CN103115973A - Method for judging consistency of traditional Chinese medicine batches - Google Patents

Method for judging consistency of traditional Chinese medicine batches Download PDF

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CN103115973A
CN103115973A CN2013100286076A CN201310028607A CN103115973A CN 103115973 A CN103115973 A CN 103115973A CN 2013100286076 A CN2013100286076 A CN 2013100286076A CN 201310028607 A CN201310028607 A CN 201310028607A CN 103115973 A CN103115973 A CN 103115973A
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chinese medicine
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CN103115973B (en
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聂磊
臧恒昌
曾英姿
王培�
段晓菊
姜红
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Shandong University
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Abstract

The invention discloses a method for judging consistency of traditional Chinese medicine batches. The method comprises the following steps: establishing a reference map on the basis of finger-print similarity analysis, and calculating the mahalanobis distance between a high performance liquid chromatography (HPLC) finger-print in traditional Chinese medicine samples in various batches and the reference map; converting the mahalanobis distance into F check so as to judge whether obvious difference exists between the HPLC finger-print in traditional Chinese medicine samples in different batches and the reference map under the level at a certain probability; and determining whether obvious difference exists in the content of components corresponding to chromatographic peaks in the HPLC finger-print in traditional Chinese medicine samples in various batches by employing a t checking method if the batch samples with obvious difference are determined according to the mahalanobis distance. The method is based on the chromatographic fingerprint and is simple, feasible and practical, the consistency of various batch samples can be judged from the probability according to the difference of the finger-print in traditional Chinese medicine samples in different batches, and method basis is provided for judging the consistency of the batches of the traditional Chinese medicine.

Description

A kind of for Chinese medicine batch conforming determination methods
Technical field
The present invention relates to a kind of for Chinese medicine batch conforming determination methods.
Background technology
Chinese medicine is used for preventing and treating and treating the history of the existing more than one thousand years of disease.Different from synthetic drug, the curative effect performance of Chinese medicine and preparation thereof is mainly the synergy of Multiple components.Nearest decades, due to its hypotoxicity and high curative effect, Chinese medicine is at the demand increase year after year in the whole world, especially in the Asia, Europe and North America.The increase of demand also just means the higher requirement of quality standard proposition.The Chinese medicine of each batch all should satisfy the code requirement of specific products, as date of manufacture and shelf-life etc.Yet even only there is the simple formulations that several herbal medicine are made just may comprise hundreds of uniqueness, narrow spectrum composition, its quality control is many more than the synthetic drug difficulty.If it is that a huge challenge that faces is controlled and estimated to traditional Chinese medicine quality that the contained whole compositions of Chinese medicine are characterized [1]In addition, only screening one or more compositions carries out traditional Chinese medicine quality and controls and do not meet traditional tcm theory [2]In the last few years, by a large amount of studies show that, chromatographic fingerprinting had become a kind of traditional Chinese medicine quality of accepting extensively and had controlled and evaluation method.
1991, WHO promulgated the guilding principle based on traditional Chinese medicine quality, safety, effectiveness and desired use assessment.At that time, chromatographic fingerprinting is used as a kind of acceptable assay method [3]After more than ten years, U.S. food Drug Administration (FDA), EMEA (EMES) and State Food and Drug Administration (SFDA) all accept the quality control that chromatographic fingerprinting is used for herbal drug, and have promoted its development [4～6]As a kind of quality evaluating method, chromatographic fingerprinting can reflect the number of chemical composition information that comprises in complex analyses sample (as Chinese medicine), can provide more comprehensively quality assessment.Control in order to advance fingerprint spectrum method to be effective to traditional Chinese medicine quality, SFDA suggestion chromatographic fingerprints of Chinese materia medica should carry out similarity evaluation and (comprise two kinds of computing method of related coefficient and included angle cosine [7]).Nowadays, similarity has been widely used in the similarity of estimating finger-print.Does yet similarity is higher mean that the difference of finger-print is just less? in recent years, the quality control of Chinese medicine preparation set up and has been used for by high performance liquid chromatography (HPLC) finger-print in conjunction with the content assaying method of some characteristic chemical constituent [8,9]In these research work, satisfactory although the fingerprint similarity of sample is estimated, in different samples, the content of some composition still presents larger difference [10～12]This explanation fingerprint similarity evaluation can not fully reflect the similarity between finger-print, can not prove absolutely the similarity of sample ingredient content.The similarity (or difference) of how estimating between finger-print is a problem that is worth further investigation.Have stability and consistance preferably in order to ensure each batch of Chinese medicine sample quality, the finger-print difference of each batch sample is the smaller the better.Yet, in present stage, for the such complex analyses system of Chinese medicine, realize quality control and unrealistic by its contained all the components being carried out qualitative and quantitative analysis.Therefore, the application has proposed a kind of for traditional Chinese medicine quality batch conforming determination methods based on chromatographic fingerprinting, the method can judge from the angle of probability whether the difference between different batches sample finger-print is remarkable, and can provide that in finger-print, there is larger difference in which chromatographic peak tie element on content, provide a kind of effective, practical method mode to controlling and estimate for traditional Chinese medicine quality.
