CN102495143A - Method for rapidly measuring content of three paeoniflorin compounds in white paeony root - Google Patents
Method for rapidly measuring content of three paeoniflorin compounds in white paeony root Download PDFInfo
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- CN102495143A CN102495143A CN2011103213082A CN201110321308A CN102495143A CN 102495143 A CN102495143 A CN 102495143A CN 2011103213082 A CN2011103213082 A CN 2011103213082A CN 201110321308 A CN201110321308 A CN 201110321308A CN 102495143 A CN102495143 A CN 102495143A
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Abstract
The invention discloses a method for rapidly measuring the content of three paeoniflorin compounds in white paeony root and belongs to the technical field of medicines. The method comprises the following steps of: (1) pre-processing a white paeony root sample to be measured; and (2) measuring the content of paeoniflorin sulfonate, paeoniflorin and albiflorin in the pre-processed white paeony root sample by high performance liquid chromatography or ultra-performance liquid chromatography. The measuring method is simple, convenient and rapid and has good accuracy and can be used for measuring the content of multiple components of the paeoniflorin compounds in the white paeony root.
Description
Technical field
The present invention relates to the method for three kinds of Paeoniflorin kind compound contents in the fast measuring root of herbaceous peony, belong to technical field of traditional Chinese medicines.
Background technology
The Chinese medicine root of herbaceous peony is the dry root of ranunculaceae plant Chinese herbaceous peony Paeonia Lactiflora pall., has the effect of nourishing blood for regulating menstruation, astringing YIN to stop sweating, easing the affected liver to relieve pain, suppressing liver-YANG.The Paeoniflorin compounds is the main active of generally acknowledging in the root of herbaceous peony, mainly is just to control (2010 editions " Chinese pharmacopoeia (an one), national medical sci-tech publishing houses: 96) according to paeoniflorin content to the quality of the root of herbaceous peony at present.
In the use of the root of herbaceous peony; Sulfur fumigation is as the maintenance process of a kind of traditional Chinese medicines material; Its objective is in order to let medicine materical crude slice rate of drying more attractive in appearance, Vermins-proof mildew-proof, quickening medicine materical crude slice in storage etc.; Have drying, brighten, insect protected, anticorrosion and anti-effect such as go mouldy, in the processing and storage process of Chinese crude drug and medicine materical crude slice, use general.Though sulfur fumigation has played certain positive role to the processing and storage of Chinese crude drug and medicine materical crude slice; But the proterties of Chinese crude drug and medicine materical crude slice changes more behind the modern study proof sulfur fumigation; And can make heavy metals such as wherein residual a large amount of sulphuric dioxide and arsenic, mercury, human body is had multiple harm; Because sulphuric dioxide is a kind of strong reductant, maybe with Chinese crude drug in contain the composition generation chemical reaction of ketone group, hydroxyl, cause loss of active ingredients; Therefore possibly change the nature and flavor of medicine, reduce the curative effect of medicine, thereby the quality that directly influences Chinese crude drug and medicine materical crude slice is (referring to Cheng Ming etc.; [J] inquired in the application that sulphur is smoked in Chinese crude drug processing and storage; China's traditional Chinese medicine information magazine, 2010,17 (5): 18).State Food and Drug Administration once sent the documents in 2004 and pointed out that the Chinese crude drug of sulfur fumigation was a medicinal material inferior." Chinese pharmacopoeia 2005 version (an one) has also been cancelled the stove drying technology of Chinese crude drugs such as Chinese yam, the root of kudzu vine, and in that " Chinese pharmacopoeia increased the detection of sulfur dioxide residual quantity in 2008 in the version enlarged edition.No longer advise using the method for sulfur fumigation at present for the processing and storage of Chinese crude drug and medicine materical crude slice.
But sulfur fumigation is still using in the process of manufacture of white Peony Root and medicine materical crude slice, and particularly serious in recent years, the white Peony Root and the medicine materical crude slice that circulate on the market are smoked through sulphur mostly.Because the difference of the place of production, job operation, wherein content of paeoniflorin difference is very big, and produces new chemical constitution such as Paeoniflorin sulfite; (referring to Wang Qiao etc.; The influence [J] to root of herbaceous peony chemical constitution, CHINA JOURNAL OF CHINESE MATERIA MEDICA, 2006 are concocted in processing; 31 (17): 1418), had a strong impact on the quality of white Peony Root and medicine materical crude slice.
