CN104730166A - Quantitative determination method for 3,3'-dimethyl ellagic acid and 3,3',4'-trimethyl ellagic acid in polygonum runcinatum - Google Patents
Quantitative determination method for 3,3'-dimethyl ellagic acid and 3,3',4'-trimethyl ellagic acid in polygonum runcinatum Download PDFInfo
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- 229960002852 ellagic acid Drugs 0.000 title claims abstract description 42
- FAARLWTXUUQFSN-UHFFFAOYSA-N methylellagic acid Natural products O1C(=O)C2=CC(O)=C(O)C3=C2C2=C1C(OC)=C(O)C=C2C(=O)O3 FAARLWTXUUQFSN-UHFFFAOYSA-N 0.000 title claims abstract description 42
- AFSDNFLWKVMVRB-UHFFFAOYSA-N Ellagic acid Chemical compound OC1=C(O)C(OC2=O)=C3C4=C2C=C(O)C(O)=C4OC(=O)C3=C1 AFSDNFLWKVMVRB-UHFFFAOYSA-N 0.000 title claims abstract description 41
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- 238000004445 quantitative analysis Methods 0.000 title abstract 4
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- 239000000523 sample Substances 0.000 claims description 41
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- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 33
- 150000007513 acids Chemical class 0.000 claims description 31
- 239000000284 extract Substances 0.000 claims description 20
- 239000013558 reference substance Substances 0.000 claims description 18
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 17
- 239000012488 sample solution Substances 0.000 claims description 15
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- 239000000243 solution Substances 0.000 claims description 12
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- 238000002360 preparation method Methods 0.000 claims description 7
- 239000002904 solvent Substances 0.000 claims description 7
- 239000000047 product Substances 0.000 claims description 6
- UGAPHEBNTGUMBB-UHFFFAOYSA-N acetic acid;ethyl acetate Chemical compound CC(O)=O.CCOC(C)=O UGAPHEBNTGUMBB-UHFFFAOYSA-N 0.000 claims description 5
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- Heterocyclic Carbon Compounds Containing A Hetero Ring Having Oxygen Or Sulfur (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a quantitative determination method for 3,3'-dimethyl ellagic acid and 3,3',4'-trimethyl ellagic acid in polygonum runcinatum. The quantitative determination method comprises the following steps: measuring the peak area by employing a high performance liquid chromatography under a specific chromatographic condition, and then calculating the content of the 3,3'-dimethyl ellagic acid and the 3,3',4'-trimethyl ellagic acid in the polygonum runcinatum through a linear equation. The quantitative determination method has the characteristics that the medicinal material quality of the polygonum runcinatum can be effectively controlled by quantitative determination of the 3,3'-dimethyl ellagic acid and the 3,3',4'-trimethyl ellagic acid in the polygonum runcinatum; and the method is easy to operate, stable in result and high in accuracy.
Description
Technical field
The invention belongs to medicine quantitative measurement technology field, relate in particular to the quantitative detecting method of 3,3'-dimethylellagic acids in a kind of runcinate knotweed herb, 3,3', 4'-trimethyl ellagic acids.
Background technology
3,3'-dimethylellagic acid and 3,3', 4'-trimethyl ellagic acid belongs to the derivant of ellagic acid, extensively be present in plantage, there is stronger anti peroxidation of lipid and the ability of scavenging free radicals, have inhibiting effect to various bacteria and virus, can suppress the canceration that the position such as liver, kidney is brought out due to chemical induction, it also has coagulation function.
Runcinate knotweed herb derives from the dry rhizome of polygonaceae arsesmart runcinate knotweed herb (Polygonum runcinatum Buch.-Ham.var.sinene Hemsl.), another name soil is done one's utmost, gaily-colored butterfly, aleppo dock rhizome, snakehead knotweed etc.Early summer or autumn excavate, cold in nature, bitter, puckery, micro-acid.There is detumescence of activating blood and relaxing tendons, clearing heat and detoxicating effect, and there is good antioxidation activity and to the obvious bacteriostasis of G-Shigella shigae.Cure mainly dysentery, have loose bowels, spit blood spitting of blood, hemorrhoid hemorrhage, leukorrhea with reddish discharge, dysmenorrhoea, venomous snake bite, internal lesion caused by overexertion pain in the back and traumatic injury etc., external application control mastitis, carbuncle furuncle poison etc.
