CN101354381A - Method for evaluating quality of loquat leaf medicinal materials - Google Patents
Method for evaluating quality of loquat leaf medicinal materials Download PDFInfo
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Abstract
The invention relates to a method for building a fingerprint of loquat leaf medicinal materials and a fingerprint thereof. The method comprises the following steps of: firstly collecting the loquat leaf medicinal materials for over 10 batches, preparing test sample solution and a reference product solution, implementing measurement by referring to <Technical requirements of fingerprint research of traditional Chinese medicine injection (Provisional)> and applying a high-efficiency liquid chromatography method and a high-efficiency capillary electrophoresis method, and finally implementing analysis by applying a computation software of the similarity between the fingerprint of traditional Chinese medicines (which is recommended by the State Pharmacopoeia Committee). The method of the invention has the advantages of simplicity, stability, high precision, good reproducibility and easy grasp, can grasp the variety and the quality condition of the loquat leaves from the integral characteristic face and the high profile of chromatogram, and can be used as one of the methods for evaluating the quality of the loquat leaves and distinguishing the loquat leaves between true and false.
Description
Technical field
The present invention relates to the discriminating and the quality evaluating method of loguat leaf medicinal material, specifically differentiate and estimate the loguat leaf method for quality, relate in particular to high performance capillary electrophoresis finger-print or efficient liquid-phase chromatograph finger print atlas and differentiate the loguat leaf true and false and stay-in-grade method with finger-print.
Background technology
Loguat leaf is the dried leaves of rosaceous plant loquat Eriobotrya japonica (Thunb.) Lindl., it is large middle about material commonly used, has removing heat from the lung to relieve cough, effects such as stopping nausea and vomiting by lowering the adverse flow of QI, be used for the cough with lung heat, the circulation of vital energy in the wrong direction is breathed heavily urgency, and stomach energy is vomitted contrary, the thirsty grade of dysphoria with smothery sensation (Chinese Pharmacopoeia Commission compiles. Pharmacopoeia of People's Republic of China. and first one. Beijing: Chemical Industry Press, 2005:142).Discover (Lin Yulin in recent years in a large number, Lin Wenjin, it is strong to stand in great numbers. loguat leaf present Research and DEVELOPMENT PROSPECT, Chinese crude drug .2006,29 (10): 1111-1114), the main active of loguat leaf has ursolic acid, oleanolic acid, amygdalin, saponins, polysaccharide etc., have cough-relieving, eliminate the phlegm, anti-inflammatory, protect the liver, pain relieving, antitumor and regulate multiple pharmacological effect such as immunity; Clinically can be used for treating illnesss such as respiratory inflammation, rheumatic arthritis, glomerulonephritis, anaphylactoid purpura, nettle rash, eczema and hyperglycaemia.
The quality standard of relevant loguat leaf, " one one of Chinese pharmacopoeia is only checked simple proterties, physicochemical identification and water-soluble extractives to version in 2005, regulation is not made in the detection of effective constituent as yet.At present existing bibliographical information with the HPLC method measure the loguat leaf medicinal materials fingerprint research (Zhu Yanping is etc. loguat leaf high-efficiency liquid-phase fingerprint research for Zhang Lvhao, Wu Songji. the time precious traditional Chinese medical science traditional Chinese medicines .2007,18 (2): 269-271).But, do not appear in the newspapers at the efficient liquid-phase chromatograph finger print atlas of Fujian Province's authentic medicinal herbs loguat leaf because the mass discrepancy of zones of different loguat leaf is bigger.
Capillary Electrophoresis is a kind of high efficient separation technology, have efficient, fast, characteristics such as sampling volume is little, solvent consumption is few and contamination resistance is strong, demonstrating significant advantage (Ma Xin aspect Chinese medicine analysis of effective component, the finger-print research, Sun Yuqing. the capillary electrophoresis fingerprint research of ginkgo biloba p.e. Chinese herbal medicine, 2004,35 (8): 876-878).At present, do not see the report that adopts capillary electrophoresis fingerprint to differentiate and estimate the loguat leaf quality of medicinal material, the applicant has carried out investigating relatively to different places of production loguat leaf, builds up loguat leaf GAP demonstration plant base in 2002 in the Putian, Fujian, carries out the GAP management.For setting up the integrated evaluating method of loguat leaf quality of medicinal material, special with State Food and Drug Administration issued in 2000 about the requirement of traditional Chinese medicine fingerprint pattern technology, 10 batches of above loguat leaf medicinal materials to Fujian loquat main breed and loguat leaf GAP base are measured, and have set up Fujian authentic medicinal herbs loguat leaf finger print measuring method.
