CN116124911A

CN116124911A - Quality evaluation method for Chinese patent medicine based on multisource information fusion

Info

Publication number: CN116124911A
Application number: CN202210931998.1A
Authority: CN
Inventors: 林永强; 许丽丽; 崔伟亮; 焦阳; 郭东晓; 穆向荣; 薛菲; 尹雪; 徐兴燕; 张建刚
Original assignee: Shandong Institute for Food and Drug Control
Current assignee: Shandong Institute for Food and Drug Control
Priority date: 2022-08-04
Filing date: 2022-08-04
Publication date: 2023-05-16

Abstract

The invention discloses a quality evaluation method of a Chinese patent medicine based on multisource information fusion, which comprises the following steps: 1) Testing according to standard and non-standard methods; 2) Taking quantifiable items with differences as evaluation indexes; 3) Determining the weight W and the assigned value A of the item; 4) Calculating the score of the evaluation item according to the assigned value and the weight coefficient; 5) According to

And sorting according to the scores from high to low so as to evaluate the relative quality. The invention uses NLSC as a research object, integrates multi-source information such as traditional microscopic identification, fingerprint spectrum, LC-MS/MS, proteomics, molecular biology identification technology and the like, adopts a weight evaluation method of information fusion, and constructs a new quality control of Chinese patent medicineAnd (3) a strategy for proposing and implementing the quality evaluation standard of the traditional Chinese medicine which is accepted in the pharmaceutical industry and even the world.

Description

Quality evaluation method for Chinese patent medicine based on multisource information fusion

Technical Field

The invention belongs to the field of traditional Chinese medicine quality control and evaluation methods, and particularly relates to a quality evaluation method of a Chinese patent medicine based on multi-source information fusion.

Background

The Chinese medicine is a characteristic of Chinese medicine and is an important component of Chinese traditional culture. Plays an irreplaceable role in world medical and health activities. The modernization of traditional Chinese medicine is to develop a modern traditional Chinese medicine product with high efficiency, safety, stability and controllable quality by applying modern advanced scientific technology and following international standards and specifications on the basis of inheriting and developing the advantages and characteristics of traditional Chinese medicine. In recent years, modernization of traditional Chinese medicine still has the following challenges: 1) The advantages of the traditional microscopic identification are not fully reflected; 2) Only one or two index components cannot represent the overall chemical characteristics of the Chinese patent medicine; 3) All aspects of the information are isolated, cannot be effectively integrated, and cannot form a combination force. Therefore, development of a multi-source information fusion strategy based on combination of traditional experience identification and modern instrument analysis is urgently needed, quantitative evaluation is carried out on medicine quality according to multiple indexes and factors, the quality sequence is discharged, quality of different enterprises and different batches of Chinese medicine varieties is effectively evaluated, and a new path for evaluating Chinese medicine quality is constructed.

The Naolingsu capsule is a widely used Chinese medicine compound prescription containing 15 kinds of Chinese herbal medicines and is used for treating neurasthenia, amnesia, insomnia, dizziness, palpitation, physical fatigue, impotence and spermatorrhea. In the prescription, rhizoma polygonati, epimedium and shizandra berry are used as monarch drugs, and have the effects of tonifying kidney and strengthening yang, dispelling wind and removing dampness, and strengthening body and bones. Fructus Xanthii and fructus Lycii have remarkable pharmacological properties, and are beneficial for age-related diseases, immune diseases, etc. Ginseng radix and semen Ziziphi Spinosae have antioxidant, blood vessel and heart function improving, immunity enhancing and anxiolytic effects, and can be used for treating insomnia and palpitation. Radix Ophiopogonis, carapax et Plastrum Testudinis, and rehmanniae radix have effects of invigorating kidney, nourishing yin, and replenishing blood. The seven medicinal herbs are used as ministerial drugs. Poria, fructus Jujubae, cornu Cervi Pantotrichum and colla Cornus Cervi are adjuvants in the prescription. Polygala tenuifolia is commonly used as a guiding drug for the treatment of insomnia, memory impairment and neurasthenia. The whole formula has the effects of tonifying qi, nourishing blood, strengthening brain and soothing nerves.