China comprises in the root of herbaceous peony and measures existing bibliographical information by 8 component concentrations of above-mentioned three compounds (referring to Wang Qiao etc., the influence [J] to root of herbaceous peony chemical constitution, CHINA JOURNAL OF CHINESE MATERIA MEDICA are concocted in processing; 2006,31 (17): 1418), but these methods length all consuming time; Cost is high, is not suitable for conventional analysis.Still needleless is to three kinds of main Paeoniflorin compounds in the root of herbaceous peony at present, i.e. Paeoniflorin sulfite, Paeoniflorin and albiflorin carry out the method for quick assay.
Summary of the invention
In order to overcome the deficiency of prior art, the present invention provides the method for three kinds of Paeoniflorin kind compound contents in a kind of fast measuring root of herbaceous peony.
The present invention realizes through following technical scheme:
The method of three kinds of Paeoniflorin kind compound contents is characterized in that comprising the steps: in a kind of fast measuring root of herbaceous peony
(1) testing sample is carried out pre-service;
(2) pass through the content that high performance liquid chromatography or Ultra Performance Liquid Chromatography method are measured Paeoniflorin sulfite, Paeoniflorin and albiflorin in pretreated sample.
The pretreatment in the said step (1) is to extract 30-45 minute with 80% methyl alcohol ultrasonic (ultrasound condition: power 240W, frequency 45Khz).
The pretreatment method in the said step (1): precision takes by weighing the testing sample powder, accurate 80% methyl alcohol, the ultrasonic Extraction 30 minutes (power 240W, frequency 45Khz) of adding; Put coldly, claim again to decide weight, supply the weight that subtracts mistake with isoconcentration methyl alcohol; Shake up, filter, get subsequent filtrate.
Above-mentioned pretreated testing sample is white Peony Root and the root of herbaceous peony medicine materical crude slice that comprises sulfur fumigation.
80% methyl alcohol is mass concentration.
The chromatographic condition of high performance liquid chromatography or Ultra Performance Liquid Chromatography method is in the said step (2): adopt the C18 post, and ultraviolet detection, moving phase is by A phase and B phase composition, and A is 100% acetonitrile mutually, and B is phosphate aqueous solution mutually, adopts isocratic elution.
The particle diameter of said C18 post is 1.7～5.0 μ m, and column internal diameter 2.1～4.6mm, column length are 5～25cm; The detection wavelength of said UV-detector is 230nm; The concentration of B phase is 0.02%～0.03%; The A phase: the B phase volume ratio is 13%～15%: 85%～87%, and flow velocity is 0.3～1.0ml/min.
Concentration the best of B phase is 0.02%, and volume ratio most preferably is 85%.
The C18 post is a SHISEIDO CAPCELL PAK C18 post, or Waters ACQUITY UPLC BEH C18 post.
Advantage of the present invention is:
(1) the inventive method is for be applied to white Peony Root, detect when sulphur is smoked Paeoniflorin sulfite in the root of herbaceous peony, Paeoniflorin and albiflorin and analyze first.Paeoniflorin and albiflorin are the main effective constituent in the root of herbaceous peony, and the Paeoniflorin sulfite is to smoke the noval chemical compound that process produces at sulphur.This method has great importance to the detection of Paeoniflorin constituents in the root of herbaceous peony, and gordian techniquies such as new drug are had good directive significance, can be applicable to many aspects such as new drug development Quality Control.
(2) the inventive method is easy fast, accuracy is good; Through high performance liquid chromatography or Ultra Performance Liquid Chromatography method root of herbaceous peony sample is carried out the sample introduction analysis; Measure the content of three kinds of Paeoniflorin compounds in the root of herbaceous peony, analysis cost is low, the time is short, and stronger practicality is arranged.
Description of drawings
Fig. 1 is the high-efficient liquid phase chromatogram of Paeoniflorin sulfite, albiflorin and Paeoniflorin hybrid standard article solution.
Wherein chromatographic peak 1 is the Paeoniflorin sulfite, and chromatographic peak 2 is an albiflorin, and chromatographic peak 3 is a Paeoniflorin.
Fig. 2 smokes the high-efficient liquid phase chromatogram of root of herbaceous peony sample solution for sulphur.