Summary of the invention
For above-mentioned prior art, the present invention is based on chromatographic fingerprinting, a kind of determination methods for each batch of Chinese medicine unanimity of samples has been proposed, easy, the easy row of the method, practicality can realize the conforming judgement of each batch sample by the difference that judges each batch of Chinese medicine sample finger-print from the angle of probability.The consistance of each batch of Chinese medicine sample can embody by judging the difference between each batch sample fingerprint collection of illustrative plates: if the difference of different batches sample chromatogram finger-print on certain probability level without conspicuousness, the consistance there was no significant difference of each batch sample; Otherwise consistance has significant difference; Batch sample that significant difference is arranged for consistance can analyze that in finger-print, which chromatographic peak differs greatly.
The present invention is achieved by the following technical solutions:
A kind of determination methods for each batch of Chinese medicine unanimity of samples, step is as follows:
(1) select the traditional Chinese medicine sample of a plurality of different batches to be judged, after grinding, adopt the solvent extraction that is fit to extract this Effective Component of Chinese Medicine, obtain extract;
(2) adopt high performance liquid chromatograph at original high efficiency liquid chromatography (HPLC) finger-print of the extract of the traditional Chinese medicine sample of the above-mentioned different batches of detection wavelength place's collection that is fit to this Chinese medicine;
(3) the HPLC finger-print with above-mentioned gained carries out the methodology checking, and (that is: precision test, reappearance test and stability test meet the requirements to make the HPLC finger-print of measuring meet the fingerprint pattern technology code requirement.According to precision test and reappearance testing requirements, investigate the relative retention time of chromatographic peak, the consistance of peak area ratio.Each chromatographic peak of the total peak area ratio of regulation in finger-print, the relative standard deviation of its peak area ratio (RSD) must not be greater than 3%; According to the stability test requirement, investigate the relative retention time of chromatographic peak, the consistance of peak area ratio, determine detection time); Then calculate the average collection of illustrative plates collection of illustrative plates in contrast of HPLC finger-print of the traditional Chinese medicine sample of each batch; Calculate respectively the HPLC finger-print and the similarity that contrasts collection of illustrative plates of the traditional Chinese medicine sample of each batch; If similarity (r) is more than or equal to 0.900, keep, if r＜0.900, it is rejected, then recomputate the contrast collection of illustrative plates of the traditional Chinese medicine sample of residue batch, and the similarity of the contrast collection of illustrative plates of the traditional Chinese medicine sample of the HPLC finger-print of the traditional Chinese medicine sample of each batch of residue and residue batch, and then (r is more than or equal to 0.900 if that is: to adopt criterion similarity same as described above, keep, if r＜0.900, with its rejecting); Repeat this process, until it meets the requirements (that is: similarity r all 〉=0.900);
(4) select the high sample of active constituent content (can select a collection of after above-mentioned selection remaining batch, also can select some batches) the HPLC finger-print as a reference, select the traditional Chinese medicine sample of a plurality of batch nearest with the HPLC finger-print Euclidean distance of reference sample, then build with reference to classification with reference sample, and the average finger-print of computing reference classification sample, namely with reference to collection of illustrative plates;
(5) based on above-mentioned with reference to classification, calculate each batch traditional Chinese medicine sample the HPLC finger-print with reference to the mahalanobis distance of collection of illustrative plates, then mahalanobis distance is converted into F check, thus judge different batches traditional Chinese medicine sample the HPLC finger-print and with reference between collection of illustrative plates, whether there were significant differences under probability level necessarily;
(6) find out the traditional Chinese medicine sample that mahalanobis distance is judged to be those batches of significant difference, adopt the t check to be tested in the total peak of the HPLC finger-print of the traditional Chinese medicine sample of these batches and the total peak with reference to the HPLC finger-print of the traditional Chinese medicine sample of each batch of classification, in judgement total peak, whether each chromatographic peak area is having significant difference under certain probability level; Be judged as the chromatographic peak of significant difference, show in the traditional Chinese medicine sample of this batch, be starkly lower than on the represented active constituent content of this chromatographic peak or higher than the average content of the corresponding composition of other traditional Chinese medicine sample of reference class.