Summary of the invention
The purpose of this invention is to provide a kind of method, and can use it for one of index of loguat leaf quality control and real and fake discrimination thereof with finger-print discriminating and evaluation loguat leaf quality of medicinal material.
The present invention implements through the following steps:
The first step, the standard finger-print when setting up loquat earlier, key step comprises:
(a) gather representative loguat leaf medicinal material more than 10 batches, and identify its kind and quality.
(b) preparation of loguat leaf medicinal material standard solution: get the loguat leaf medicinal material, pulverize, sieve, be dried to constant weight.It is an amount of that precision takes by weighing Loquat Leaf, uses alcohol solvent (the C number is preferably methyl alcohol or normal butyl alcohol less than 5) ultrasonic Extraction 2～3 times, merges extract.Filter, filtrate adds alcohol solvent (the C number is preferably methyl alcohol or normal butyl alcohol less than 5) or moving phase and is diluted to finite concentration after concentrating, and shakes up, promptly.Filter with miillpore filter before the sample introduction.
The finger-print that adopts the high-performance capillary electrophoresis method to measure, the preferred step of concrete preparation of its loguat leaf medicinal material standard solution is: get the about 1g of Loquat Leaf (crossing sieve No. four), precision claims fixed, add 30 times of amount normal butyl alcohols, leave standstill 15min, ultrasonic Extraction 3 times, each 30min, filter, get filtrate, solvent is flung in heating, use dissolve with methanol, move to the 5ml measuring bottle, add methyl alcohol and be diluted to scale, shake up; Promptly.
Adopt the finger-print of high effective liquid chromatography for measuring, the concrete preparation process of its loguat leaf medicinal material standard solution is: get the loguat leaf medicinal material, pulverize, cross sieve No. four, powder is dried to constant weight for 60 ℃.Precision takes by weighing Loquat Leaf 1g, puts in the 50ml measuring bottle, uses methyl alcohol 30ml ultrasonic Extraction 2 times, and each 30min merges extract.Filter, filtrate concentrates in water-bath, moves in the 10ml measuring bottle, adds methyl alcohol and is diluted to scale, shakes up, promptly.
(c) preparation of object of reference solution: object of reference is the effective constituent in the loguat leaf, is preferably the triterpenic acid compounds, more preferably ursolic acid.The concrete preparation method of its test liquid is: precision takes by weighing ursolic acid reference substance 5mg and places the 10ml volumetric flask, with dissolve with methanol and be diluted to scale, shakes up, promptly.
(d) selection of assay method: assay method is a chromatography, is preferably high performance capillary electrophoresis (HPCE) or high performance liquid chromatography (HPLC).
(f) measure: accurate loguat leaf medicinal material standard solution and the object of reference solution drawn, inject detecting instrument respectively, measure the loguat leaf standard finger-print.
The finger-print that adopts the high-performance capillary electrophoresis method to measure, its preferred condition is: capillary column (60cm is long, 50 μ m internal diameters); Damping fluid: (0.1mol/L BAS: 0.2mol/L potassium hydroxide solution=4: 2); Separation voltage: 20kV; Experimental temperature: 25 ℃; The detection wavelength is 200nm.Number of theoretical plate calculates by the ursolic acid peak and is not less than 3000.
Adopt the finger-print of high effective liquid chromatography for measuring, its preferred condition is: chromatographic column: Eclipse XDB-C18 post (4.6mm * 150mm, 5 μ m), and moving phase: methyl alcohol: 1% aqueous acetic acid (90: 10), detect wavelength 215nm, flow velocity 1.0mlmin
-1, column temperature is a room temperature, sample size 10 μ l.Number of theoretical plate calculates by the ursolic acid peak and is not less than 2000.
In second step, measure loguat leaf sample finger-print to be measured with above-mentioned identical method;
In the 3rd step, with loguat leaf sample finger-print to be measured and the contrast of loguat leaf standard finger-print, screening meets the loguat leaf medicinal material of loguat leaf standard finger-print.