However, the quality standard of Naolinsu capsules is carried in the 8 th volume (standard number WS 3-B-3092-98) of traditional Chinese medicine prescription preparation, and only characters and conventional inspection items are used as the guarantee of effectiveness and safety, and the pharmacodynamics substance basic research and quality control of Naolinsu capsules are quite limited so far.

Disclosure of Invention

The invention aims to provide a quality evaluation method of a Chinese patent medicine based on multi-source information fusion, which realizes qualitative and quantitative detection of multiple medicines in a prescription and even full medicines, introduces innovation by standard, promotes high-quality sustainable development of the traditional Chinese medicine industry, and provides reference basis for national and provincial traditional Chinese medicine evaluation sampling inspection work.

In order to achieve the above purpose, the present invention is realized by the following technical scheme:

a quality evaluation method of Chinese patent medicine based on multisource information fusion comprises the following steps:

(1) Detection is carried out according to standard and non-standard methods, wherein the non-standard methods comprise microscopic identification, HPLC-DAD chromatography, LC-MS/MS chromatography, chemometric analysis, deer antler specificity detection (LC-ESI-MS analysis), similarity evaluation, multi-index content measurement, deer antler feeding project measurement, and the standard method is standard number WS ₃ -B-3092-98；

(2) Finding out quantifiable items with differences in the detection results of the two methods as evaluation indexes according to the detection results of the step (1); specifically, components with VIP (contribution value) more than 1 and a pilose antler feeding item in microscopic identification, similarity evaluation and multi-index content measurement are set as evaluation indexes i;

(3) Determining that the weight W of the evaluation index in the step (2) is 0.9 or 1.0, wherein the weight coefficient of the evaluation index with the failure rate lower than 50% is determined to be 1.0, and the failure rate higher than 50% is determined to be 0.9; determining the limit value of the evaluation index in the step (2) to be 80-100 respectively, and calculating the assigned value A by combining the specific conditions of each index _i ；

(4) Calculating the score X of the evaluation item according to the assigned value and the weight coefficient in the step (3) _i ＝A _i W _i The method comprises the steps of carrying out a first treatment on the surface of the i is 1 to 10;

(5) Score X of the evaluation item _i The SPSSAU selection entropy TOPSIS method is introduced to calculate the contribution coefficient S of each evaluation item _i ；

(6) According to

And sorting according to the scores from high to low so as to evaluate the relative quality.

The invention has the beneficial effects that:

the invention takes NLSC as a research object, integrates multi-source information such as traditional microscopic identification, fingerprint spectrum, LC-MS/MS, proteomics, molecular biology identification technology and the like, adopts a weight evaluation method of information fusion, avoids the defect of subjective weighting, constructs a new quality control strategy of the Chinese patent medicine, and proposes and implements the quality evaluation standard of the Chinese patent medicine accepted in the pharmaceutical industry even the world.

Drawings

Other features, objects and advantages of the present invention will become more apparent upon reading of the detailed description of non-limiting embodiments, given with reference to the accompanying drawings in which:

FIG. 1 is a schematic illustration of a microidentification of Naolingsu capsules;

FIG. 2 is a fingerprint of Naolinsu capsule;

FIG. 3 is a graph of PLS-DA scores for 24 samples of Naolin capsules;

FIG. 4 is a graph of predicted variable importance for 24 batches of samples;

FIG. 5 shows the secondary mass spectrum of cornu Cervi Pantotrichum polypeptide A and the assignment of y ion and b ion.

Detailed Description

The invention is further described in connection with the following detailed description, in order to make the technical means, the creation characteristics, the achievement of the purpose and the effect of the invention easy to understand.