Preferably, described step (4) is specific as follows: select the HPLC finger-print of the high batch sample (reference sample) of active constituent content as a reference, calculate the Euclidean distance of the HPLC finger-print of the HPLC finger-print of traditional Chinese medicine sample of other each batch and reference sample, traditional Chinese medicine sample with other each batch ascending according to Euclidean distance arranged, select the traditional Chinese medicine sample with the HPLC finger-print Euclidean distance of reference sample nearest a plurality of (being no less than 10) batch, and building with reference to classification with reference sample, and computing reference collection of illustrative plates.
Preferably, in described step (5), the consistance quality that judges the traditional Chinese medicine sample of each batch is based on that probable value that mahalanobis distance provides determines; Before calculating mahalanobis distance, elder generation carries out principal component analysis (PCA) (Principal component analysis with reference to the HPLC finger-print data of the traditional Chinese medicine sample of classification, PCA), number of principal components is to determine greater than the principle of All Eigenvalues mean value according to the character pair value.Determine score vector according to number of principal components, and then calculate mahalanobis distance; Then mahalanobis distance is checked by F and provided corresponding probable value (P), if the P of the traditional Chinese medicine sample of certain batch＜0.05(α=0.05), judge this batch traditional Chinese medicine sample the HPLC finger-print with there were significant differences with reference to collection of illustrative plates, represent that there is significant difference in other traditional Chinese medicine sample consistance of this batch sample and reference class.If without significant difference, judge that the traditional Chinese medicine sample of this batch and other traditional Chinese medicine sample of reference class are other batch of same class sample, the consistance there was no significant difference.
In described step (6), adopt the t check to carry out peak area significance test, in theory, chromatographic peak area and the proportional relation of tie element content are consistent based on the t assay of peak area with t assay based on tie element content; In other words, adopt t to upcheck and judge whether peak area has significant difference, can judge that whether tie element content has significant difference, need not the quantitative test of chromatographic peak tie element.
Determination methods for each batch of Chinese medicine unanimity of samples of the present invention, easy, the easy row of method, practicality are suitable for to Chinese medicine batch conforming quick, accurately judgement, for a Chinese medicine batch conforming judgement provides the method foundation.
Description of drawings
The efficient liquid-phase chromatograph finger print atlas of Fig. 1 for adopting U.S. Agilent1200series high performance liquid chromatograph to obtain.
Fig. 2 is for adopting " chromatographic fingerprints of Chinese materia medica similarity evaluation software " to use the similarity evaluation result of each batch sample HPLC finger-print with the contrast collection of illustrative plates of related coefficient or Cosin method calculating.
Fig. 3 is the probability that the different batches sample calculates according to mahalanobis distance with respect to other traditional Chinese medicine sample of reference class.The dotted line that is parallel to horizontal ordinate in figure represents α=0.05 level of significance.
Fig. 4 is the z value that mahalanobis distance is judged to be the 19 batch samples total peak of significant difference.Z wherein CritAccording to t check t under α=0.05 level CritCalculate by formula (2).
Fig. 5 is the frequency number percent that the total peak of significant difference is arranged.
Fig. 6 is the representative HPLC finger-print of Chinese medicine Xin Ke Shu ' tablet for treating coronary heart disease.Wherein the sign chromatographic peak be frequency large percentage shown in Figure 5 (〉 60%) chromatographic peak.
Embodiment
The present invention is further illustrated below in conjunction with embodiment.
