Donkey-hide gelatin quality control method based on near infrared spectrum

Abstract

The invention discloses a donkey-hide gelatin quality control method based on a near infrared spectrum, which establishes a quality control point associated with biological activity and reflecting quality fluctuation and a rapid detection method by comprehensively biologically evaluating the characteristics of associated biological activity and the advantage of rapid identification of a near infrared spectrum technology. And technical support is provided for realizing rapid and online detection of donkey-hide gelatin quality fluctuation and improving the quality controllability of donkey-hide gelatin products. The method specifically comprises the following steps: carrying out near infrared spectrum detection on a donkey-hide gelatin sample, combining near infrared spectrum data with biological thermal activity data to obtain a characteristic wave band highly related to biological activity as a quality control point, and establishing an effect near infrared spectrum; and finally, evaluating the comprehensive similarity of the effect near infrared spectrum to achieve the purpose of evaluating the quality of the donkey-hide gelatin product.

Description

Technical Field

[0001] The present application relates to a detection technology for donkey-hide gelatin quality, and in particular to a donkey-hide gelatin quality control method based on near-infrared spectroscopy. Background Art

[0002] Donkey hide gelatin is a solid gelatin made by boiling and concentrating the dried or fresh skin of the donkey (Equus asinm L.), an equine animal. It is sweet and mild in nature, and enters the lung, liver, and kidney meridians. It has the effects of nourishing blood and yin, moistening dryness, and stopping bleeding. It is widely used in clinical practice. In recent years, with the expansion of the market demand for donkey hide gelatin and the continuous increase in donkey hide gelatin and related products, it is difficult to unify the control of the quality of donkey hide gelatin products from different production areas and different manufacturers; the phenomenon of passing fake products off as genuine ones and inferior products as good ones has occurred frequently, which seriously affects the clinical efficacy of donkey hide gelatin and buries hidden dangers for the safe and rational use of donkey hide gelatin.

[0003] As a traditional tonic Chinese medicine, donkey-hide gelatin is widely used in clinical practice and has attracted much attention for its excellent health-care effects. However, due to the complicated production process of donkey-hide gelatin, there are many links in the production process from raw materials to finished products that are prone to cause product quality fluctuations. In addition, due to its complex ingredients and unclear active substance basis, there is a lack of effective control indicators. All of the above have caused great difficulties in the quality control of donkey-hide gelatin, making its quality issues also a focus of attention.

[0004] Among the existing quality control methods for donkey-hide gelatin, the determination of the content of four amino acids in the "Pharmacopoeia" effectively controls the consistency of high-content chemical components in donkey-hide gelatin, and the introduction of characteristic peptide identification methods has also greatly improved the efficiency of identifying the authenticity of donkey-hide gelatin.

[0005] However, as an animal medicine with complex ingredients, it is difficult to quickly and comprehensively reflect the overall quality of donkey-hide gelatin by controlling only a few indicators of chemical composition. With the development of modern science and technology, the disadvantage of simple chemical control as a double-edged sword has gradually emerged. Criminals confuse the product through biosynthesis, which increases the difficulty of quality control. The existing near-infrared spectroscopy analysis technology focuses on the identification and distinction of the authenticity, manufacturer, storage time, etc. of donkey-hide gelatin and online monitoring during the production process. However, the correlation between its detection indicators and the efficacy / activity of donkey-hide gelatin is not clear. It is difficult to reflect the integrity and specificity of the pharmacological effects of donkey-hide gelatin on organisms / even humans by relying solely on near-infrared physical and chemical detection methods for identification and detection.

[0006] Therefore, establishing a rapid quality evaluation method, especially an evaluation method related to the overall activity of donkey-hide gelatin quality, is an important issue that needs to be urgently addressed in donkey-hide gelatin quality control. Summary of the invention

[0007] In order to solve the above technical problems, the present application aims to solve the problem that the animal-based traditional Chinese medicine donkey-hide gelatin lacks quality indicators related to biological activity. By combining biological evaluation methods with physical and chemical analysis, the key substances or detection indicators that can be focused on monitoring and controlling the impact on the biological activity of donkey-hide gelatin are screened out, thereby achieving the purpose of controlling the quality of traditional Chinese medicine.