(1) The invention discloses a quality evaluation method of a Chinese patent medicine based on multisource information fusion, which comprises the following steps: according to standard (Standard number WS) ₃ -B-3092-98) and non-standard methods (microscopic identification, HPLC-DAD chromatography, LC-MS/MS chromatography, chemometric analysis, deer antler specificity examination, deer antler feed-program detection, etc.);

(2) Finding out quantifiable items with differences in the detection results of the two methods as evaluation indexes according to the detection results of the step (1); specifically, microscopic identification (i=1), similarity evaluation (i=2), composition RD, MPA, DS, SSB, EC, ICA, JB (i=3-9) of VIP (contribution value) > 1 in multi-index content measurement, and a deer antler feed item (i=10) were set as evaluation indexes;

(3) Determining that the weight W of the evaluation index in the step (2) is 0.9 or 1.0 (the weight coefficient of the evaluation index with the failure rate lower than 50% of a certain item is determined to be 1.0, and the failure rate higher than 50% is determined to be 0.9); determining the limit value of the evaluation index in the step (2) to be 80-100 respectively, and calculating the assigned value A by combining the specific conditions of each index _i ；

Specifically, the microscopic identification project is set to be 80 (each medicinal material occupies 10 points) in total, and the weight coefficient is 0.9. The assigned value is combined with the condition A of drug flavor feeding ₁ 60 to 80. The total score for similarity evaluation was set to 100 points and the weight coefficient was 0.9. Similarity item score A ₂ =similarity result×100. Seven components with contribution value more than 1 are obtained through chemometric analysis in multi-index content measurement, and RD (from rehmannia root), MPA (from dwarf lilyturf tuber), DS (from polygala tenuifolia), SSB (from shizandra berry) and JB (from wild jujube seed) can reflect the feeding condition of a certain medicine taste and are all set to 100 minutes. ICA and EC are derived from Epimedium herb and are divided into 100 points. The ICA and JB weight coefficients are 1.0,RD, MPA, DS, SSB and EC are weighted by 0.9. Content measurement item A _i(i＝3-9) =assay result/average content,

the feeding set score of the pilose antler is 100, and the weight coefficient is 1. If no feed was added, the score was 0. Namely the value A of

pilose antler

₁₀ 0 or 100, respectively.

(4) Calculating score X of evaluation item _i ＝A _i W _i ；

(5) Score X of the evaluation item _i Importing an SPSSAU selection entropy TOPSIS method, and calculating a contribution coefficient S of each evaluation item;

(6) According to

The method is verified in the quality standard of Naolingsu capsules:

1. instrument and reagent

Waters 2695 high Performance liquid chromatography System (Waters Inc. of USA) AB sciex 6500 ⁺ High performance liquid chromatography mass spectrometry (shimadzu corporation); XSE205 parts per million electronic balance (METTLER company); BK-600C ultrasonic cleaner (Baker ultrasonic equipment Co., ltd.).

The reference substances chlorogenic acid, 3,6' -sinapioyl sucrose, epimedin C, icariin, schizandrin A, baohuoside I, schizandrin A, schizandrin B, spinosyn D, ginsenoside Rf, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and polygala root oral mountain ketone III are purchased from Chinese food and drug verification institute.

Methyl ophiopogon flavanone A, dehydro-deluxe, polyporus acid C, thiazine diketone glycoside, thiazine diketone are purchased from Chengdu Poisson biotechnology Co., ltd; epimedin A, epimedin B, neochlorogenic acid, schizandrin C and pachymic acid controls were purchased from Chengdoman Biotechnology Co., ltd.

The samples were 24 batches (A1-A2, B1-B4, C1-C7, D1, E1-E10) from 5 enterprises in 2021 national evaluation laboratory.

Acetonitrile (Honeywell B & J) and phosphoric acid (shanghai aladine biotechnology limited)) were both chromatographically pure; the rest reagents are all analytically pure (Tianjin Cord. Miou chemical reagent Co., ltd.); the water was Milli-Q ultra pure water (Millipore co.).

On-line data statistics analysis software SPSSAUSPSSAU-on-line SPSS analysis software)。