Embodiment 1 Chinese medicine used is Xin Ke Shu ' tablet for treating coronary heart disease, and sample batch is 71 batches.Xin Ke Shu ' tablet for treating coronary heart disease mainly is comprised of the red sage root, the root of kudzu vine, pseudo-ginseng, hawthorn and the banksia rose as the Chinese patent drug of clinical treatment coronary disease and angina pectoris commonly used.The method of the invention is to Chinese medicine used and unrestricted.
1.1 the preparation of need testing solution
At first get the Xin Ke Shu ' tablet for treating coronary heart disease sample of appropriate different batches (this example is 71 batches), remove the outer coatings film, grind.Precision takes ground sample fine powder 100mg, adds 70% ethanol (percent by volume) 5mL, and ultrasonic extraction 20min under room temperature gets 4mL at the centrifugal 10min of 3000r/min.Then get supernatant after 0.45 μ m membrane filtration, sample introduction adopts the efficient liquid phase chromatographic analysis instrument to record can the relax original HPLC finger-print of ethanol extract composition of these different batches hearts, as shown in Figure 1.
1.2 instrument and chromatographic condition
Agilent1200 liquid chromatograph (Agilent Corp., Santa Clara, CA, USA), DAD detecting device, PhenomexC 18(250mm * 4.6mm, 4 μ m); Mobile phase: acetonitrile (A) and 0.1%(percent by volume) formic acid (B) aqueous solution; Gradient elution: 0-21min, 8-12% (A); 21-31min, 12-17% (A); 31-55min, 17-38% (A); 55-65min, 38-90% (A); 65-75min, 90% (A), flow velocity are 1.0ml/min, and sample size is 10 μ L, and column temperature is 30 ℃.The detection wavelength is 278nm.
1.3 the methodology of finger-print checking
With same batch of sample extracting solution continuous sample introduction 6 times, the relative retention time at the finger-print of mensuration total peak and the relative standard deviation (RSD) of peak area ratio are respectively less than 1.2% and 3.0%, show that the precision of method is better.Core 5 parts, Xin Ke Shu' tablet sample, obtain 5 duplicate samples extracts by 1.1 described methods, measure the HPLC finger-print in accordance with the law, the relative retention time at total peak and the RSD of peak area ratio are respectively less than 1.4% and 1.8%, show the repeatability of method better, meet the technical requirement of finger-print.Same batch of sample extracting solution carried out 6 times measure in 3 days, get the RSD of the relative retention time at HPLC finger-print total peak and peak area ratio respectively less than 2.3% and 3.0%, show that sample extracting solution is stable in 3 days.
1.4 similarity analysis
Adopt " chromatographic fingerprints of Chinese materia medica similarity evaluation software " (2004A version, pharmacopoeia commission's recommendation) the HPLC finger-print data of obtaining are carried out Supplements, coupling generates the contrast collection of illustrative plates, obtains the data matrix of each batch sample HPLC finger-print.In this matrix, " OK " expression sample, " row " expression variable (namely chromatographic peak).Calculate respectively each batch sample HPLC finger-print and the similarity that contrasts collection of illustrative plates, result such as Fig. 2 by related coefficient and Cosin method.As seen the similarity value of each batch sample HPLC finger-print of two kinds of method calculating is very high, all greater than 0.900.
1.5 other structure of reference class
Select the high batch sample of active constituent content as the reference sample.It is reference sample that batch sample 5 is adopted in this experiment, because of its puerarin content maximum in each batch sample.Based on reference sample HPLC finger-print, employing Matlab software calculates the Euclidean distance of each batch sample HPLC finger-print and reference sample HPLC finger-print, with the big or small ascending arrangement of each batch sample by Euclidean distance.Select the less batch sample of some and reference sample HPLC finger-print Euclidean distance to build with reference to classification.Other sample size of reference class can be determined by the user.The technical requirement of this experimental basis finger-print research, namely finger-print sample batch quantity is no less than 10 batches and is foundation, determine reference class very product batch quantity be 11, comprise reference sample (being sample 5).