[0008] This application aims at the difficulty of quality control of animal-based traditional Chinese medicine donkey-hide gelatin. By integrating the characteristics of biological activity associated with comprehensive biological evaluation and the advantages of rapid identification using near-infrared spectroscopy technology, quality control points and rapid detection methods that are associated with biological activity and reflect quality fluctuations are established, providing technical support for rapid, on-line detection of quality fluctuations in donkey-hide gelatin and improving the quality controllability of donkey-hide gelatin products.

[0009] In the quality evaluation of traditional Chinese medicine, the biothermal activity spectrum can comprehensively evaluate a set of biological characteristic information or spectra expressed by the test drug acting on the biological system, which can better reflect the overall quality characteristics and subtle quality fluctuations of traditional Chinese medicine. It is an important method for the quality evaluation of traditional Chinese medicine, especially for traditional Chinese medicine with unclear material basis and difficult to characterize by chemical methods. This application obtains the characteristic biothermal activity spectrum of donkey-hide gelatin, and uses near-infrared spectroscopy detection, and uses the characteristic bands with high biological activity as quality control points, thereby controlling the biological activity of commercially available products and achieving the purpose of comprehensively evaluating product quality.

[0010] A method for controlling donkey-hide gelatin quality based on near infrared spectroscopy comprises the following steps:

[0011] 1) Select two or more donkey-hide gelatin samples and crush each donkey-hide gelatin sample;

[0012] 2) Collect the near infrared spectra of each donkey-hide gelatin sample to obtain the absorbance of each sample to be tested; the spectral band range is 3600cm -1 Up to 12500cm -1 ;

[0013] 3) calculating the comprehensive similarity of the absorbance of the spectral bands, and evaluating the consistency of the two or more donkey-hide gelatin samples by the comprehensive similarity.

[0014] In one embodiment provided in the present application, the donkey-hide gelatin sample in step 1) is crushed to 50 mesh to 65 mesh.

[0015] In one embodiment provided in the present application, step 2) collects near-infrared spectra of each donkey-hide gelatin sample to be tested: near-infrared spectrum data of each sample is collected more than three times, and the average value is taken as the near-infrared spectrum data of the donkey-hide gelatin sample to be tested.

[0016] In one embodiment provided in the present application, the spectral range in step 2) is 3695 cm -1 Up to 4057cm-1 、5099cm -1 Up to 5268cm -1 and 6711cm -1 Up to 6950cm -1 .

[0017] In one embodiment provided in the present application, the method for calculating the comprehensive consistency of the absorbance of the spectral band in step 3) is the angle cosine vector method.

[0018] In one embodiment provided in the present application, the conditions of the near infrared spectra of each donkey-hide gelatin sample to be tested include: solid integrating sphere-rotating spectrum acquisition mode, sample scan: 64 scans, background scan: 64 scans, resolution: 16 cm -1 , spectrum collection temperature: 25℃.

[0019] On the other hand, the present application provides an online monitoring system for donkey-hide gelatin quality, comprising: a spectrometer and a donkey-hide gelatin quality identification module, wherein the spectrometer is used to collect the near-infrared spectrum of the donkey-hide gelatin sample to obtain the absorbance value of the characteristic band;

[0020] The donkey-hide gelatin quality identification module is used to calculate the comprehensive similarity, SD value and RSD value of the absorbance of the characteristic bands of each donkey-hide gelatin sample;

[0021] The average similarity, SD value and RSD value of the absorbance of the characteristic bands of each donkey-hide gelatin sample are determined to be within the set range to determine the similarity of the donkey-hide gelatin samples.

[0022] In one embodiment provided by the present application, the higher the average similarity value (close to 1), the lower the SD value (close to 0), the lower the RSD value (close to 0), the more stable each batch of products, and the lower the product traceability re-inspection rate. The technical effects that can be achieved by the present application are:

[0023] This application uses the characteristic waveband with high correlation with biological thermal activity as the quality control point of donkey-hide gelatin, and applies it to the industrial production of donkey-hide gelatin products, so as to realize the rapid and online detection of the overall quality of donkey-hide gelatin related to the biological effect or clinical efficacy of donkey-hide gelatin, and improve the quality controllability of donkey-hide gelatin products.