With the finger-print that the high-performance capillary electrophoresis method is measured, the relative standard deviation RSD of the relative retention time at its 9 total peaks is all less than 3%, and wherein the average relative retention time RT at No. 1 peak is 0.656, and relative standard deviation RSD is 0.62%; The average RT at No. 2 peaks is 0.712, and RSD is 0.50%; The average RT at No. 3 peaks is 0.818, and RSD is 1.08%; The average RT at No. 4 peaks is 0.871, and RSD is 0.34%; The average RT at No. 5 peaks is 0.892, and RSD is 0.30%; The average RT at No. 6 peaks is 0.919, and RSD is 0.29%; The average RT at No. 7 peaks is 0.953, and RSD is 0.62%; No. 8 peaks are ursolic acid, i.e. object of reference, and its average RT is 1.000, RSD is 0.00%; The average RT at No. 7 peaks is 1.070, and RSD is 0.54%.
With the finger-print of high effective liquid chromatography for measuring, the relative standard deviation RSD of the relative retention time at its 8 total peaks is all less than 3%, and wherein the average relative retention time RT at No. 1 peak is 0.322, and relative standard deviation RSD is 2.34%; The average RT at No. 2 peaks is 0.344, and RSD is 2.17%; The average RT at No. 3 peaks is 0.372, and RSD is 1.57%; The average RT at No. 4 peaks is 0.574, and RSD is 2.01%; The average RT at No. 5 peaks is 0.607, and RSD is 1.18%; The average RT at No. 6 peaks is 0.762, and RSD is 2.92%; The average RT at No. 7 peaks is 0.950, and RSD is 1.28%; No. 8 peaks are ursolic acid, i.e. object of reference, and its average RT is 1.000, RSD is 0.00%.
Advantage of the present invention is as follows:
(1) the loguat leaf finger-print of being set up can characterize the quality of loguat leaf medicinal material effectively, and the whole chemical constitutions in the loguat leaf are detected, and both unrealisticly also need not.A large amount of pharmacology and clinical research result show that the triterpenic acid constituents in the loguat leaf is one of its important effective constituent, so select for use ursolic acid wherein just very representative as the reference product.
(2) do as a wholely to treat with the whole finger-print of loguat leaf, pay attention to each front and back that constitute fingerprint characteristic peak order and mutual relationship, pay attention to whole facial feature, both avoided judging the one-sidedness of loguat leaf total quality because of only measuring one or two chemical constitution, reduced again and be the artificial possibility of handling of requisite quality, the present invention will contribute for quality and the curative effect that improves loguat leaf and Chinese patent medicine preparation thereof for quality complete, that accurately estimate loguat leaf provides new reference standard.
(3) the present invention have that method is easy, stable, precision is high, favorable reproducibility, the characteristics that are easy to grasp.The present invention adopts ultrasonic method to extract the preparation test liquid, cost and time that sample preparation is greatly easy, saved check.The present invention simultaneously proposes to adopt simultaneously the quality of the method for capillary electrophoresis fingerprint and high-efficiency liquid-phase fingerprint to same batch of medicinal material comprehensive evaluation loguat leaf medicinal material.
Description of drawings
Fig. 1 loguat leaf Capillary Electrophoresis chromatogram has peak (A is loguat leaf sample chromatogram figure, and B is a ursolic acid reference substance chromatogram)
Fig. 2 loguat leaf high-performance capillary electrophoresis stacking diagram
Fig. 3 loguat leaf high-efficient liquid phase chromatogram has peak (A is loguat leaf sample chromatogram figure, and B is a ursolic acid reference substance chromatogram)
Fig. 4 loguat leaf high performance liquid chromatography stacking diagram
Embodiment
The invention will be further described below in conjunction with embodiment, but content of the present invention is not limited to this fully.
The foundation of embodiment one loguat leaf high performance capillary electrophoresis fingerprint spectrum method
1 instrument and reagent
P/ACE MDQ capillary electrophoresis system (U.S. Beckman Coulter Inc.), diode array detector; Coating quartz capillary (Hebei light transmitting fiber Yongnian factory) not, METTLERAL204 electronic balance (Germany); KUDOS SK3310HP type Ultrasound Instrument (Shanghai); GZX-9070MBE constant temperature air dry oven (Shanghai); The Ai Kepu ultrapure water machine; Finger-print computer aided similarity evaluation system software (software is recommended by Chinese Pharmacopoeia Commission).Methyl alcohol is chromatographically pure, and normal butyl alcohol is pure for analyzing, and water is ultrapure water.The reference substance ursolic acid is available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number 110742-200415.