1.6 each batch sample consistance judgement
Adopt the average finger-print (namely with reference to collection of illustrative plates) of Matlab software computing reference classification sample HPLC finger-print, then calculate all batches sample HPLC finger-print and mahalanobis distance (Mahalanobis distance) with reference to collection of illustrative plates.Before calculating mahalanobis distance, need first to carry out principal component analysis (PCA) (Principalcomponent analysis, PCA) with reference to classification sample HPLC finger-print data.Because the variable number (being the chromatogram peak number) of reference class other style product HPLC finger-print data is greater than sample number, such matrix can't directly carry out mahalanobis distance and calculate.In the PCA process, number of principal components is to determine greater than the principle of the mean value of All Eigenvalues according to the character pair value.In this example, the eigenwert of front 2 major components is greater than the average of All Eigenvalues, so number of principal components is defined as 2.Reference class very product HPLC finger-print data obtains load vector (being proper vector) through PCA, then calculate the score of all batches sample HPLC finger-print data according to load vector, so calculate again each batch sample HPLC finger-print with reference to the mahalanobis distance of collection of illustrative plates.The mahalanobis distance that calculates is checked by F calculated corresponding probable value (P), judge each batch sample HPLC finger-print and with reference to collection of illustrative plates (α=0.05) under certain level of significance, whether significant difference arranged with this.If P＜0.05(α=0.05), corresponding batch sample HPLC finger-print with there were significant differences with reference to collection of illustrative plates, illustrates this batch sample and has significant difference with reference to a classification batch sample consistance.If P 〉=0.05(α=0.05), corresponding batch sample HPLC finger-print with reference to collection of illustrative plates without significant difference, show this batch sample with reference to classification batch sample consistance there was no significant difference.The different batches sample is seen Fig. 3 with respect to reference class other style product by the probability that mahalanobis distance calculates, and wherein front 11 samples are very product of reference class.By shown in Figure 3, under α=0.05 level, reference class is product HPLC finger-print and there is no significant difference with reference to collection of illustrative plates very.In other batch sample, there are 19 batches of samples (being Fig. 3 the right) with respect to the reference collection of illustrative plates, significant difference (namely reference class other style product are overall) to be arranged, these batches sample finger-print is described and may has larger difference with reference to collection of illustrative plates.
Mahalanobis distance is used for judging each batch sample HPLC finger-print and with reference to whether significant difference is arranged between collection of illustrative plates, and in there were significant differences batch sample, which finger-print chromatographic peak area has significant difference but be difficult to find out.In this example, adopt the t method of inspection can judge which chromatographic peak area has significant difference.Be the common characteristic of each batch sample because finger-print has the peak, the area discrepancy of these chromatographic peaks is particularly important for batch sample consistance.For this reason, adopt the t method of inspection that the total peak area of batch sample HPLC finger-print that mahalanobis distance is judged to be significant difference is tested, judge which total peak area and with reference to the total peak area of classification batch sample HPLC finger-print, significant difference (α=0.05) arranged.In the t checkout procedure, chromatographic peak area need to be transformed by formula (1),
z i = x i - x ‾ i S i - - - ( 1 )
In formula (1), x iRefer to the peak area of certain batch sample HPLC finger-print chromatographic peak i.
Figure BDA00002775233500062
And S iBe respectively peak area mean value and standard deviation with reference to each batch sample HPLC finger-print chromatographic peak i in classification.With z iValue is calculated t by formula (2) i:
t i＝z i[n/(n+1)] 1/2 （2）
In formula (2), n represents very product quantity of reference class.t iKey value (t with student t distribution Crit, i.e. α=0.05 o'clock corresponding t value) compare, to determine x iWith
Figure BDA00002775233500063
Whether significant difference is arranged.By formula (1) and (2) as can be known, although t iBe based on that the area of finger-print chromatographic peak i calculates, but it is consistent with result of calculation based on chromatographic peak i tie element content.Proof procedure is as follows:
In chromatographic quantitative analysis, the area of a certain chromatographic peak i and the following relation of the content of tie element, as the formula (3),
x i＝f ic i+b i （3）
C wherein iThe concentration of expression chromatographic peak i tie element, f iAnd b iBe regularly constant at chromatographic condition one.Formula (3) substitution formula (1) is had
z i = ( x i - x ‾ i ) / s i = f i ( c i - c ‾ i ) f i [ 1 n - 1 Σ i = 1 n ( c i - c ‾ i ) 2 ] 1 / 2 = ( c i - c ‾ i ) / s c i - - - ( 4 )
By formula (4) as seen, based on the z of calculated by peak area iWith the z based on the cubage of chromatographic peak tie element iIdentical, so t i(seeing formula (2)) is also identical, and card is finished.