[0024] After testing multiple batches of donkey-hide gelatin samples from the same manufacturer, it was shown that the donkey-hide gelatin products from the same manufacturer are relatively similar, indicating that this method can be used to investigate the biological activity quality of donkey-hide gelatin from the same manufacturer.

[0025] After testing samples of donkey-hide gelatin from different manufacturers, it was found that the similarity of the absorbance of the spectral bands of donkey-hide gelatin from different manufacturers had certain differences, indicating that this method can be used to investigate the quality of the biological activity of commercially available donkey-hide gelatin.

[0026] After testing the absorbance similarity of the spectral bands of the finished donkey-hide gelatin pieces, it was found that compared with the existing pharmacological experiments related to donkey-hide gelatin, this method has the advantages of short time, less consumables, stable model and high repeatability, and is more suitable for precious Chinese medicines such as donkey-hide gelatin.

[0027] The characteristic wavebands with high correlation with biological activity provided in the present application can more quickly and comprehensively characterize the information of biological thermal activity, thereby more truly reflecting the effects of donkey-hide gelatin on organisms and even the human body.

[0028] This application is based on similarity calculation and can evaluate the consistency of donkey-hide gelatin samples more quickly.

[0029] Other features and advantages of the present application will be described in the following description, and partly become apparent from the description, or understood by practicing the present application. The purpose and other advantages of the present application can be realized and obtained by the structures specifically pointed out in the description, claims and drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] The accompanying drawings are used to provide further understanding of the technical solution of the present application and constitute a part of the specification. Together with the embodiments of the present application, they are used to explain the technical solution of the present application and do not constitute a limitation on the technical solution of the present application.

[0031] Figure 1A The near infrared spectroscopic analysis diagram of 10 batches of donkey hide gelatin samples from 5 manufacturers in Example 1;

[0032] Figure 1B This is a near infrared spectrum principal component score graph of near infrared spectrum analysis of 10 batches of donkey-hide gelatin samples from 5 manufacturers in Example 1;

[0033] Figure 1C This is the spectrum-effect correlation analysis of near-infrared spectra of 10 batches of donkey-hide gelatin samples from 5 manufacturers in Example 1;

[0034] Figure 1D The near-infrared spectra of 10 batches of donkey-hide gelatin samples from 5 manufacturers in Example 1 are the relevant bands of t[1] and t[2] (orange is t[1], blue is t[2]).

[0035] Figure 2A , Figure 2B , Figure 2C and Figure 2D The second-order derivatives of near-infrared spectra and characteristic band second-order derivatives of 10 batches of donkey-hide gelatin samples from 5 manufacturers in Example 1;

[0036] Figure 3A , Figure 3B , Figure 3C , Figure 3D and Figure 3E The biothermal activity spectra of donkey-hide gelatin samples from 5 manufacturers;

[0037] Figure 3F This is the principal component score diagram of the effect of 10 batches of donkey-hide gelatin from 5 manufacturers on the biothermal activity of Escherichia coli;

[0038] Figure 3G for Figure 3F The first principal component t[1] in

[0039] Figure 3H for Figure 3F The second principal component t[2] in

[0040] Figure 4A , Figure 4B , Figure 4C and Figure 4D The second-order derivative of the near-infrared spectra and the second-order derivative of the characteristic band of 10 batches of donkey-hide gelatin samples from the DE manufacturer in Example 2;

[0041] Figure 5A The near infrared spectra of 10 batches of donkey-hide gelatin;

[0042] Figure 5B This is the near infrared spectrum principal component score diagram of the near infrared spectrum analysis of 10 batches of donkey-hide gelatin samples;

[0043] Figure 5C for Figure 5B The first principal component t[1] in

[0044] Figure 5D for Figure 5B The second principal component t[2] in . DETAILED DESCRIPTION

[0045] In order to make the purpose, technical solution and advantages of the present application more clear, the embodiments of the present application will be described in detail below with reference to the accompanying drawings. It should be noted that the embodiments and features in the embodiments of the present application can be combined with each other arbitrarily without conflict.