Fig. 3 is the high-efficient liquid phase chromatogram of root of herbaceous peony sample solution.
Fig. 4 is the Ultra Performance Liquid Chromatography figure of Paeoniflorin sulfite, albiflorin and Paeoniflorin hybrid standard article solution.
Wherein chromatographic peak 1 is the Paeoniflorin sulfite, and chromatographic peak 2 is an albiflorin, and chromatographic peak 3 is a Paeoniflorin.
Fig. 5 smokes the Ultra Performance Liquid Chromatography figure of root of herbaceous peony sample solution for sulphur.
Fig. 6 is the Ultra Performance Liquid Chromatography figure of root of herbaceous peony sample solution.
Embodiment
Sulphur is smoked the high performance liquid chromatography assay of Paeoniflorin sulfite, albiflorin and Paeoniflorin in the root of herbaceous peony
(1) preparation of need testing solution: get sulphur and smoke the about 0.2g of White Peony Root (No. three sieves), the accurate title, decide, and puts in the tool plug conical flask, the accurate 80% methyl alcohol 20ml that adds; Claim to decide weight, sonicated (power 240W, frequency 45Khz) 30 minutes is put cold; Claim again to decide weight, supply the weight that subtracts mistake, shake up with 80% methyl alcohol; Filter, get subsequent filtrate, promptly get need testing solution.
(2) high-efficient liquid phase chromatogram condition: SHISEIDO CAPCELL PAK C18 chromatographic column (250mm * 4.6mm, 5 μ m); Moving phase: acetonitrile-0.02% phosphoric acid (15: 85); Elution time 20 minutes; Ultraviolet detection wavelength 230nm; Flow velocity 1.0ml.min-1; 30 ℃ of column temperatures; Sample size 10 μ l.
(3) preparation of reference substance solution: it is an amount of to get albiflorin reference substance, Paeoniflorin, Paeoniflorin sulfite reference substance, accurately claims surely, adds methyl alcohol and processes the solution that every 1ml contains 80 μ g, 160 μ g, 80 μ g respectively, promptly gets reference substance solution.
(4) linear relationship is investigated: it is an amount of to get albiflorin, Paeoniflorin, Paeoniflorin sulfite reference substance; The accurate title, decide; Add dissolve with methanol and be diluted to scale; Shake up, process and contain Paeoniflorin sulfite 2.09mg/ml, albiflorin 1.99mg/ml, Paeoniflorin 2.35mg/ml reference substance stock solution.The accurate respectively reference substance stock solution of drawing different volumes is processed the series of variable concentrations and is mixed reference substance solution.Measuring by above-mentioned HPLC chromatographic condition respectively, is abscissa with the concentration of reference substance, and the peak area integrated value is an ordinate, carries out regression treatment, calculates typical curve respectively.
(5) high effective liquid chromatography for measuring of root of herbaceous peony need testing solution: root of herbaceous peony need testing solution is carried out high effective liquid chromatography for measuring; Obtain the peak area of Paeoniflorin sulfite in the root of herbaceous peony need testing solution, albiflorin and Paeoniflorin chromatographic peak respectively; Equation of linear regression according to Paeoniflorin sulfite, albiflorin and Paeoniflorin; Calculate that Paeoniflorin sulfite, albiflorin and content of paeoniflorin are respectively 1.478%, 0.799% and 1.831% in the root of herbaceous peony, accomplish the assay that sulphur is smoked root of herbaceous peony sample.See Fig. 1 and Fig. 2.
Sulphur is smoked the Ultra Performance Liquid Chromatography method assay of Paeoniflorin sulfite, albiflorin and Paeoniflorin in the root of herbaceous peony
(1) preparation of need testing solution: get sulphur and smoke the about 0.2g of White Peony Root (No. three sieves), the accurate title, decide, and puts in the tool plug conical flask, and the accurate 80% methyl alcohol 20ml that adds claims to decide weight; Sonicated (power 240W, frequency 45Khz) 30 minutes is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with 80% methyl alcohol; Shake up, the accurate 5ml that draws to the 20ml measuring bottle, adds 80% methyl alcohol to scale; Shake up, filter, get subsequent filtrate, promptly get need testing solution.