Find in the chemical constitution study of runcinate knotweed herb, the content of its ellagic acid compounds is higher.Therefore, the content of ellagic acid analog derivative in runcinate knotweed herb is the important indicator weighing its drug effect.
Runcinate knotweed herb now records in " Guizhou Province's Chinese crude drug, the Ethnic crude drugs quality standard " of version in 2003, current open source literature only relates to runcinate knotweed herb proterties and discriminating, for the assay of its effective ingredient, at present, only have spectrophotometric method to carry out the bibliographical information of determination of total flavonoids in runcinate knotweed herb, the method measures absorbance according to after colour developing, broadly calculate the content of content as general flavone of substance that show color, at the bottom of specificity and accuracy, measurement result deviation is comparatively large, is unfavorable for Measurement accuracy.
Because ellagic acid compounds has the multiple biologically actives such as anti-oxidant, anti-inflammatory, blood coagulation, anti-cancer, be one of effective substance of runcinate knotweed herb, with it as quantitative target, rationally more scientific, the quality of runcinate knotweed herb medicinal material can be made to obtain more effective control.
Summary of the invention
The object of the invention is to overcome aforementioned drawback and the one provided effectively can control runcinate knotweed herb quality of medicinal material, the quantitative detecting method of 3,3'-dimethylellagic acids, 3,3', 4'-trimethyl ellagic acids in the runcinate knotweed herb that method is easy to operate, result is stablized, precision is high.
The quantitative detecting method of 3,3'-dimethylellagic acids, 3,3', 4'-trimethyl ellagic acids in a kind of runcinate knotweed herb of the present invention, comprises the following steps:
(1) sample solution preparation: get runcinate knotweed herb medicinal material, pulverizes, and crosses 60 mesh sieves, take 1.0g, accurately weighed, be placed in container, add 100ml alcohol reflux and extract 1h, reclaim the solvent in extract and be concentrated into medicinal extract shape, adding 20ml water, ultrasonic process 20min, extracts 5 times with ethyl acetate jolting, and the consumption of ethyl acetate is each 20ml, evaporate to dryness acetic acid ethyl acetate extract, residue adds methyl alcohol and dissolves and be settled in 10ml measuring bottle, shakes up, 0.45 μm of miillpore filter filters, obtained sample solution;
(2) sample size measures:
Chromatographic condition:
C18 chromatographic column;
Mobile phase: volume ratio is the methyl alcohol of 7-27: 20-50: 23-73: acetonitrile: 0.1% phosphoric acid;
Determined wavelength: 230-270nm;
Column temperature: 25-40 DEG C;
Volumetric flow rate: 0.8-1.2mlmin
-1;
Sample size: 10-20 μ l;
With high performance liquid chromatograph, sample solution is detected, according to the content of the peak area recorded by 3,3'-dimethylellagic acids in linear equation calculation sample and 3,3', 4'-trimethyl ellagic acid:
The linear equation of 3,3'-dimethylellagic acid is: Y
1=5598X
1+ 180.8, r=0.9991;
The linear equation of 3,3', 4'-trimethyl ellagic acid is: Y
2=5517.4X
2+ 73.3, r=0.9990;
Wherein, Y
1represent the peak area that 3,3'-dimethylellagic acid utilizes high performance liquid chromatograph to detect, Y
2represent the peak area that 3,3', 4'-trimethyl ellagic acid utilizes high performance liquid chromatograph to detect, X
1represent the sample introduction quality of 3,3'-dimethylellagic acid, X
2represent the sample introduction quality of 3,3', 4'-trimethyl ellagic acid, r is related coefficient.