[0046] The equipment used in the embodiment is: Fourier transform near-infrared spectrometer (MPA BRUKER, GerDEny); TADEir milliwatt thermal conductivity isothermal microcalorimeter (Swedish Thermometric Company, equipped with 8 independent channels, can measure 8 samples simultaneously; detection limit is 4μW; precision is ±20μW; error is less than ±23μW; system temperature is controlled at 37°C); vertical pressure steam sterilizer (Chongqing Yamato Technology Co., Ltd.); XS205DU electronic balance (Swiss Metter Toledo Sartorious Company); SW-CT-2FD double-person single-sided clean bench (Suzhou Purification Equipment Factory); TH2-22 desktop constant temperature oscillator (Taicang Experimental Equipment Factory, Jiangsu Province), SynergH2 full-function microplate detector (BioTek Company, USA).

[0047] The detection conditions used in the embodiment are: detection at a constant temperature of 37°C.

[0048] The bacterial species used in the examples are: Escherichiacolia coli ATCC 25922 (provided by the National Institute for the Control of Pharmaceutical and Biological Products); LB broth medium (purchased from Beijing Aoboxing Biotechnology Co., Ltd.).

[0049] The analysis process in the embodiment also used the biothermodynamic data analysis software V1.0 copyright registration number: 2011SR0955405.

[0050] Raw materials used in the examples: 18 batches of donkey hide gelatin from 5 different manufacturers were purchased commercially, and detailed information is shown in Table 1.

[0051] Table 1: Sample information table

[0052] serial number factory batch number category Origin weight S1 DE 1612024 Donkey-hide gelatin slices Shandong 90g S2 DE 1612026 Donkey-hide gelatin slices Shandong 90g S3 DE 1612022 Donkey-hide gelatin slices Shandong 90g S4 DE 1612023 Donkey-hide gelatin slices Shandong 90g S5 DE 1612025 Donkey-hide gelatin slices Shandong 90g S6 DE 1612027 Donkey-hide gelatin slices Shandong 90g S7 DE 1612028 Donkey-hide gelatin slices Shandong 90g S8 DE 1612029 Donkey-hide gelatin slices Shandong 90g S9 DE 1612030 Donkey-hide gelatin slices Shandong 90g S10 DE 1612031 Donkey-hide gelatin slices Shandong 90g S11 FRT 20120501 Donkey-hide gelatin slices Henan 90g S12 FRT 20131012 Donkey-hide gelatin slices Henan 90g S13 GJT 151222 Donkey-hide gelatin slices Shandong 90g S14 GJT 160111 Donkey-hide gelatin slices Shandong 90g S15 HP 140206 Donkey-hide gelatin slices Hebei 90g S16 HP 140205 Donkey-hide gelatin slices Hebei 90g S17 TRT 201408093 Donkey-hide gelatin slices Beijing 90g S18 TRT 201506094 Donkey-hide gelatin slices Beijing 90g

[0053] In the examples of this application, microcalorimetry is involved, and the conclusion of the consistency of the donkey-hide gelatin sample obtained by near-infrared spectroscopy is verified by microcalorimetry. The preparation, conditions, and instruments used are as described above. The specific operation is as follows:

[0054] The microcalorimetric method for determining the consistency of donkey-hide gelatin samples includes the following steps:

[0055] 1. Preparation of initial bacterial solution

[0056] Accurately weigh 10.0g of LB broth medium powder and dissolve it in 500ml of distilled water, adjust the pH value to 7.0-7.2, sterilize it with high pressure steam at 121℃ for 30min, cool it to room temperature and store it in a refrigerator at 4℃ for later use. Inoculate Escherichia coli (ATCC25922) into a centrifuge tube containing 10ml of LB broth medium and shake the bacteria overnight. When the relative OD value of the bacterial solution is 0.3 using an ELISA reader (i.e., microplate reader) (wavelength = 600nm), the prepared original Escherichia coli solution is obtained.

[0057] 2. Preparation of test bacterial solution

[0058] The concentration of the bacterial solution was selected to be 1×10 5 times for mixing with donkey-hide gelatin samples. The concentration of Escherichia coli in donkey-hide gelatin samples of different concentrations was diluted 1×10 7 times the concentration.

