted to scale, shake up and get final product.
2.2.3 the preparation of need testing solution
Precision takes three batches of barren wort total chromocors and is about 30mg, puts in 10ml measuring bottle, and accurate absorption inner mark solution 0.1ml, adds methyl alcohol and dissolve and be diluted to scale, shake up, to obtain final product.
2.2.4 the preparation of negative control solution
Take the barren wort total chromocor 30mg without ethyl acetate, normal butyl alcohol purifying, put in 10ml measuring bottle, add methyl alcohol and be dissolved to scale, shake up and make dissolving, to obtain final product.
2.3 specificity tests
The each 1 μ l of accurate absorption reference substance solution, need testing solution and negative control solution, and according to sample introduction analysis under " 2.1 " chromatographic condition, the results are shown in accompanying drawing 1,2 and 3, show that negative control solution is very low in the interference of 4 ~ 12min place, do not affect the quantitative of object, and internal standard compound is separated ideal with each determinand peak.
2.4 linear relationships are investigated
The each standard solution 1 μ l of accurate absorption respectively, sample introduction is analyzed, record peak area, with the concentration of composition to be measured for horizontal ordinate (X), composition to be measured and internal standard compound peak area ratio are ordinate (Y), drawing standard curve, carry out regretional analysis, obtain regression equation, ethyl acetate: Y=0.0023X+0.0739(r=0.9991,1.606 ~ 16.06mg/L); Normal butyl alcohol: Y=0.0032X+0.0271(r=0.9993,1.622 ~ 16.22mg/L), result shows, each determinand linear relationship is good.
2.5 minimum detectability
Progressively diluted by standard solution, the minimum detectability recording ethyl acetate and normal butyl alcohol in solution is respectively 0.5mg/L and 0.3mg/L, is respectively 0.015% and 0.011%(S/N=3 of need testing solution).
2.6 minimum quantitative limit
Progressively diluted by standard solution, the minimum quantitative limit recording ethyl acetate and normal butyl alcohol in solution is respectively 1.7mg/L and 1.0mg/L, is respectively 0.05% and 0.037%(S/N=10 of need testing solution).
2.7 precision test
Get standard solution (ethyl acetate 4.818mg/L, normal butyl alcohol 4.866mg/L, isoamylol 3.640mg/L) continuous sample introduction 6 times, calculate composition to be measured and interior target peak area ratio, the RSD of result ethyl acetate, normal butyl alcohol is respectively 4.6%, 3.2%.
2.8 test sample stability tests
Accurately weighed barren wort total chromocor 30mg, put in 10ml measuring bottle, accurate absorption inner mark solution 0.1ml, add methyl alcohol and be dissolved to scale, respectively at 0,1,2,5,8h sample introduction analyzes, the RSD of each component concentration of result is respectively ethyl acetate 3.6%, normal butyl alcohol 3.9%, shows that need testing solution is stable in 8h.
2.9 replica test
Get barren wort total chromocor sample to prepare according to need testing solution method, the same form 6 parts of sample introductions measure, bring typical curve into, calculate normal butyl alcohol, ethyl acetate content, the average content of ethyl acetate and normal butyl alcohol is respectively 0.061% and 0.066% in 6 parts as a result, RSD is respectively 3.7% and 4.9%, shows that the method is reproducible.
2.10 average recovery is tested
Accurately weighed barren wort total chromocor 30mg, add reference substance solution (ethyl acetate 4.818mg/L, normal butyl alcohol 4.866mg/L) 1ml, 2ml, 3ml respectively, measuring content by drafting method, calculating the recovery, the results are shown in accompanying drawing 4, accompanying drawing 5.
2.11 sample determination
Get each 30mg of barren wort total chromocor sample of three lot numbers, accurately weighedly to put in 10ml measuring bottle, add inner mark solution 0.1ml, add methanol constant volume, shake up, accurately draw the analysis of 1 μ l sample introduction, the results are shown in accompanying drawing 6.
3. discuss
Because barren wort total chromocor is soluble in methyl alcohol, and methyl alcohol boiling point is little compared with ethyl acetate, is separated good in gas chromatography, so select methyl alcohol to be sample solvent.Vapor-phase chromatography assay has internal standard method and external standard method, and internal standard method is comparatively conventional, can reduce sample size, chromatographic condition and the impact of operation on result in an experiment, and accuracy, degree of accuracy are higher, are more suitable for the assay of microcomponent compared with external standard method.So experiment determines to adopt internal mark method determination target solvent residual quantity.When Selection of internal standard, N is chosen in this research, N dimethyl acetamide, isopropyl alcohol and isoamylol respectively with target solvent simultaneously sample introduction, found that there is interference between isopropyl alcohol and target solvent peak, N, N dimethyl acetamide then has conditions of streaking and appearance time is more late, and isoamylol appearance time and degree of separation are better, therefore, select isoamylol to be inner mark solution, methyl alcohol is that dissolve medium is effective.
Different column temperature has been investigated to ethyl acetate in sample solution in experiment, the impact of normal butyl alcohol and isoamylol appearance time, comprehensive test sensitivity, the factor such as error at measurment and test sample analysis time, this research option program temperature-raising method, initial temperature 35 DEG C maintains 3min(room temperature less than 35 DEG C), with 2 DEG C/min ramp to 50 DEG C, maintain 2min, result is good, dissolvent residual measurement result shows ethyl acetate in three batch samples, normal butyl alcohol remains the limit being all less than organic solvent residual in Chinese Pharmacopoeia 2010 editions two annex VIII P subordinate lists 1, meet States Pharmacopoeia specifications.
Claims (2)
1. the measuring method of ethyl acetate, n-butanol solvent residual quantity in a barren wort total chromocor, it is characterized in that: the method sets up vapor-phase chromatography to measure organic solvent ethyl acetate and normal butyl alcohol residual quantity in barren wort total chromocor, be solvent with methyl alcohol in the method, isoamylol is interior mark.
2. the measuring method of ethyl acetate, n-butanol solvent residual quantity in a kind of barren wort total chromocor according to claim 1, it is characterized in that: concrete grammar is for adopting capillary gas chromatography, with SE54 capillary column for chromatographic column, temperature programme, nitrogen is carrier gas, injector temperature is 230 DEG C, and fid detector temperature is 240 DEG C.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106018631A (en) * | 2016-08-16 | 2016-10-12 | 中国烟草总公司郑州烟草研究院 | Analysis method for solvent residue of extract additive |
| CN107290448A (en) * | 2017-06-01 | 2017-10-24 | 广州博济医药生物技术股份有限公司 | A kind of method of residual solvent in detection memantine |
-
2014
- 2014-12-08 CN CN201410740031.0A patent/CN104458952A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106018631A (en) * | 2016-08-16 | 2016-10-12 | 中国烟草总公司郑州烟草研究院 | Analysis method for solvent residue of extract additive |
| CN106018631B (en) * | 2016-08-16 | 2018-06-15 | 中国烟草总公司郑州烟草研究院 | A kind of analysis method of medicinal extract class additive dissolvent residual |
| CN107290448A (en) * | 2017-06-01 | 2017-10-24 | 广州博济医药生物技术股份有限公司 | A kind of method of residual solvent in detection memantine |
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