The quantitative detecting method of 3,3'-dimethylellagic acids, 3,3', 4'-trimethyl ellagic acids in a kind of runcinate knotweed herb of the present invention, wherein: the linear equation of described 3,3'-dimethylellagic acids, 3,3', 4'-trimethyl ellagic acids obtains by the following method:
Get 3,3'-dimethylellagic acid reference substance 19.8mg and 3,3', 4'-trimethyl ellagic acid reference substance 9.82mg, adds methyl alcohol and dissolves and constant volume, makes every 1ml containing 3, the mixing reference substance stock solution of 3'-dimethylellagic acid 0.0990mg and 3,3', 4'-trimethyl ellagic acid 0.0491mg;
Precision measure mixing reference substance stock solution 2,4,6,8mL, put in 10mL measuring bottle, add methanol dilution constant volume, be mixed with gradient solution, make control series product solution together with former mixing reference substance stock solution;
Adopt chromatographic condition: C18 chromatographic column, mobile phase: volume ratio is the methyl alcohol of 7-27: 20-50: 23-73: acetonitrile: 0.1% phosphoric acid, determined wavelength: 230-270nm, column temperature: 25-40 DEG C, volumetric flow rate: 0.8-1.2mlmin
-1, sample size: 10-20 μ l, utilizes high performance liquid chromatograph to detect, with the sample introduction quality of reference substance be horizontal ordinate, peak area for ordinate, will sample introduction quality and the peak area of above-mentioned variable concentrations titer be recorded, by linear regression and get final product.
The quantitative detecting method of 3,3'-dimethylellagic acids, 3,3', 4'-trimethyl ellagic acids in a kind of runcinate knotweed herb of the present invention, wherein: described C18 chromatographic column is Dikma ODS C18 post, and the length of chromatographic column is 250mm, and diameter is 4.6mm, and packing material size is 5 μm.
The quantitative detecting method of 3,3'-dimethylellagic acids, 3,3', 4'-trimethyl ellagic acids in a kind of runcinate knotweed herb of the present invention, wherein: the first of described mobile phase to be volume ratio be 17:33:50: acetonitrile: 0.1% phosphoric acid.
The quantitative detecting method of 3,3'-dimethylellagic acids, 3,3', 4'-trimethyl ellagic acids in a kind of runcinate knotweed herb of the present invention, wherein: the methyl alcohol in described mobile phase and acetonitrile are Chromatographic Pure Methanol and acetonitrile, described phosphoric acid is for analyzing pure phosphoric acid.
The quantitative detecting method of 3,3'-dimethylellagic acids, 3,3', 4'-trimethyl ellagic acids in a kind of runcinate knotweed herb of the present invention, wherein: described determined wavelength is 248nm.
The quantitative detecting method of 3,3'-dimethylellagic acids, 3,3', 4'-trimethyl ellagic acids in a kind of runcinate knotweed herb of the present invention, wherein: described column temperature is 30 DEG C.
The quantitative detecting method of 3,3'-dimethylellagic acids, 3,3', 4'-trimethyl ellagic acids in a kind of runcinate knotweed herb of the present invention, wherein: described volumetric flow rate is 1.0mlmin
-1.
The quantitative detecting method of 3,3'-dimethylellagic acids, 3,3', 4'-trimethyl ellagic acids in a kind of runcinate knotweed herb of the present invention, wherein: described sample size is 10 μ l.
The present invention compared with prior art, there is obvious beneficial effect, as can be known from the above technical solutions: in runcinate knotweed herb provided by the present invention 3,3'-dimethylellagic acid, 3,3', the quantitative detecting method of 4'-trimethyl ellagic acid adopts high performance liquid chromatography, and has made corresponding restriction through great many of experiments to chromatographic condition parameter, solvent etc.Can well by 3 using methanol-acetonitrile-0.1% phosphoric acid as mobile phase, 3'-dimethylellagic acid and 3,3', 4'-trimethyl ellagic acid is separated, and select ethanol as sample extraction solvent, extraction efficiency is high and non-toxic, in Sample Preparation Procedure, adopt extraction into ethyl acetate, avoid impurity and interference is caused to experimental result, improve the precision of detection.Adopt this law to measure method of quality control that 3,3'-dimethylellagic acids in red shin medicinal material and 3,3', 4'-trimethyl ellagic acid content can be used as runcinate knotweed herb, can effectively control runcinate knotweed herb quality of medicinal material.