By above-mentioned proof as can be known, be consistent based on the t assay of chromatographic peak area with t assay based on chromatographic peak tie element content.Therefore, we can just can judge that by the t assay based on chromatographic peak area whether chromatographic peak i tie element content in certain batch of sample HPLC finger-print and average content with reference to classification batch sample composition i have significant difference, need not in advance component i to be carried out quantitative test.In certain batch of sample HPLC finger-print, which chromatographic peak tie element content whether have significant difference very convenient to this result for investigating, make and adopt other method (as mass spectrophotometry) that composition is done to identify that further Objective is stronger, for determining that affecting a Chinese medicine batch conforming factor has important directive significance.
In this example, when adopting the t check to carry out significance test to total peak, with the t of α=0.05 o'clock CritThrough type (2) is converted into z Crit, then directly compare to judge with the z value that adopts formula (1) to calculate based on peak area.The total peak z value that is judged as 19 batch samples of significant difference by mahalanobis distance is seen Fig. 4.As shown in Figure 4, as long as z iZ Crit, illustrate that chromatographic peak i is judged to be the total peak of significant difference through the t check.The total peak area that in Fig. 6, major part has a significant difference is less all, illustrates that the content of tie element may be lower than the reference class average content of the corresponding composition of product very.Frequency number percent that the total peak of significant difference occurs in 19 batch samples also inconsistent (as shown in Figure 5) is arranged.Wherein, the ratio of No. 13 and No. 19 total peak appearance is the highest, is secondly No. 16 and No. 20 total peaks.The total peak (〉 60% of appearance large percentage) retention time in the HPLC finger-print as shown in Figure 6.
As above embodiment shows, the present invention can judge a Chinese medicine batch consistance by mahalanobis distance on the basis that fingerprint similarity is analyzed, whether significantly can provide batch difference of sample room from the angle of probability.Be judged to be for mahalanobis distance batch sample that there were significant differences, adopt the t method of inspection can judge in these batches sample HPLC finger-print, which chromatographic peak tie element content whether have significant difference.
Embodiment recited above limits design of the present invention and protection domain, and various modification and improvement that those skilled in the art make technical scheme of the present invention all should fall into protection scope of the present invention.
List of references
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Figure BDA00002775233500081
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Claims (3)

1. determination methods that is used for each batch of Chinese medicine unanimity of samples, it is characterized in that: step is as follows:
(1) select the traditional Chinese medicine sample of a plurality of different batches to be judged, after grinding, adopt the solvent extraction that is fit to extract this Effective Component of Chinese Medicine, obtain extract;
(2) adopt high performance liquid chromatograph at the original high efficiency liquid-phase chromatograph finger print atlas of the extract of the traditional Chinese medicine sample of the above-mentioned different batches of detection wavelength place's collection that is fit to this Chinese medicine;
(3) the HPLC finger-print with above-mentioned gained carries out the methodology checking, makes the HPLC finger-print of measuring meet the fingerprint pattern technology code requirement; Then calculate the average collection of illustrative plates collection of illustrative plates in contrast of HPLC finger-print of the traditional Chinese medicine sample of each batch; Calculate respectively the HPLC finger-print and the similarity that contrasts collection of illustrative plates of the traditional Chinese medicine sample of each batch; If similarity r is more than or equal to 0.900, keep, if r＜0.900, with its rejecting, then recomputate the contrast collection of illustrative plates of traditional Chinese medicine sample of residue batch, and the similarity of contrast collection of illustrative plates of the traditional Chinese medicine sample of the HPLC finger-print of the traditional Chinese medicine sample of each batch of residue and residue batch, and then adopt criterion similarity same as described above; Repeat this process, until it meets the requirements;
The HPLC finger-print of the sample that (4) remaining batch, the selection active constituent content is high after above-mentioned selection as a reference, select the traditional Chinese medicine sample of a plurality of batch nearest with the HPLC finger-print Euclidean distance of reference sample, then build with reference to classification with reference sample, and the average finger-print of computing reference classification sample, namely with reference to collection of illustrative plates;
(5) based on above-mentioned with reference to classification, calculate each batch traditional Chinese medicine sample the HPLC finger-print with reference to the mahalanobis distance of collection of illustrative plates, then mahalanobis distance is converted into F check, thus judge different batches traditional Chinese medicine sample the HPLC finger-print and with reference between collection of illustrative plates, whether there were significant differences under probability level necessarily;
(6) find out the traditional Chinese medicine sample that mahalanobis distance is judged to be those batches of significant difference, adopt the t check to be tested in the total peak of the HPLC finger-print of the traditional Chinese medicine sample of these batches and the total peak with reference to the HPLC finger-print of the traditional Chinese medicine sample of each batch of classification, in judgement total peak, whether each chromatographic peak area is having significant difference under certain probability level; Be judged as the chromatographic peak of significant difference, show in the traditional Chinese medicine sample of this batch, be starkly lower than on the represented active constituent content of this chromatographic peak or higher than the average content of the corresponding composition of other traditional Chinese medicine sample of reference class.