2. Microscopic identification

Powder properties of NLSC (Naolinsu Capsule) were identified as the first step of identification and characterization. NLSC powder was taken and observed under a microscope (FIG. 1, magnification X400): the irregular branched block is colorless and is dissolved in chloral hydrate liquid; the mycelium is colorless or pale brown and has a diameter of 4-6 μm (Poria cocos, 1a,1 b). Stone cells are square or rectangular, have a diameter of 30-64 mu m, are thicker in wall, and are thin on one side in some cases, and are fine in tattooing holes; the irregular block is grey yellow, and has micro-texture or pores (tortoiseshell, 2a, 2 b) on the surface. The wood plug cells are light yellow or nearly colorless, polygonal, square-like or long-like, the vertical peripheral wall is thicker, and the holes are intermittent (polygala root, 3). The seed coat grating cells are brownish red, the surface of the seed coat grating cells is polygonal, the diameter of the seed coat grating cells is about 15 mu m, the wall thickness of the seed coat grating cells is lignified, and the cells are small; the side surface is in a strip shape, the outer wall is thickened, the upper part and the middle part of the side wall are very thick, and the lower part is gradually thinned; the bottom surface is similar to a polygon or a round polygon (semen Ziziphi Spinosae, 4a, 4b, 4 c). The surface of the stone seed cells is irregularly polygonal, has a thick wall, is wavy and is bent, and the layering is clear (medlar, 5a and 5 b). The parenchyma contains a plurality of large mucus cells and calcium oxalate needle crystal bundles (rhizoma polygonati, 6a,6 b). The surface of the seed coat epidermic stone cell is polygonal or long polygonal, the diameter is 18-50 mu m, the wall thickness is thick, the pore canal is extremely fine, and the cell cavity contains dark brown matters; the seed coat inner layer stone cells are polygonal, quasi-circular or irregular, have a diameter of about 83 μm, have a slightly thicker wall and larger pits (schisandra chinensis, 7a,7 b). The endothelial cell wall is uniformly thickened; the stone cells have thickened inner and outer walls and even and fine pits (radix Ophiopogonis, 8a, 8 b).

According to the current standard Chinese medicine formulation volume 8 (standard number WS 3-B-3092-98), powder identification of NLSC cannot be realized. The eight kinds of medicine in the prescription can be qualitatively identified through microscopic identification,in particular, tortoise shell, sealwort and medlar have no obvious chemical component identification characteristics. Therefore, the research supplements and perfects the qualitative identification of NLSC, and is more visual and clear. The project reject batch was 33.3% and less than 50% by analysis of all samples, thus setting the weight coefficient to 0.9. In summary, the microscopic identification project is set to be evaluated to be 80 (each medicinal material occupies 10 minutes), and the weight coefficient is 0.9. The microscopic characteristics of the enterprise preparations (4 batches) can be detected, and the assigned value is 80; C. the microscopic features of dwarf lilyturf tuber and tortoise shell in the preparation (8 batches) of the enterprise are difficult to observe, the feeding condition can be indirectly reflected, and problems exist, and the scoring value is 60 points. According to X _i ＝A _i W _i Calculating the score of the evaluation item;

3. finger print

3.1HPLC-DAD chromatography conditions

Chromatographic column: hypersil Gold C ₁₈ (250 mm. Times.4.6 mm,5 μm); acetonitrile (A) -0.1% phosphoric acid aqueous solution (B) is taken as a mobile phase, and gradient elution is carried out (0-5 min,5% A, 5-20 min, 5%. Fwdarw.9% A, 20-50 min, 9%. Fwdarw.40% A, 50-60 min, 40%. Fwdarw.50% A, 60-70 min, 50%. Fwdarw.75% A, 70-80 min, 75%. Fwdarw.95% A, 80-85 min,95% A); volume flow 1.0 mL/min ^-1 Switching detection wavelength detection (0-40 min,326nm, 40-50 min,268nm, 50-85 min,210 nm); sample injection amount: 10 mu L.

3.2HPLC-DAD Mixed control solution Each appropriate amount of the novel chlorogenic acid, 3,6' -sinapiyl sucrose, epimedin A, epimedin B, epimedin C, icariin, schizandrin A, baohuoside I, schizandrin A and schizandrin B of the control is precisely weighed, and is dissolved by adding methanol to prepare mixed control solutions of 3.93 mug/mL, 2.49 mug/mL, 20.52 mug/mL, 10.18 mug/mL, 8.80 mug/mL, 7.59 mug/mL, 23.21 mug/mL, 43.47 mug/mL, 6.68 mug/mL, 7.03 mug/mL and 8.18 mug/mL.