[0059] 3. Preparation of test samples

[0060] Accurately weigh the required concentration of gelatin samples of the batch to be tested into a vial, add 10 ml of culture medium to each sample, so that the final concentration of gelatin in the liquid is 5.0 mg / ml, sterilize with high-pressure steam at 121°C for 30 min, and place in a clean bench to cool to room temperature.

[0061] 4. Detection of donkey-hide gelatin concentration

[0062] Under the condition of constant temperature of 37℃, the microcalorimetry method was used for determination, that is, the 0mg / ml donkey-hide gelatin sample model group. Take 100μl of the equally diluted Escherichia coli liquid in step 3 and add it to the newly configured 10ml test samples of different concentrations prepared in step 4, so that the concentration of the test sample bacterial liquid is consistent, seal them separately, shake them well, and place them in the microcalorimeter for detection. Record the heat generation power-time (Pt) curve of each concentration. When the curve returns to the baseline, the experiment ends.

[0063] 5. Screening of evaluation parameters:

[0064] The heat production power-time (Pt) curve obtained in step 5 was plotted using Origin9.0 software, and the following biothermodynamic parameters were extracted: F1, LHW1, RHW1, Q1, AP1 (%), LHW2, RHW2, F2, P E and EX2, their meanings are shown in Table 2.

[0065] Table 2 Information on main parameters of biothermodynamics

[0066]

[0067]

[0068] In order to ensure the stability of the Escherichia coli model and the applicability of the detection dose after administration of donkey-hide gelatin, the present application conducted a methodological investigation on the effect of donkey-hide gelatin solution at a concentration of 5.0 mg / ml on the Escherichia coli model, and repeated experiments were performed according to steps 5 and 6 to obtain 10 evaluation parameters.

[0069] 9. Model construction

[0070] The 10 parameters screened in step 6) are used as a fixed parameter combination to obtain the calculation formula for the principal components t[1] and t[2] values ​​of the evaluation index based on the dose-effect relationship, that is, the principal component analysis prediction calculation formula.

[0071]

[0072]

[0073] Y={0.91,0.89,0.92,1.10,0.74,0.94,0.84,0.87,0.82,1.22}

[0074]

[0075] P1={0.36,0.38,0.34,-0.29,0.58,0.18,0.20,0.03,0.26,-0.20}

[0076] P2={-0.16,-0.15,-0.14,0.08,-0.22,0.20,0.41,0.56,0.25,-0.56}

[0077] X={F1,LHW1,RHW1,Q1,AP1(%),LHW2,RHW2,F2,P E ,EX2}

[0078] X is the normalized biothermodynamic parameter, and the normalized processing is the ratio of the biothermodynamic parameter of the donkey-hide gelatin sample extracted by Origin9.0 software to the biothermodynamic parameter of the model group with the donkey-hide gelatin sample concentration of 0 mg / ml.

[0079] Since the principal components t[1] and t[2] can comprehensively evaluate the information in the biothermal activity spectrum, they are used as characteristic indicators to evaluate the consistency of donkey-hide gelatin quality. The smaller the SD value of the evaluation indicators t[1] and t[2] in multiple batches of tests, the better the consistency.

[0080] Example 1

[0081] Ten batches of donkey-hide gelatin samples (S1, S2, S11-S18) were collected from 5 manufacturers and each manufacturer, and the donkey-hide gelatin samples were crushed to 50-65 mesh;

[0082] The near infrared spectrum data and characteristic band spectrum data of each donkey-hide gelatin sample were collected three times, and the average value of the three data was taken. Figure 1A .

[0083] The principal component analysis software (Simca-p14 (Umetrics, MKS Instruments Inc, Sweden) was used to perform principal component analysis on the near-infrared spectral data of 40 batches of donkey-hide gelatin samples collected in all bands. Figure 2B The HP of the middle manufacturer deviated far away, while the manufacturers DE, FRT, GJT, and TRT were relatively clustered, indicating that the chemical compositions were relatively similar, and the FRT, which had a significantly different effect on E. coli than other manufacturers, could be effectively distinguished.