Embodiment
Below in conjunction with embodiment, beneficial effect of the present invention is described in further detail.
the linear equation of 3,3'-dimethylellagic acid, 3,3', 4'-trimethyl ellagic acids obtains by the following method:
Get 3,3'-dimethylellagic acid reference substance 19.8mg and 3,3'; 4'-trimethyl ellagic acid reference substance 9.82mg, adds methyl alcohol and dissolves and constant volume, makes every 1ml containing 3; the mixing reference substance stock solution of 3'-dimethylellagic acid 0.0990mg and 3,3', 4'-trimethyl ellagic acid 0.0491mg;
Precision measure mixing reference substance stock solution 2,4,6,8mL, put in 10mL measuring bottle, add methanol dilution constant volume, be mixed with gradient solution, make control series product solution together with former mixing reference substance stock solution;
Adopt chromatographic condition: C18 chromatographic column, mobile phase: volume ratio is the methyl alcohol of 7-27: 20-50: 23-73: acetonitrile: 0.1% phosphoric acid, determined wavelength: 230-270nm, column temperature: 25-40 DEG C, volumetric flow rate: 0.8-1.2mlmin
-1, sample size: 10-20 μ l, high performance liquid chromatograph is utilized to detect, 3,3'-dimethylellagic acid and 3,3', 4'-trimethyl ellagic acid has good linear relationship with peak area respectively within the scope of 0.198-0.990 μ g and 0.098-0.491 μ g, will record sample introduction quality and the peak area of above-mentioned variable concentrations titer, and be obtained by linear regression:
The linear equation of 3,3'-dimethylellagic acid is: Y
1=5598X
1+ 180.8, r=0.9991;
The linear equation of 3,3', 4'-trimethyl ellagic acid is: Y
2=5517.4X
2+ 73.3, r=0.9990;
Wherein, Y
1represent the peak area that 3,3'-dimethylellagic acid utilizes high performance liquid chromatograph to detect, Y
2represent the peak area that 3,3', 4'-trimethyl ellagic acid utilizes high performance liquid chromatograph to detect, X
1represent the sample introduction quality of 3,3'-dimethylellagic acid, X
2represent the sample introduction quality of 3,3', 4'-trimethyl ellagic acid, r is related coefficient.
embodiment 1-10
The quantitative detecting method of 3,3'-dimethylellagic acids, 3,3', 4'-trimethyl ellagic acids in a kind of runcinate knotweed herb medicinal material, wherein, runcinate knotweed herb medicinal material is respectively 10 batches that gather autumn, comprises the following steps:
(1) sample solution preparation: get runcinate knotweed herb medicinal material, pulverizes, and crosses 60 mesh sieves, take 1.0g, accurately weighed, be placed in container, add 100ml alcohol reflux and extract 1h, reclaim the solvent in extract and be concentrated into medicinal extract shape, adding 20ml water, ultrasonic process 20min, extracts 5 times with ethyl acetate jolting, and the consumption of ethyl acetate is each 20ml, evaporate to dryness acetic acid ethyl acetate extract, residue adds methyl alcohol and dissolves and be settled in 10ml measuring bottle, shakes up, 0.45 μm of miillpore filter filters, obtained sample solution;
(2) sample size measures:
Chromatographic condition:
C18 chromatographic column, enlightening horse Dikma ODS C18 post, the length of chromatographic column is 250mm, and diameter is 4.6mm, and packing material size is 5 μm;
Mobile phase: volume ratio is the methyl alcohol of 17:33:50: acetonitrile: 0.1% phosphoric acid, wherein, methyl alcohol and acetonitrile are Chromatographic Pure Methanol and acetonitrile, and phosphoric acid is for analyzing pure phosphoric acid;
Determined wavelength: 248nm;
Column temperature: 30 DEG C;
Volumetric flow rate: 1.0mlmin
-1;
Sample size: 10 μ l;
With high performance liquid chromatograph, sample solution is detected, according to the content of the peak area recorded by 3,3'-dimethylellagic acids in linear equation calculation sample and 3,3', 4'-trimethyl ellagic acid.