3.3 taking a proper amount of the content of the Naolingsu capsule from the sample solution, grinding, taking about 1.0g, precisely weighing, placing into a conical flask with a plug, precisely adding 25mL of 70% methanol solution, weighing, performing ultrasonic treatment (power is 250W and frequency is 40 kHz) for 30min, taking out, cooling, supplementing the loss with 70% methanol solution, shaking uniformly, and filtering with a 0.22 mu m filter membrane to obtain the Naolingsu capsule.

3.4 establishment of fingerprint and similarity evaluation

HPLC-DAD fingerprint was established by optimizing the chromatographic conditions (refer to chromatographic conditions under 2.1) of NLSC active ingredient. The common peak was determined by localization of the standard and control substances and mass spectrometry. 25 common peaks in the common pattern were co-labeled (FIG. 2).

Peak

3, 5, 8, 9, 10, 11, 12, 13, 14, 21, 23 are neochlorogenic acid, chlorogenic acid, 3,6' -sinapiyl sucrose, epimedin A, epimedin B, epimedin C, icariin, schizandrin A, baohuoside I, schizandrin A, schizandrin B, respectively. And quantitatively determining the contents of the 11 components.

The HPLC-DAD method analyzed fingerprint similarity ranges for 24 batches of NLSCs of 0.969, 0.961, 0.966, 0.959, 0.963, 0.967, 0.976, 0.974, 0.972, 0.971, 0.979, 0.866, 0.877, 0.645, 0.977, 0.987, 0.989, 0.977, 0.992, 0.987, 0.985, 0.991, 0.985, 0.972. The similarity evaluation is that only one batch of samples is less than 0.85, and the reject sample of the project accounts for 4.2%, so that the weight coefficient is determined to be 0.9.

The total score of the similarity evaluation was set to 100 points, and the weight coefficient was 0.9.

Similarity X _i =similarity result×100×0.9.

4. HPLC-DAD and LC-MS/MS methods simultaneously determine 25 index components

4.1HPLC-DAD chromatography conditions under the same "3.1" term

4.2HPLC-DAD Mixed control solution under the heading "3.2

4.3LC-MS/MS chromatographic conditions: chromatographic column: waters ACQUITY UPLC HSS C18 (2.1X100 mm,1.8 μm); methanol-acetonitrile (1:1) is taken as a mobile phase A, 0.1% formic acid solution is taken as a mobile phase B, and gradient elution is carried out: 0-6 min,16% A; 6-11 min,35% A; 11-14 min, 35-65% A. The flow rate was 0.3ml/min and the column temperature was 40 ℃.

Mass spectrometry conditions: electrospray negative ion mode (ESI) using a mass spectrometer detector ^- ) Multiple Reaction Monitoring (MRM) was performed and specific information for each component is shown in Table 1.

TABLE 1 LC-MS/MS specific detection parameters for each component ^q product ion used for quantification

4.4LC-MS/MS method mixing control solution

The 14 kinds of reference substances are weighed respectively, and 70% methanol is added to prepare a mixed reference substance stock solution containing 2 mug of the reference substances per milliliter.

4.5 taking a proper amount of the content of the Naolingsu capsule from the sample solution, grinding, taking about 1.0g, precisely weighing, placing into a conical flask with a plug, precisely adding 25mL of 70% methanol solution, weighing, performing ultrasonic treatment (power is 250W and frequency is 40 kHz) for 30min, taking out, cooling, supplementing the loss with 70% methanol solution, shaking uniformly, and filtering with a 0.22 mu m filter membrane to obtain the Naolingsu capsule.

4.6 methodological verification

The HPLC-DAD and LC-MS/MS methods simultaneously measure 25 index components according to the principle of Chinese pharmacopoeia methodology verification and guidance.

Table 2 methodological validation results

/>

^a μg/ml(data acquired by HPLC-DAD)

^b ng/ml(data acquired by LC-MS/MS)

4.7 chemometric analysis

4.7.1 principal component analysis

And (3) normalizing the content measurement results of 25 index components in 24 batches of NLSCs by adopting SIMCA-P software, and calculating the eigenvalues of the correlation matrix as variance contribution ratios. The first three major components were extracted, accounting for 34.8%, 23.7% and 13.3% of the total variation, respectively. The first principal component information is derived mainly from EB, EA, NA, GRF and ICA; the second principal component information is derived primarily from XS, SA, EC, CA; the information of the third main component mainly originates from SSB, SA, CA and MPA. Fig. 5 can intuitively display the differences between samples, illustrating the distribution of samples from five enterprises. The grouped samples are subject to variation between businesses and within the same business. E, the enterprise has larger dispersivity among samples; samples D were dispersed outside the group.