[0084] The second-order derivative analysis of the near-infrared spectrum data and characteristic band data of 10 batches of donkey-hide gelatin products from five manufacturers was performed. The second-order derivative of the near-infrared spectrum and the second-order derivative of the characteristic band are as follows: FIG. 3A to FIG. 3E As shown in the figure, it can be seen that the characteristic bands are relatively similar, and the calculated characteristic band is 3695cm -1 Up to 4057cm -1 、5099cm -1 Up to 5268cm -1 and 6711cm -1 Up to 6950cm -1 The results showed that the average similarity of the characteristic bands of 10 batches of donkey-hide gelatin from 5 manufacturers was 0.871, SD≈0.103, and RSD≈11.84%;

[0085] Table 3 Similarity of characteristic bands of 10 batches of donkey-hide gelatin products from five manufacturers

[0086]

[0087] Verification of Example 1:

[0088] The bioactivity data of 10 batches of donkey-hide gelatin samples from the above five manufacturers were collected by microcalorimetry; the consistency of the donkey-hide gelatin samples was verified by consistency analysis based on biothermal activity.

[0089] 5mg·ml -1 The effect of donkey-hide gelatin mass concentration on the growth and metabolism of Escherichia coli was tested and analyzed. The 10 batches of donkey-hide gelatin samples from 5 manufacturers had different effects on the growth and metabolism of Escherichia coli and the spectra were significantly different ( FIG. 3A to FIG. 3F ).by Figure 3F The dose-effect relationship reflected in the model was used to predict the effects of different manufacturers' donkey-hide gelatin on the growth and metabolism of Escherichia coli. The results showed that there were certain differences in the direction of action of donkey-hide gelatin products from different manufacturers, and each manufacturer had a certain emphasis on the first and second principal components ( Figure 3F , Figure 3G and Figure 3H ).

[0090] Example 1 was verified; microcalorimetry was used to calculate that t[1] and t[2] of 10 batches of samples are shown in the following table:

[0091] Table 4. t[1] and t[2] values ​​obtained by microcalorimetry for 10 batches of samples from five manufacturers

[0092] batch t[1] t[2] 1612024 0.0967017 -0.311341 1612026 0.403303 0.169069 20120501 -2.29755 -1.81193 20131012 -1.60313 -1.10635 1512225 0.951962 -0.308864 160111 1.25941 -0.421557 140206 -1.17167 0.239562 140205 -0.732325 0.701271 201408093 1.65268 1.09755 201506094 1.88254 1.53272

[0093] Among them, the SD value of t[1] is ±1.441, and the SD value of t[2] is ±1.000, indicating that there are differences between manufacturers. Compared with the conclusion drawn by near infrared spectroscopy, the method described in this application can better distinguish the differences between manufacturers.

[0094] Example 2

[0095] Ten batches of donkey-hide gelatin samples (S1-S10) from the same manufacturer (manufacturer A) with the same production process and similar production time were collected, and the donkey-hide gelatin samples were crushed to 50-65 mesh;

[0096] The near infrared spectrum data and characteristic band spectrum data of each donkey-hide gelatin sample were collected three times, and the average value of the three data was taken. Figure 5A .

[0097] After principal component analysis ( Figure 5B ), the 10 batches of donkey-hide gelatin samples from manufacturer DE were relatively clustered, indicating that the chemical compositions were relatively similar.

[0098] Collect its near-infrared spectrum data and characteristic band 3695cm -1 Up to 4057cm -1 、5099cm -1 Up to 5268cm -1 and 6711cm -1 Up to 6950cm -1 The near infrared spectrum data and characteristic band spectrum data were collected three times for each donkey-hide gelatin sample, and the average of the three data was taken. The second-order derivative analysis of the near infrared spectrum data and characteristic band data was performed. The second-order derivative of the near infrared spectrum and the second-order derivative of the characteristic band are shown in FIG. 4A to FIG. 4D As shown:

[0099] It can be seen from the figure that the characteristic bands of the 10 batches are relatively similar, and the calculated characteristic band is 3695cm -1 Up to 4057cm -1 、5099cm -1 Up to 5268cm -1 and 6711cm -1 Up to 6950cm -1 The results showed that the average similarity of the characteristic bands of 10 batches of donkey-hide gelatin from DE manufacturer was 0.843, SD≈0.062, and RSD≈7.31%.

[0100] It can be seen that some of the 10 batches of DE manufacturer have large fluctuations. From the perspective of product consistency, their consistency needs to be improved.