(2) high-efficient liquid phase chromatogram condition: Waters ACQUITY UPLC BEH C18 chromatographic column (50mm * 2.1mm, 1.7 μ m); Moving phase: acetonitrile-0.02% phosphoric acid solution (13: 87); Elution time 3.5 minutes; Detect wavelength 230nm; Flow velocity 0.3mlmin-1; 30 ℃ of column temperatures; Sample size 2 μ l.
(3) preparation of reference substance solution: it is an amount of to get albiflorin reference substance, Paeoniflorin, Paeoniflorin sulfite reference substance, accurately claims surely, adds methyl alcohol and processes the solution that every 1ml contains 20 μ g, 40 μ g, 20 μ g respectively, promptly gets reference substance solution.
(4) linear relationship is investigated: it is an amount of to get albiflorin, Paeoniflorin, Paeoniflorin sulfite reference substance; The accurate title, decide; Add dissolve with methanol and be diluted to scale; Shake up, process and contain Paeoniflorin sulfite 2.09mg/ml, albiflorin 1.99mg/ml, Paeoniflorin 2.35mg/ml reference substance stock solution.The accurate respectively reference substance stock solution of drawing different volumes is processed the series of variable concentrations and is mixed reference substance solution.Measuring by above-mentioned HPLC chromatographic condition respectively, is abscissa with the concentration of reference substance, and the peak area integrated value is an ordinate, carries out regression treatment, calculates typical curve respectively.
(5) the Ultra Performance Liquid Chromatography method of root of herbaceous peony need testing solution is measured: root of herbaceous peony need testing solution is carried out the Ultra Performance Liquid Chromatography method measure; Obtain the peak area of Paeoniflorin sulfite in the root of herbaceous peony need testing solution, albiflorin and Paeoniflorin chromatographic peak respectively; Equation of linear regression according to Paeoniflorin sulfite, albiflorin and Paeoniflorin; Calculate that Paeoniflorin sulfite, albiflorin and content of paeoniflorin are respectively 1.474%, 0.793% and 1.824% in the root of herbaceous peony, accomplish the assay that sulphur is smoked root of herbaceous peony sample.(seeing Fig. 4 and Fig. 5)
The high performance liquid chromatography assay of Paeoniflorin sulfite, albiflorin and Paeoniflorin in the root of herbaceous peony
(1) preparation of need testing solution: get the about 0.2g of white Peony Root powder (No. three sieves), the accurate title, decide, and puts in the tool plug conical flask, the accurate 80% methyl alcohol 20ml that adds; Claim to decide weight, sonicated (power 240W, frequency 45Khz) 30 minutes is put cold; Claim again to decide weight, supply the weight that subtracts mistake, shake up with 80% methyl alcohol; Filter, get subsequent filtrate, promptly get need testing solution.
(2) high-efficient liquid phase chromatogram condition: SHISEIDO CAPCELL PAK C18 chromatographic column (250mm * 4.6mm, 5 μ m); Moving phase: acetonitrile-0.02% phosphoric acid (15: 85); Elution time 20 minutes; Detect wavelength 230nm; Flow velocity 1.0ml.min-1; 30 ℃ of column temperatures; Sample size 10 μ l.
(3) preparation of reference substance solution: it is an amount of to get albiflorin reference substance, Paeoniflorin, Paeoniflorin sulfite reference substance, accurately claims surely, adds methyl alcohol and processes the solution that every 1ml contains 80 μ g, 160 μ g, 80 μ g respectively, promptly gets reference substance solution.
(4) linear relationship is investigated: it is an amount of to get albiflorin, Paeoniflorin, Paeoniflorin sulfite reference substance; The accurate title, decide; Add dissolve with methanol and be diluted to scale; Shake up, process and contain Paeoniflorin sulfite 2.09mg/ml, albiflorin 1.99mg/ml, Paeoniflorin 2.35mg/ml reference substance stock solution.The accurate respectively reference substance stock solution of drawing different volumes is processed the series of variable concentrations and is mixed reference substance solution.Measuring by above-mentioned HPLC chromatographic condition respectively, is abscissa with the concentration of reference substance, and the peak area integrated value is an ordinate, carries out regression treatment, calculates typical curve respectively.