4.7.2 partial least squares discriminant analysis (PLS-DA)

To better observe the differences between groups, PLS-DA with supervised mode was selected for discriminant analysis based on Principal Component Analysis (PCA). The supervised PLS-DA model has good interpretation and predictive power (R ² X＝0.980，R ² Y＝0.968，Q ² =0.698). The PLS-DA score chart shows (FIG. 3) that samples from A-E enterprises each group together, indicating a significant difference in chemical composition. Furthermore, predictive Variable Importance (VIP)>1 is used as a criterion for screening peaks of major differences between different enterprises as shown in fig. 4. RD, MPA, DS, SSB, EC, ICA and JB are index components of the major differences. These components play an important role in distinguishing between different batches of NLSCs in different enterprises and are the main differential distinguishing components. The prompt should pay attention to the feeding and quality control of rehmannia root, dwarf lilyturf tuber, thinleaf milkwort root-bark, chinese magnoliavine fruit, epimedium herb and wild jujube seed.

All samples were analyzed and judged to be unacceptable, with RD (from rehmannia), MPA (from dwarf lilyturf tuber), DS (from polygala tenuifolia), ICA (from epimedium), SSB (from shizandra berry), JB (from wild jujube seed), EC (from epimedium) failure rates of 37.50%, 33.33%, 20.83%, 62.50%, 8.33%, 70.83%, 16.67%, weight coefficients of ICA and JB of 1.0, and weight coefficients of RD, MPA, DS, SSB and EC of 0.9. Each component can reflect the feeding condition of a certain medicine flavor, and is set to be 100 minutes. Content measurement item X _i ＝A _i (content measurement result/average content). Times.W _i ，

5. Deer antler specificity detection

The LC-MS/MS identification method of the pilose antler in the Naoling capsule is established by taking the pilose antler polypeptide A (m/z 605.0- & gt 169.1 and m/z 605.0- & gt 275.1) as detection indexes.

5.1LC-ESI-MS analysis

Using a Waters ACQUITY

AB SCIEX 6500 for BEH C18 column (2.1X100 mm,1.7 μm) ⁺ The extracts were analyzed by LC-MS system (Waters corp., milford, USA). The mobile phases were solvent a (acetonitrile plus 0.1% acetic acid) and solvent B (pure water plus 0.04% acetic acid). Gradient procedure was used, flow rate 0.3ml/min, gradient elution: 0min,3% A;25min20%A. Column temperature 40 ℃, sample injection volume: 1 mul. The ESI source was operated in positive mode with ion source gas I, gas II and curtain gas set at 30, 50 and 50psi, respectively. The source temperature was 500℃and the ionization voltage was 5500v. Other parameters are default values for the instrument.

5.2 preparation of control solution

30mg of a deer antler reference (prepared by a laboratory) was taken, 10ml of a denaturation buffer and 1ml of a DTT solution were added, the mixture was shaken well, treated overnight at 80℃and taken out, cooled to room temperature, centrifuged, and 750. Mu.L of the supernatant was measured. Adding 100 μLIAA solution, reacting for 30min in dark place, mixing, and centrifuging; after desalting the sample solution, 500. Mu.L of the supernatant was measured. 25. Mu.L of bovine trypsin solution (1 mg/ml) was added, the mixture was subjected to enzymolysis at 37℃for 2 hours, and the mixture was taken out, cooled to room temperature, centrifuged, and the supernatant was taken through a 0.22 μm membrane.