The assay of 10 batch sample the results are shown in Table 1.
embodiment 11-12
The quantitative detecting method of 3,3'-dimethylellagic acids, 3,3', 4'-trimethyl ellagic acids in runcinate knotweed herb medicinal material, wherein, runcinate knotweed herb medicinal material is respectively 2 batches that gather summer, comprises the following steps:
(1) sample solution preparation: get runcinate knotweed herb medicinal material, pulverizes, and crosses 60 mesh sieves, take 1.0g, accurately weighed, be placed in container, add 100ml alcohol reflux and extract 1h, reclaim the solvent in extract and be concentrated into medicinal extract shape, adding 20ml water, ultrasonic process 20min, extracts 5 times with ethyl acetate jolting, and the consumption of ethyl acetate is each 20ml, evaporate to dryness acetic acid ethyl acetate extract, residue adds methyl alcohol and dissolves and be settled in 10ml measuring bottle, shakes up, 0.45 μm of miillpore filter filters, obtained sample solution;
(2) sample size measures:
Chromatographic condition:
C18 chromatographic column, enlightening horse Dikma ODS C18 post, the length of chromatographic column is 250mm, and diameter is 4.6mm, and packing material size is 5 μm;
Mobile phase: volume ratio is the methyl alcohol of 15:40:45: acetonitrile: 0.1% phosphoric acid;
Determined wavelength: 230nm;
Column temperature: 25 DEG C;
Volumetric flow rate: 0.8mlmin
-1;
Sample size: 15 μ l;
With high performance liquid chromatograph, sample solution is detected, according to the content of the peak area recorded by 3,3'-dimethylellagic acids in linear equation calculation sample and 3,3', 4'-trimethyl ellagic acid.
The assay of 2 batch sample the results are shown in Table 2.
embodiment 13-14
The quantitative detecting method of 3,3'-dimethylellagic acids, 3,3', 4'-trimethyl ellagic acids in runcinate knotweed herb medicinal material, wherein, runcinate knotweed herb medicinal material is respectively 2 batches that gather spring, comprises the following steps:
(1) sample solution preparation: get runcinate knotweed herb medicinal material, pulverizes, and crosses 60 mesh sieves, take 1.0g, accurately weighed, be placed in container, add 100ml alcohol reflux and extract 1h, reclaim the solvent in extract and be concentrated into medicinal extract shape, adding 20ml water, ultrasonic process 20min, extracts 5 times with ethyl acetate jolting, and the consumption of ethyl acetate is each 20ml, evaporate to dryness acetic acid ethyl acetate extract, residue adds methyl alcohol and dissolves and be settled in 10ml measuring bottle, shakes up, 0.45 μm of miillpore filter filters, obtained sample solution;
(2) sample size measures:
Chromatographic condition:
C18 chromatographic column, enlightening horse Dikma ODS C18 post, the length of chromatographic column is 250mm, and diameter is 4.6mm, and packing material size is 5 μm;
Mobile phase: volume ratio is the methyl alcohol of 20: 30: 50: acetonitrile: 0.1% phosphoric acid;
Determined wavelength: 270nm;
Column temperature: 40 DEG C;
Volumetric flow rate: 1.2mlmin
-1;
Sample size: 20 μ l;
With high performance liquid chromatograph, sample solution is detected, according to the content of the peak area recorded by 3,3'-dimethylellagic acids in linear equation calculation sample and 3,3', 4'-trimethyl ellagic acid.
The assay of 2 batch sample the results are shown in Table 3.
According to the assay result of above 14 batch samples, this product should be plucked in the autumn in summer, and considers the fluctuation of medicinal material content, determine that the content of this product 3,3'-dimethylellagic acid must not be less than 0.5mg/g, 3, the content of 3', 4'-trimethyl ellagic acid must not be less than 0.17mg/g(in dry product).
below by test, the indexs such as precision of the present invention, stability and durability are measured.
1. precision test: precision measures the same standard solution of 10 μ L, continuous 6 sample introductions, measure by above-mentioned chromatographic condition, record chromatographic peak peak area.The RSD of 3,3'-dimethylellagic acid and 3,3', 4'-trimethyl ellagic acid, 6 sample introduction peak areas is respectively 0.61% and 1.64%, shows that instrument precision is good, in table 4.