The loguat leaf test sample picks up from the extension tree Lao Ye in Putian City loguat leaf GAP medicinal material base, Fujian, sampling time is 01 day-Dec 20 Dec in 2005, through 60 ℃ of oven dry altogether 13 batches of samples, each source, the sample place of production and numbering see Table 1, the chamber identifies that each batch quality of medicinal material all meets the relevant regulations under loguat leaf item of Chinese Pharmacopoeia version in 2005 approximately in Fujian Province's medical science test key lab.
Table 1 experiment medicinal material complete list
2 methods and result
2.1 chromatographic condition and system suitability test: capillary column (60cm is long, 50 μ m internal diameters); Damping fluid: (0.1mol/L BAS: 0.2mol/L potassium hydroxide solution=4: 2); Separation voltage: 20kV; Experimental temperature: 25 ℃; The detection wavelength is 200nm.Number of theoretical plate calculates by the ursolic acid peak and is not less than 3000.
2.2 the preparation of object of reference solution: precision takes by weighing ursolic acid reference substance 5mg and places the 10ml volumetric flask, with dissolve with methanol and be diluted to scale, shakes up promptly.
2.3 the preparation of need testing solution: get the about 1g of this product powder (crossing sieve No. four), the accurate title, decide, and adds 30 times of amount normal butyl alcohols, leaves standstill 15min, ultrasonic Extraction 3 times, each 30min filters, and gets filtrate, and solvent is flung in heating, use dissolve with methanol, move to the 5ml measuring bottle, add methyl alcohol and be diluted to scale, shake up; 0.45 μ m miillpore filter filters, and is standby.
2.4 determination method: adopt the pressure sample introduction, sample size: 0.5psi * 10s, sample and reference substance are measured with method.
2.5 methodological study
2.5.1 precision test: get the ursolic acid reference substance solution, continuous sample introduction 5 times, the consistance of investigation chromatographic peak retention time and peak area, the retention time RSD of ursolic acid and peak area RSD are respectively 0.48% and 1.32%.
2.5.2 stability test: get the ursolic acid reference substance solution, get with a need testing solution, respectively 0,2,4,6,8, the 10h different time points is measured the consistance of investigating retention time and peak area, and the retention time RSD of ursolic acid and peak area RSD are respectively 1.57% and 1.84%.
2.5.3 reappearance test: get the ursolic acid reference substance solution, detect 5 parts with method, investigate the consistance of retention time and peak area, the retention time RSD of ursolic acid and peak area RSD are respectively 1.42% and 2.17%.
2.6 testing result: by the need testing solution of 13 batches of loguat leaf medicinal materials of above-mentioned 2.2.1 method preparation, sample detection successively under above-mentioned 2.1 selected same experiment conditions, the result records the HPLC chromatogram of 12 test samples.
2.7 finger-print similarity analysis
2.7.1 the selection at common characteristic peak according to 13 batches of given peak number, peak value (integrated value) and peak position correlation parameters such as (relative retention times) of test sample measurement result analyze, through contrasting with reference substance, and the chromatogram of all mensuration and record relatively, choose the characteristic peak of 9 total peaks as finger-print, wherein No. 8 peaks derive from ursolic acid and (see Fig. 1, A is loguat leaf sample chromatogram figure, and B is a ursolic acid reference substance chromatogram).
2.7.2 the foundation of total figure spectral model imports computer aided similarity evaluation system software with the test data of each batch sample, through selecting the peak, set matching template, will compose the peak and mate automatically, set up its total collection of illustrative plates standard form, carry out the similarity evaluation of each sample then.
2.7.3 the collection of illustrative plates similarity analysis adopts the similarity of each batch of computer aided similarity evaluation system software analysis sample finger-print, the results are shown in Figure 2, table 2～table 4.By analysis, the loguat leaf similarity of removing batch 10 (pinches) and batch 13 (the new bases of Pu) is below 0.9, and the loguat leaf sample HPCE fingerprint chromatogram similarity value of all the other 11 batches of different places of production and kind is all greater than 0.9.