5.3 preparation of sample solutions

Accurately weigh 10.0g NLSC powder and add 25mL1% ammonium bicarbonate solution. After sonication for 30 minutes at room temperature, the extract was centrifuged at 6000rpm for 5 minutes. The supernatant was then discarded. The sonication and centrifugation were repeated 4 times. The residue after centrifugation was extracted with 25mL of absolute ethanol and ultrasonic waves for 30 minutes, and the extract was centrifuged at 6000rpm for 5 minutes. Discarding supernatant, volatilizing residues, adding 10ml of denaturation buffer solution, shaking 1ml of DTT solution, standing at 80deg.C overnight, taking out, cooling to room temperature, centrifuging, collecting 750 μl of supernatant, adding 100 μl IAA solution, reacting for 30min in dark place, mixing, and centrifuging; 300 mu L of the upper layer solution is placed in an ultrafiltration centrifuge tube with the molecular weight cut-off of 3kD for centrifugation (12000 r/min, 15 min), 25 mu L of 1mg/m trypsin solution is added into the cut-off above the ultrafiltration membrane, enzymolysis is carried out for 2h at 37 ℃, the centrifuge tube is replaced for centrifugation (12000 r/min, 15 min), and the lower layer solution is taken as a sample solution.

PAPA was not detected in20 samples, and the reject ratio was 83.3%. The weight coefficient is 1. There are problems of no feeding, less feeding, adulterated feeding or unqualified feeding and serious ossification of deer antler. The method solves the bottleneck problem of animal medicine quality control technology in the traditional Chinese medicine preparation, and can provide reference for animal medicine quality control and medicine supervision.

The feeding set score of the pilose antler is 100, and the weight coefficient is 1. The score of the non-fed antler was 0.

6. Multisource information fusion

(1) The multi-source information fusion weight evaluation method comprises the following steps: (1) Detection is carried out according to standard and non-standard methods, wherein the non-standard methods comprise microscopic identification, HPLC-DAD chromatography, LC-MS/MS chromatography, principal component analysis, partial least squares discriminant analysis, deer antler specificity detection, LC-ESI-MS analysis, similarity evaluation and multi-index content measurement, and the standard method is standard number WS ₃ -B-3092-98; (2) Finding out quantifiable items with differences in the detection results of the two methods as evaluation indexes according to the detection results of the step (1); (3) Determining that the weight W of the evaluation index in the step (2) is 0.9 or 1.0, wherein the weight coefficient of the evaluation index with the failure rate lower than 50% is determined to be 1.0, and the failure rate higher than 50% is determined to be 0.9; determining the limit value of the evaluation index in the step (2) to be 80-100 respectively, and calculating the assigned value A by combining the specific conditions of each index _i The method comprises the steps of carrying out a first treatment on the surface of the (4) Calculating the score X of the evaluation item according to the assigned value and the weight coefficient in the step (3) _i ＝A _i W _i The method comprises the steps of carrying out a first treatment on the surface of the (5) Score X of the evaluation item _i The SPSSAU-selected entropy TOPSIS method was introduced to calculate the contribution coefficients S (Table 3) of the respective evaluation items, si being 0.05%, 0.09% and 9.88%, respectively、7.20％、5.91％、7.62％、1.63％、13.81％、2.85％、50.95％；

(6) According to

And sorting according to the scores from high to low so as to evaluate the relative quality. I.e. r= (0.05X ₁ +0.09X ₂ +9.88X ₃ +7.20X ₄ +5.91X ₅ +7.62X ₆ +1.63X ₇ +13.81X ₈ +2.85X ₉ +50.95X ₁₀ )/100。

The higher the R score, the better the quality.

TABLE 3 determination of the score limit, weight coefficient (W) and contribution coefficient (S) of the items

Comprehensive evaluation (table 4) shows that for 10 indexes (microscopic identification score, similarity score RD, MPA, DS, ICA, SSB, JB, EC and pilose antler feeding score), multi-source information fusion is performed, and meanwhile, 24 evaluation objects (the number of sample sizes is the number of the evaluation objects); the technology can be used for finding out the illegal production behaviors of different enterprises more effectively and providing data support for drug supervision and inspection. The NLSC weight evaluation method based on multi-source information fusion is constructed, the advantages and disadvantages of the NLSC weight evaluation method are ordered, the defect of subjective weighting is avoided, the quality of large-variety traditional Chinese medicines of different enterprises and batches is effectively evaluated, and a new path is opened up for the quality evaluation of Chinese patent medicines.