2. a kind of determination methods for each batch of Chinese medicine unanimity of samples according to claim 1, it is characterized in that: described step (4) is specific as follows: select the HPLC finger-print of the high batch sample of active constituent content as a reference, calculate the Euclidean distance of the HPLC finger-print of the HPLC finger-print of traditional Chinese medicine sample of other each batch and reference sample, traditional Chinese medicine sample with other each batch ascending according to Euclidean distance arranged, select the traditional Chinese medicine sample of a plurality of batch nearest with the HPLC finger-print Euclidean distance of reference sample, and build with reference to classification with reference sample, and computing reference collection of illustrative plates.
3. a kind of determination methods for each batch of Chinese medicine unanimity of samples according to claim 1 is characterized in that: in described step (5), the consistance quality that judges the traditional Chinese medicine sample of each batch is based on that probable value that mahalanobis distance provides determines; Before calculating mahalanobis distance, elder generation carries out principal component analysis (PCA) with reference to the HPLC finger-print data of the traditional Chinese medicine sample of classification; Determine score vector according to number of principal components, and then calculate mahalanobis distance; Then mahalanobis distance is checked by F and provided corresponding probable value P, if the P of the traditional Chinese medicine sample of certain batch＜0.05, α=0.05, judge this batch traditional Chinese medicine sample the HPLC finger-print with there were significant differences with reference to collection of illustrative plates, represent that there is significant difference in other traditional Chinese medicine sample consistance of this batch sample and reference class; If without significant difference, judge that the traditional Chinese medicine sample of this batch and other traditional Chinese medicine sample of reference class are other batch of same class sample, the consistance there was no significant difference.
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CN106109845A (en) * 2016-05-05 2016-11-16 中国医学科学院药用植物研究所云南分所 For treating the Sanguis Draxonis spray of pressure ulcer and inspection thereof and preparation method
CN106109845B (en) * 2016-05-05 2019-07-26 中国医学科学院药用植物研究所云南分所 For treating the Resina Draconis spray and its inspection and preparation method of pressure sore
CN106690401A (en) * 2017-03-08 2017-05-24 云南中烟工业有限责任公司 Cigarette formula quality trend analysis method based on volatile characteristic components in cut tobacco
CN106690401B (en) * 2017-03-08 2018-01-30 云南中烟工业有限责任公司 Cigarette composition quality trends analysis method based on volatility characteristics component in pipe tobacco
CN111562337A (en) * 2019-02-13 2020-08-21 中国石油天然气股份有限公司 Method and system for identifying polymer product
CN110095429A (en) * 2019-04-30 2019-08-06 济南弗莱德科学仪器有限公司 A kind of product oil method for quickly detecting quality
CN113029979A (en) * 2021-02-10 2021-06-25 河南中烟工业有限责任公司 Method for testing quality stability of cigarette paper
CN116973495A (en) * 2023-09-21 2023-10-31 山东鲁地源天然药物有限公司 Analysis and management system for detection data of traditional Chinese medicine decoction pieces based on gas chromatograph
CN116973495B (en) * 2023-09-21 2023-12-15 山东鲁地源天然药物有限公司 Analysis and management system for detection data of traditional Chinese medicine decoction pieces based on gas chromatograph

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