4. average recovery test: 6 parts, the runcinate knotweed herb sample getting known content, every part of 0.5g, accurately weighed, precision adds a certain amount of 3,3'-dimethylellagic acids and 3,3', 4'-trimethyl ellagic acid reference substance, by need testing solution, preparation method prepares need testing solution, measures by above-mentioned chromatographic condition, calculate the recovery, measurement result is in table 7 and table 8.
5. the impact of wavelength variations: the same need testing solution of accurate absorption 10 μ L, under 246nm, 248nm, 250nm condition, difference sample introduction twice, measures according to above-mentioned chromatographic condition, record chromatographic peak peak area.The RSD of 3,3'-dimethylellagic acid and 3,3', 4'-trimethyl ellagic acid, 6 sample introduction peak areas is respectively 0.44% and 0.51%.Show that method measurement result when wavelength has less change has good stability, in table 10.
6. the impact of different brands chromatographic column: select Shimadzu, the separation case of C18 chromatographic column to runcinate knotweed herb medicinal material of Di Ma and Yi Li top grade different brands compare, find that three kinds of chromatographic columns all can be separated 3 preferably, 3'-dimethylellagic acid and 3,3', 4'-trimethyl ellagic acid.
From the result of above test, utilize the detection method of the present invention program, the indexs such as precision, stability and durability are all better.
The above, it is only preferred embodiment of the present invention, not any pro forma restriction is done to the present invention, anyly do not depart from technical solution of the present invention content, the any simple modification done above embodiment according to technical spirit of the present invention, equivalent variations and modification, all still belong in the scope of technical solution of the present invention.
Claims (9)
1. the quantitative detecting method of 3,3'-dimethylellagic acids, 3,3', 4'-trimethyl ellagic acids in a runcinate knotweed herb, comprises the following steps:
(1) sample solution preparation: get runcinate knotweed herb medicinal material, pulverizes, and crosses 60 mesh sieves, take 1.0g, accurately weighed, be placed in container, add 100ml alcohol reflux and extract 1h, reclaim the solvent in extract and be concentrated into medicinal extract shape, adding 20ml water, ultrasonic process 20min, extracts 5 times with ethyl acetate jolting, and the consumption of ethyl acetate is each 20ml, evaporate to dryness acetic acid ethyl acetate extract, residue adds methyl alcohol and dissolves and be settled in 10ml measuring bottle, shakes up, 0.45 μm of miillpore filter filters, obtained sample solution;
(2) sample size measures:
Chromatographic condition:
C18 chromatographic column;
Mobile phase: volume ratio is the methyl alcohol of 7-27: 20-50: 23-73: acetonitrile: 0.1% phosphoric acid;
Determined wavelength: 230-270nm;
Column temperature: 25-40 DEG C;
Volumetric flow rate: 0.8-1.2mlmin
-1;
Sample size: 10-20 μ l;
With high performance liquid chromatograph, sample solution is detected, according to the content of the peak area recorded by 3,3'-dimethylellagic acids in linear equation calculation sample and 3,3', 4'-trimethyl ellagic acid:
The linear equation of 3,3'-dimethylellagic acid is: Y
1=5598X
1+ 180.8, r=0.9991;
The linear equation of 3,3', 4'-trimethyl ellagic acid is: Y
2=5517.4X
2+ 73.3, r=0.9990;
Wherein, Y
1represent the peak area that 3,3'-dimethylellagic acid utilizes high performance liquid chromatograph to detect, Y
2represent the peak area that 3,3', 4'-trimethyl ellagic acid utilizes high performance liquid chromatograph to detect, X
1represent the sample introduction quality of 3,3'-dimethylellagic acid, X
2represent the sample introduction quality of 3,3', 4'-trimethyl ellagic acid, r is related coefficient.