Table 2 finger-print similarity analysis result
The relative retention time table of total fingerprint peaks in the table 3 loguat leaf kapillary finger-print
The relative area ratio of total fingerprint peaks in the table 4 loguat leaf kapillary finger-print
The foundation of embodiment two loguat leaf efficient liquid-phase chromatograph finger print atlas methods
1, instrument and reagent
The HP1100 high performance liquid chromatograph, Eclipse XDB-C18 post (4.6mm * 150mm, 5 μ m), diode array detector; METTLER AL204 electronic balance (Germany); KUDOS SK3310HP type Ultrasound Instrument (Shanghai); GZX-9070MBE constant temperature air dry oven (Shanghai); Finger-print computer aided similarity evaluation system software (Fujian Province's medical science test key lab writes).Methyl alcohol is chromatographically pure, and glacial acetic acid is pure for analyzing, and water is purified water.The reference substance ursolic acid is available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number 110742-200415.
The loguat leaf test sample picks up from the loguat leaf GAP medicinal material base, Chang Tai town (alpine region) of Fujian Putian City, Xin Du town fruit (region of no relief), four sampled points of Putian City fruit tree research institute (low-relief terrain) and Fujian Province's fruit tree research institute (low-relief terrain, Fuzhou City), sampling time is 01 day-October 20 October in 2005, gather the liberation clock respectively, early No. 6, clock, the extension tree Lao Ye of Changhong No. 3 (being Fujian Province's three big loquat main breeds), through 60 ℃ of oven dry altogether 12 batches of samples, each source, the sample place of production and numbering see Table 5, mesothecium identifies that each batch quality of medicinal material all meets the relevant regulations under loguat leaf item of Chinese Pharmacopoeia version in 2005 through Fujian Province's medical science test key lab.
Table 5 medicinal material sample source
2, method and result
2.1 chromatographic condition
Chromatographic column: Eclipse XDB-C18 post (4.6mm * 150mm, 5 μ m), moving phase: methyl alcohol: 1% aqueous acetic acid (90: 10), detect wavelength 215nm, flow velocity 1.0mlmin
-1, column temperature is a room temperature, sample size 10 μ l.Number of theoretical plate calculates by the ursolic acid peak and is not less than 2000.
2.2 the preparation of solution
2.2.1 the loguat leaf medicinal material is got in the preparation of need testing solution, pulverizes, and crosses sieve No. four, powder is dried to constant weight for 60 ℃.Precision takes by weighing Loquat Leaf 1g, puts in the 50ml measuring bottle, uses methyl alcohol 30ml ultrasonic Extraction 2 times, and each 30min merges extract.Filter, filtrate concentrates in water-bath, moves in the 10ml measuring bottle, adds methyl alcohol and is diluted to scale, shake up, and 0.45 μ m filtering with microporous membrane, promptly.
2.2.2 the preparation precision of reference substance solution takes by weighing ursolic acid reference substance 5mg and places the 10ml volumetric flask, with dissolve with methanol and be diluted to scale, shakes up promptly.
2.3 methodological study
2.3.1 the accurate reference substance solution 10 μ l that draw of precision test repeat sample introduction 6 times, the RSD of ursolic acid peak area is 1.20%, shows that precision is good.
2.3.2 stability test: get same need testing solution, 0,1,2,4,8,24h is sample introduction 10 μ l respectively, and the RSD of 6 sample introduction ursolic acid peak areas is 2.15% as a result, show that need testing solution is stable in 24h.
2.3.3 replica test: take by weighing 5 parts of same batch of loguat leaf medicinal materials, measure respectively by above-mentioned sample determination method, the RSD of ursolic acid peak area is 1.56% as a result, shows that the repeatability of this assay method is good.
2.4 testing result: by the need testing solution of 12 batches of loguat leaf medicinal materials of above-mentioned 2.2.1 method preparation, sample detection successively under above-mentioned 2.1 selected same experiment conditions, the result records the HPLC chromatogram of 12 test samples.