(5) high effective liquid chromatography for measuring of root of herbaceous peony need testing solution: root of herbaceous peony need testing solution is carried out high effective liquid chromatography for measuring; Obtain the peak area of Paeoniflorin sulfite in the root of herbaceous peony need testing solution, albiflorin and Paeoniflorin chromatographic peak respectively; Equation of linear regression according to Paeoniflorin sulfite, albiflorin and Paeoniflorin; Calculate that albiflorin and content of paeoniflorin are respectively 0.665% and 3.258% in the root of herbaceous peony; Do not detect the Paeoniflorin sulfite, accomplish the assay of white Peony Root sample.(seeing Fig. 1 and Fig. 3)
Embodiment 4
The Ultra Performance Liquid Chromatography method assay of Paeoniflorin sulfite, albiflorin and Paeoniflorin in the root of herbaceous peony
(1) preparation of need testing solution: get the about 0.2g of white Peony Root powder (No. three sieves), the accurate title, decide, and puts in the tool plug conical flask, and the accurate 80% methyl alcohol 20ml that adds claims to decide weight; Sonicated (power 240W, frequency 45Khz) 30 minutes is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with 80% methyl alcohol; Shake up, the accurate 5ml that draws to the 20ml measuring bottle, adds 80% methyl alcohol to scale; Shake up, filter, get subsequent filtrate, promptly get need testing solution.
(2) high-efficient liquid phase chromatogram condition: Waters ACQUITY UPLC BEH C18 chromatographic column (50mm * 2.1mm, 1.7 μ m); Moving phase: acetonitrile-0.02% phosphoric acid solution (13: 87); Elution time 3.5 minutes; Detect wavelength 230nm; Flow velocity 0.3mlmin-1; 30 ℃ of column temperatures; Sample size 2 μ l.
(3) preparation of reference substance solution: it is an amount of to get albiflorin reference substance, Paeoniflorin, Paeoniflorin sulfite reference substance, accurately claims surely, adds methyl alcohol and processes the solution that every 1ml contains 20 μ g, 40 μ g, 20 μ g respectively, promptly gets reference substance solution.
(4) linear relationship is investigated: it is an amount of to get albiflorin, Paeoniflorin, Paeoniflorin sulfite reference substance; The accurate title, decide; Add dissolve with methanol and be diluted to scale; Shake up, process and contain Paeoniflorin sulfite 2.09mg/ml, albiflorin 1.99mg/ml, Paeoniflorin 2.35mg/ml reference substance stock solution.The accurate respectively reference substance stock solution of drawing different volumes is processed the series of variable concentrations and is mixed reference substance solution.Measuring by above-mentioned HPLC chromatographic condition respectively, is abscissa with the concentration of reference substance, and the peak area integrated value is an ordinate, carries out regression treatment, calculates typical curve respectively.
(5) the Ultra Performance Liquid Chromatography method of root of herbaceous peony need testing solution is measured: root of herbaceous peony need testing solution is carried out the Ultra Performance Liquid Chromatography method measure; Obtain the peak area of Paeoniflorin sulfite in the root of herbaceous peony need testing solution, albiflorin and Paeoniflorin chromatographic peak respectively; Equation of linear regression according to Paeoniflorin sulfite, albiflorin and Paeoniflorin; Calculate that albiflorin and content of paeoniflorin are respectively 0.660% and 3.252% in the root of herbaceous peony; Do not detect the Paeoniflorin sulfite, accomplish the assay of white Peony Root sample.(seeing Fig. 4 and Fig. 6)
Embodiment 5
Sulphur is smoked the high performance liquid chromatography assay of Paeoniflorin sulfite, albiflorin and Paeoniflorin in the root of herbaceous peony
(1) preparation of need testing solution: get sulphur and smoke the about 0.2g of White Peony Root (No. three sieves), the accurate title, decide, and puts in the tool plug conical flask, the accurate 80% methyl alcohol 20ml that adds; Claim to decide weight, sonicated (power 240W, frequency 45Khz) 45 minutes is put cold; Claim again to decide weight, supply the weight that subtracts mistake, shake up with 80% methyl alcohol; Filter, get subsequent filtrate, promptly get need testing solution.
(2) high-efficient liquid phase chromatogram condition: SHISEIDO CAPCELL PAK C18 chromatographic column (250mm * 4.6mm, 5 μ m); Moving phase: acetonitrile-0.02% phosphoric acid (15: 85); Elution time 20 minutes; Ultraviolet detection wavelength 230nm; Flow velocity 0.8ml.min-1; 30 ℃ of column temperatures; Sample size 10 μ l.