2. 3,3'-dimethylellagic acids, 3,3' in a kind of runcinate knotweed herb as claimed in claim 1, the quantitative detecting method of 4'-trimethyl ellagic acid, wherein: described 3,3'-dimethylellagic acids, 3, the linear equation of 3', 4'-trimethyl ellagic acid obtains by the following method:
Get 3,3'-dimethylellagic acid reference substance 19.8mg and 3,3', 4'-trimethyl ellagic acid reference substance 9.82mg, adds methyl alcohol and dissolves and constant volume, makes every 1ml containing 3, the mixing reference substance stock solution of 3'-dimethylellagic acid 0.0990mg and 3,3', 4'-trimethyl ellagic acid 0.0491mg;
Precision measure mixing reference substance stock solution 2,4,6,8mL, put in 10mL measuring bottle, add methanol dilution constant volume, be mixed with gradient solution, make control series product solution together with former mixing reference substance stock solution;
Adopt chromatographic condition: C18 chromatographic column, mobile phase: volume ratio is the methyl alcohol of 7-27: 20-50: 23-73: acetonitrile: 0.1% phosphoric acid, determined wavelength: 230-270nm, column temperature: 25-40 DEG C, volumetric flow rate: 0.8-1.2mlmin
-1, sample size: 10-20 μ l, utilizes high performance liquid chromatograph to detect, with the sample introduction quality of reference substance be horizontal ordinate, peak area for ordinate, will sample introduction quality and the peak area of above-mentioned variable concentrations titer be recorded, by linear regression and get final product.
3. 3,3'-dimethylellagic acids, 3,3' in a kind of runcinate knotweed herb as claimed in claim 1 or 2, the quantitative detecting method of 4'-trimethyl ellagic acid, wherein: described C18 chromatographic column is Dikma ODS C18 post, and the length of chromatographic column is 250mm, diameter is 4.6mm, and packing material size is 5 μm.
4. the quantitative detecting method of 3,3'-dimethylellagic acids, 3,3', 4'-trimethyl ellagic acids in a kind of runcinate knotweed herb as claimed in claim 1 or 2, wherein: the first of described mobile phase to be volume ratio be 17:33:50: acetonitrile: 0.1% phosphoric acid.
5. in a kind of runcinate knotweed herb as claimed in claim 1 or 23, the quantitative detecting method of 3'-dimethylellagic acid, 3,3', 4'-trimethyl ellagic acids, wherein: the methyl alcohol in described mobile phase and acetonitrile are Chromatographic Pure Methanol and acetonitrile, described phosphoric acid is for analyzing pure phosphoric acid.
6. the quantitative detecting method of 3,3'-dimethylellagic acids, 3,3', 4'-trimethyl ellagic acids in a kind of runcinate knotweed herb as claimed in claim 1 or 2, wherein: described determined wavelength is 248nm.
7. the quantitative detecting method of 3,3'-dimethylellagic acids, 3,3', 4'-trimethyl ellagic acids in a kind of runcinate knotweed herb as claimed in claim 1 or 2, wherein: described column temperature is 30 DEG C.
8. the quantitative detecting method of 3,3'-dimethylellagic acids, 3,3', 4'-trimethyl ellagic acids in a kind of runcinate knotweed herb as claimed in claim 1 or 2, wherein: described volumetric flow rate is 1.0mlmin
-1.
9. the quantitative detecting method of 3,3'-dimethylellagic acids, 3,3', 4'-trimethyl ellagic acids in a kind of runcinate knotweed herb as claimed in claim 1 or 2, wherein: described sample size is 10 μ l.
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| CN106841464A (en) * | 2017-02-06 | 2017-06-13 | 广西壮族自治区中医药研究院 | Ellagic acid and 3,3` dimethoxy ellagic acid detection method of content in a kind of China fir parasitism |
| CN109765320A (en) * | 2019-03-12 | 2019-05-17 | 贵州远程制药有限责任公司 | A kind of content assaying method for spray agent for treating tendon and bone injury |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106841464A (en) * | 2017-02-06 | 2017-06-13 | 广西壮族自治区中医药研究院 | Ellagic acid and 3,3` dimethoxy ellagic acid detection method of content in a kind of China fir parasitism |
| CN109765320A (en) * | 2019-03-12 | 2019-05-17 | 贵州远程制药有限责任公司 | A kind of content assaying method for spray agent for treating tendon and bone injury |
| CN109765320B (en) * | 2019-03-12 | 2022-03-22 | 贵州远程制药有限责任公司 | Content determination method for tendon and bone injury spraying agent |
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