2.5 finger-print similarity analysis
2.5.1 the selection at common characteristic peak is analyzed, is compared according to 12 batches of given peak number, peak value (integrated value) and peak position correlation parameters such as (relative retention times) of test sample measurement result, the loguat leaf high performance liquid chromatography is separable to go out more than 10 chromatographic peak, through contrasting with reference substance, and the chromatogram of all mensuration and record relatively, choose the characteristic peak of 8 total peaks as finger-print, wherein No. 8 peaks derive from ursolic acid and (see Fig. 3, A is loguat leaf sample chromatogram figure, and B is a ursolic acid reference substance chromatogram).
2.5.2 the foundation of total figure spectral model imports computer aided similarity evaluation system software with the test data of each batch sample, through selecting the peak, set matching template, will compose the peak and mate automatically, set up its total collection of illustrative plates standard form, carry out the similarity evaluation of each sample then.
2.5.3 the collection of illustrative plates similarity analysis adopts the similarity of each batch of computer aided similarity evaluation system software analysis sample finger-print, the results are shown in Figure 4, table 6～table 8.
Table 6 loguat leaf efficient liquid-phase chromatograph finger print atlas similarity
The relative retention time table of total fingerprint peaks in the table 7 loguat leaf finger-print
The area ratio of total fingerprint peaks in the table 8 loguat leaf finger-print
By analysis, the loguat leaf sample HPLC chromatogram similarity value of 12 batches of different places of production and kind is all greater than 0.9.
Claims (10)
1. the quality evaluating method of a loguat leaf medicinal material comprises the steps:
The first step is set up the standard finger-print of loguat leaf earlier, and key step comprises:
(a) the loguat leaf medicinal material of collection more than 10 batches, and identify its kind and quality;
(b) preparation of loguat leaf medicinal material standard solution;
(c) preparation of object of reference solution;
(d) selection of assay method;
(f) measure: accurate loguat leaf medicinal material standard solution and the object of reference solution drawn, inject detecting instrument respectively, measure the loguat leaf standard finger-print.
In second step, measure loguat leaf sample finger-print to be measured with above-mentioned identical method;
In the 3rd step, with loguat leaf sample finger-print to be measured and the contrast of loguat leaf standard finger-print, screening meets the loguat leaf medicinal material of loguat leaf standard finger-print.
2. loguat leaf quality of medicinal material evaluation method as claimed in claim 1 is characterized in that, the preparation process of described loguat leaf medicinal material standard solution is: get the loguat leaf medicinal material, pulverize, sieve, be dried to constant weight.It is an amount of that precision takes by weighing Loquat Leaf, uses alcohol solvent (the C number is preferably methyl alcohol or normal butyl alcohol less than 5) ultrasonic Extraction 2～3 times, merges extract.Filter, filtrate adds alcohol solvent (the C number is preferably methyl alcohol or normal butyl alcohol less than 5) or moving phase and is diluted to finite concentration after concentrating, and shakes up, promptly.Filter with miillpore filter before the sample introduction.Preferred preparation process is: get the about 1g of Loquat Leaf (crossing sieve No. four), accurate title is fixed, adds 30 times of amount normal butyl alcohols, leaves standstill 15min, ultrasonic Extraction 3 times, each 30min filters, and gets filtrate, and solvent is flung in heating, use dissolve with methanol, move to the 5ml measuring bottle, add methyl alcohol and be diluted to scale, shake up; Promptly.
3. loguat leaf quality of medicinal material evaluation method as claimed in claim 1 is characterized in that object of reference is the effective constituent in the loguat leaf in the preparation of described object of reference solution, is preferably the triterpenic acid compounds, more preferably ursolic acid.
4. loguat leaf quality of medicinal material evaluation method as claimed in claim 1 is characterized in that assay method is a chromatography in the selection of described assay method, is preferably high performance capillary electrophoresis (HPCE) or high performance liquid chromatography (HPLC).
5. as the described loguat leaf quality of medicinal material of claim 1～4 evaluation method, it is characterized in that described loguat leaf standard finger-print is Fujian Province's loguat leaf medicinal materials fingerprint.