Table 4R-value calculation results for 24 samples of Naolinsu capsules

Abbreviations: DA, dehydro-deluxe; PAC, polyporus acid C; ATA pachymic acid; XS thiazine diketone glycoside; XZ, buprofezin; JA spine date seed saponin A; JB-semen Ziziphi Spinosae saponin B; MPA methyl ophiopogon root flavanone A; RD rehmannia glycoside D; GRF ginsenoside Rf; GRG ginsenoside Rg1; GRR, ginsenoside Re; GRB, ginsenoside Rb1; PX, polygala tenuifolia xanthone III; NA, neochlorogenic acid; CA, chlorogenic acid; DS3,6' -sinapiyl sucrose; EA, epimedin a; EB, epimedin B; EC, epimedin C; ICA, icariin; SA, schisandrin A; BSI baohuoside I; SSA schizandrin a; SSB schisandrin B.

Although the present disclosure describes embodiments, not every embodiment is described in terms of a single embodiment, and such description is for clarity only, and one skilled in the art will recognize that the embodiments described in the disclosure as a whole may be combined appropriately to form other embodiments that will be apparent to those skilled in the art.

Claims

1. A quality evaluation method of Chinese patent medicine based on multisource information fusion is characterized by comprising the following steps: the method comprises the following steps:

(1) Detecting according to standard and nonstandard methods, wherein the nonstandard methods comprise microscopic identification, HPLC-DAD chromatography, LC-MS/MS chromatography, chemometric analysis, deer antler specificity detection, LC-ESI-MS analysis, similarity evaluation and multi-index content measurement;

(2) Finding out quantifiable items with differences in the detection results of the two methods as evaluation indexes according to the detection results of the step (1); specifically, components with VIP (contribution value) more than 1 and pilose antler feeding items in microscopic identification, similarity evaluation and multi-index content measurement are set as evaluation indexes;

(3) Determining the weight W of the evaluation index in the step (2) to be 0.9 or 1.0, and calculating an assigned value A according to the specific conditions of each index _i ；

(4) Calculating the score X of the evaluation item according to the assigned value and the weight coefficient in the step (3) _i ＝A _i W _i ；

(6) According to

2. The quality evaluation method of the Chinese patent medicine based on multi-source information fusion as set forth in claim 1, which is characterized in that: the method is applied to the quality evaluation of the Naolingsu capsules.

3. The quality evaluation method of the Chinese patent medicine based on multi-source information fusion as claimed in claim 2, which is characterized in that: the standard method is standard number WS ₃ -B-3092-98。

4. The quality evaluation method of the Chinese patent medicine based on multi-source information fusion as claimed in claim 2, which is characterized in that: in the step (3), the weight coefficient of the evaluation index with the failure rate lower than 50% is determined to be 1.0, the failure rate higher than 50% is determined to be 0.9, and the components with the VIP (contribution value) more than 1 in the microscopic identification, similarity evaluation and multi-index content measurement and the set evaluation score limit value of the pilose antler powder feeding project are set to be 80-100.

5. The quality evaluation method of the Chinese patent medicine based on multi-source information fusion as claimed in claim 2, which is characterized in that: the microscopic identification project in the step (3) is set to be 80 in total, the weight coefficient is 0.9, and the assigned value is combined with the drug feeding condition A ₁ 60 to 80 percent; the total score of similarity evaluation is set to 100 points, and the weight coefficient is 0.9; similarity item score A ₂ =similarity result×100; seven components with contribution values greater than 1 are obtained through chemometric analysis in multi-index content measurement, and RD, MPA, DS, SSB, JB are set to 100 minutes; ICA and EC are derived from Epimedium herb and are divided into 100 points, the weight coefficients of ICA and JB are 1.0, RD, MPA, DS, SSB and ECThe weight coefficient is 0.9. Content measurement item A _i(i＝3 -9) =content measurement result/average content, the feeding of deer antler is set to a score of 100, the weight coefficient is 1, if no feeding is performed, the score is 0, namely the deer antler score A ₁₀ 0 or 100, respectively.