(3) preparation of reference substance solution: it is an amount of to get albiflorin reference substance, Paeoniflorin, Paeoniflorin sulfite reference substance, accurately claims surely, adds methyl alcohol and processes the solution that every 1ml contains 80 μ g, 160 μ g, 80 μ g respectively, promptly gets reference substance solution.
(4) linear relationship is investigated: it is an amount of to get albiflorin, Paeoniflorin, Paeoniflorin sulfite reference substance; The accurate title, decide; Add dissolve with methanol and be diluted to scale; Shake up, process and contain Paeoniflorin sulfite 2.09mg/ml, albiflorin 1.99mg/ml, Paeoniflorin 2.35mg/ml reference substance stock solution.The accurate respectively reference substance stock solution of drawing different volumes is processed the series of variable concentrations and is mixed reference substance solution.Measuring by above-mentioned HPLC chromatographic condition respectively, is abscissa with the concentration of reference substance, and the peak area integrated value is an ordinate, carries out regression treatment, calculates typical curve respectively.
(5) high effective liquid chromatography for measuring of root of herbaceous peony need testing solution: root of herbaceous peony need testing solution is carried out high effective liquid chromatography for measuring; Obtain the peak area of Paeoniflorin sulfite in the root of herbaceous peony need testing solution, albiflorin and Paeoniflorin chromatographic peak respectively; Equation of linear regression according to Paeoniflorin sulfite, albiflorin and Paeoniflorin; Calculate that Paeoniflorin sulfite, albiflorin and content of paeoniflorin are respectively 1.580%, 0.541% and 1.218% in the root of herbaceous peony, accomplish the assay that sulphur is smoked root of herbaceous peony sample.
Embodiment 6
Sulphur is smoked the high performance liquid chromatography assay of Paeoniflorin sulfite, albiflorin and Paeoniflorin in the root of herbaceous peony
(1) preparation of need testing solution: get sulphur and smoke the about 0.2g of White Peony Root (No. three sieves), the accurate title, decide, and puts in the tool plug conical flask, the accurate 80% methyl alcohol 20ml that adds; Claim to decide weight, sonicated (power 240W, frequency 45Khz) 30 minutes is put cold; Claim again to decide weight, supply the weight that subtracts mistake, shake up with 80% methyl alcohol; Filter, get subsequent filtrate, promptly get need testing solution.
(2) high-efficient liquid phase chromatogram condition: SHISEIDO CAPCELL PAK C18 chromatographic column (250mm * 4.6mm, 5 μ m); Moving phase: acetonitrile-0.03% phosphoric acid (14: 86); Elution time 20 minutes; Ultraviolet detection wavelength 230nm; Flow velocity 1.0ml.min-1; 30 ℃ of column temperatures; Sample size 10 μ l.
(3) preparation of reference substance solution: it is an amount of to get albiflorin reference substance, Paeoniflorin, Paeoniflorin sulfite reference substance, accurately claims surely, adds methyl alcohol and processes the solution that every 1ml contains 80 μ g, 160 μ g, 80 μ g respectively, promptly gets reference substance solution.
(4) linear relationship is investigated: it is an amount of to get albiflorin, Paeoniflorin, Paeoniflorin sulfite reference substance; The accurate title, decide; Add dissolve with methanol and be diluted to scale; Shake up, process and contain Paeoniflorin sulfite 2.09mg/ml, albiflorin 1.99mg/ml, Paeoniflorin 2.35mg/ml reference substance stock solution.The accurate respectively reference substance stock solution of drawing different volumes is processed the series of variable concentrations and is mixed reference substance solution.Measuring by above-mentioned HPLC chromatographic condition respectively, is abscissa with the concentration of reference substance, and the peak area integrated value is an ordinate, carries out regression treatment, calculates typical curve respectively.
(5) high effective liquid chromatography for measuring of root of herbaceous peony need testing solution: root of herbaceous peony need testing solution is carried out high effective liquid chromatography for measuring; Obtain the peak area of Paeoniflorin sulfite in the root of herbaceous peony need testing solution, albiflorin and Paeoniflorin chromatographic peak respectively; Equation of linear regression according to Paeoniflorin sulfite, albiflorin and Paeoniflorin; Calculate that Paeoniflorin sulfite, albiflorin and content of paeoniflorin are respectively 3.322%, 0.676% and 0.585% in the root of herbaceous peony, accomplish the assay that sulphur is smoked root of herbaceous peony sample.