6. as the described loguat leaf quality of medicinal material of claim 1～4 evaluation method, it is characterized in that: the finger-print that described employing high-performance capillary electrophoresis method is measured, the relative standard deviation RSD of the relative retention time at its 9 total peaks is all less than 3%, wherein the average relative retention time RT at No. 1 peak is 0.656, and relative standard deviation RSD is 0.62%; The average RT at No. 2 peaks is 0.712, and RSD is 0.50%; The average RT at No. 3 peaks is 0.818, and RSD is 1.08%; The average RT at No. 4 peaks is 0.871, and RSD is 0.34%; The average RT at No. 5 peaks is 0.892, and RSD is 0.30%; The average RT at No. 6 peaks is 0.919, and RSD is 0.29%; The average RT at No. 7 peaks is 0.953, and RSD is 0.62%; No. 8 peaks are ursolic acid, i.e. object of reference, and its average RT is 1.000, RSD is 0.00%; The average RT at No. 7 peaks is 1.070, and RSD is 0.54%.
7. as the described loguat leaf quality of medicinal material of claim 1～4 evaluation method, it is characterized in that: the finger-print of described employing high effective liquid chromatography for measuring, the relative standard deviation RSD of the relative retention time at its 8 total peaks is all less than 3%, wherein the average relative retention time RT at No. 1 peak is 0.322, and relative standard deviation RSD is 2.34%; The average RT at No. 2 peaks is 0.344, and RSD is 2.17%; The average RT at No. 3 peaks is 0.372, and RSD is 1.57%; The average RT at No. 4 peaks is 0.574, and RSD is 2.01%; The average RT at No. 5 peaks is 0.607, and RSD is 1.18%; The average RT at No. 6 peaks is 0.762, and RSD is 2.92%; The average RT at No. 7 peaks is 0.950, and RSD is 1.28%; No. 8 peaks are ursolic acid, i.e. object of reference, and its average RT is 1.000, RSD is 0.00%.
8. as the described loguat leaf quality of medicinal material of claim 1～4 evaluation method, it is characterized in that, the concrete grammar of the preparation of object of reference solution is in the preparation of described object of reference solution: precision takes by weighing ursolic acid reference substance 5mg and places the 10ml volumetric flask, with dissolve with methanol and be diluted to scale, shake up, promptly.
9. as the described loguat leaf quality of medicinal material of claim 1～4 evaluation method, it is characterized in that, the finger-print that described employing high-performance capillary electrophoresis method is measured, its preferred condition is: capillary column (60cm is long, 50 μ m internal diameters); Damping fluid: (0.1mol/L BAS: 0.2mol/L potassium hydroxide solution=4: 2); Separation voltage: 20kV; Experimental temperature: 25 ℃; The detection wavelength is 200nm.Number of theoretical plate calculates by the ursolic acid peak and is not less than 3000.
10. as the described loguat leaf quality of medicinal material of claim 1～4 evaluation method, it is characterized in that, the finger-print of described employing high effective liquid chromatography for measuring, its preferred condition is: chromatographic column: Eclipse XDB-C18 post (4.6mm * 150mm, 5 μ m), moving phase: methyl alcohol: 1% aqueous acetic acid (90: 10), detect wavelength 215nm, flow velocity 1.0mlmin
-1, column temperature is a room temperature, sample size 10 μ l.Number of theoretical plate calculates by the ursolic acid peak and is not less than 2000.
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| CN102370891A (en) * | 2010-08-18 | 2012-03-14 | 浙江中医药大学 | Method for authenticating dendrobium officinale by using HPLC fingerprint |
| CN110873774A (en) * | 2018-08-31 | 2020-03-10 | 山西华康药业股份有限公司 | Method for establishing high performance liquid chromatography fingerprint spectrum of cough-relieving immediate effect pills |
| CN110873774B (en) * | 2018-08-31 | 2022-02-18 | 山西华康药业股份有限公司 | Method for establishing high performance liquid chromatography fingerprint spectrum of cough-relieving immediate effect pills |
| CN110361459A (en) * | 2019-05-20 | 2019-10-22 | 国药集团德众（佛山）药业有限公司 | The method for building up of loguat leaf standard decoction finger-print |
| CN110361459B (en) * | 2019-05-20 | 2022-04-05 | 国药集团德众（佛山）药业有限公司 | Method for establishing loquat leaf standard decoction fingerprint |
| CN114034800A (en) * | 2021-11-24 | 2022-02-11 | 宁夏回族自治区食品检测研究院 | Identification method of medlar producing area and storage period based on multidimensional fingerprint |
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Application publication date: 20090128 |