Claims (8)
1. the method for three kinds of Paeoniflorin kind compound contents in the fast measuring root of herbaceous peony is characterized in that comprising the steps:
(1) testing sample is carried out pre-service;
(2) pass through the content that high performance liquid chromatography or Ultra Performance Liquid Chromatography method are measured Paeoniflorin sulfite, Paeoniflorin and albiflorin in pretreated sample;
2. method according to claim 1 is characterized in that, The pretreatment is ultrasonic with 80% methyl alcohol in the step (1), extracts ultrasound condition: power 240W, frequency 45Khz 30-45 minute.
3. method according to claim 2 is characterized in that, The pretreatment method in the step (1): precision takes by weighing the testing sample powder, accurate 80% methyl alcohol that adds; Ultrasonic Extraction 30 minutes is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with isoconcentration methyl alcohol; Shake up, filter, get subsequent filtrate; Ultrasound condition: power 240W, frequency 45Khz.
4. according to claim 1 or 2 or 3 described methods, it is characterized in that pretreated testing sample is white Peony Root and the root of herbaceous peony medicine materical crude slice that comprises sulfur fumigation.
5. method according to claim 1 is characterized in that the chromatographic condition of middle high performance liquid chromatography of step (2) or Ultra Performance Liquid Chromatography method is: adopt the C18 post, ultraviolet detection; Moving phase is by A phase and B phase composition; A is 100% acetonitrile mutually, and B is phosphate aqueous solution mutually, adopts isocratic elution.
6. method according to claim 5 is characterized in that, the particle diameter of said C18 post is 1.7～5.0 μ m, and column internal diameter 2.1～4.6mm, column length are 5～25cm; Said ultraviolet detection wavelength is 230nm; The concentration of B phase is 0.02%～0.03%; The A phase: the B phase volume ratio is 13%～15%: 85%～87%, and flow velocity is 0.3～1.0ml/min.
7. method according to claim 6, the concentration that it is characterized in that the B phase is 0.02%, volume ratio 85%.
8. according to claim 5 or 6 described methods, it is characterized in that said C18 post is a SHISEIDO CAPCELL PAK C18 post, or Waters ACQUITY UPLC BEH C18 post.
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| CN105116086A (en) * | 2015-08-18 | 2015-12-02 | 辽宁大学 | HPLC-ELSD content determination method for paeoniflorin and albiflori in red paeony roots |
| CN108459111A (en) * | 2018-05-09 | 2018-08-28 | 国珍健康科技（北京）有限公司 | A method of paeoniflorin content is measured by high performance liquid chromatography |
| CN111650304A (en) * | 2020-06-29 | 2020-09-11 | 山西大学 | Method for rapidly screening sulphur-smoked flos farfarae |
| CN114740115A (en) * | 2022-04-19 | 2022-07-12 | 青岛市食品药品检验研究院（青岛市药品不良反应监测中心、青岛市实验动物和动物实验中心） | Radix paeoniae alba fumigation detection method |
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| CN102998386A (en) * | 2012-11-22 | 2013-03-27 | 宁波立华制药有限公司 | Method for measuring radix paeoniae alba diglucoside in radix paeoniae alba callus by utilizing high performance liquid chromatography |
| CN105116086A (en) * | 2015-08-18 | 2015-12-02 | 辽宁大学 | HPLC-ELSD content determination method for paeoniflorin and albiflori in red paeony roots |
| CN108459111A (en) * | 2018-05-09 | 2018-08-28 | 国珍健康科技（北京）有限公司 | A method of paeoniflorin content is measured by high performance liquid chromatography |
| CN111650304A (en) * | 2020-06-29 | 2020-09-11 | 山西大学 | Method for rapidly screening sulphur-smoked flos farfarae |
| CN114740115A (en) * | 2022-04-19 | 2022-07-12 | 青岛市食品药品检验研究院（青岛市药品不良反应监测中心、青岛市实验动物和动物实验中心） | Radix paeoniae alba fumigation detection method |
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