[0101] Table 5 Similarity of characteristic bands of 10 batches of donkey-hide gelatin products from DE manufacturer

[0102]

[0103] Verification of Example 2:

[0104] The bioactivity data of 10 batches of donkey-hide gelatin samples from the above-mentioned DE manufacturer were collected by microcalorimetry; the consistency of the donkey-hide gelatin samples was verified by consistency analysis based on biothermal activity.

[0105] 5mg·ml -1 The effect of donkey-hide gelatin mass concentration on the growth and metabolism of E. coli was tested and analyzed. The effects of 10 batches of donkey-hide gelatin samples from DE manufacturer on the growth and metabolism of E. coli were relatively small. The principal component prediction analysis of the near-infrared spectra and biothermal activity fingerprints of 10 batches of donkey-hide gelatin is shown in Figure 2. FIG. 5A to FIG. 5D shown.

[0106] Example 2 was verified; microcalorimetry was used to calculate that t[1] and t[2] of 10 batches of samples are shown in the following table:

[0107] Table 6. t[1] and t[2] values ​​obtained by microcalorimetry for 10 batches of samples from DE manufacturer

[0108] batch t[1] t[2] 1612022 1.42272 0.442685 1612023 0.31496 0.316712 1612024 0.0967017 -0.311341 1612025 0.228359 0.0732138 1612026 0.403303 0.169069 1612027 0.634865 0.534258 1612028 0.0915963 -0.196341 1612029 1.39913 0.367804 1612030 1.13942 0.471942 1612031 0.909236 0.367219

[0109] Among them, the SD value of t[1] is ±0.5198, and the SD value of t[2] is ±0.2875, indicating that the differences among the 10 batches are small and the quality is stable, which is consistent with the conclusion drawn by near-infrared spectroscopy.

[0110] Based on Examples 1 and 2, the functional groups and substance types of the three characteristic bands were confirmed. Analysis revealed that the products that caused the largest fluctuations in biological thermal activity were mostly products of protein fragmentation, such as carboxylic acids and amides.

[0111] It can be seen that there is a band in the near-infrared spectral data that is highly correlated with the effect of donkey-hide gelatin on the growth and metabolism of intestinal flora: 3695cm -1 Up to 4011cm -1 5245cm -1 Up to 5268cm -1 、6711cm -1 Up to 6950cm -1 , which can be used as the characteristic bands for evaluating the quality consistency of donkey-hide gelatin as the effect near-infrared spectrum. When the effect near-infrared spectrum is applied to the quality evaluation of donkey-hide gelatin, the above three near-infrared characteristic bands have a good correlation with the biological activity and can reflect the changes in the overall biological activity of donkey-hide gelatin. And it can more comprehensively evaluate the consistency and consistency of the quality of different donkey-hide gelatin.

[0112] In addition, SPSS software was used to perform Pearson correlation analysis on the near-infrared spectrum and biothermal activity fingerprint spectrum data. Correlation analysis was performed on the t[1] and t[2] principal component scores of 10 batches of DE manufacturers and the near-infrared spectrum data. The results showed that the correlation coefficients of the above three bands with the t[1] principal component were ≥0.7, and the correlation coefficients with the t[2] principal component were ≥0.5.

[0113] In summary, this application can evaluate the quality consistency of donkey-hide gelatin slices by measuring the three characteristic bands of different batches or different manufacturers. This application is applied to the industrial production of donkey-hide gelatin products, and can realize the rapid and online detection of the overall quality of donkey-hide gelatin related to the biological effect or clinical efficacy of donkey-hide gelatin, thereby improving the quality controllability of donkey-hide gelatin products. For example, by using near-infrared spectroscopy to analyze the authenticity, manufacturer, storage time, etc. of the product, the biological activity of commercially available products can be predicted, and ultimately the purpose of comprehensively evaluating product quality can be achieved; and for donkey-hide gelatin products from the same manufacturer, this method can be applied to the quality control of the production process, by focusing on the characteristic bands that are highly correlated with the biological activity of the manufacturer's products, the product quality can be improved, and the consistency of the production products can be ensured.

Claims

1. A method for quality control of donkey-hide gelatin based on near infrared spectroscopy, comprising the following steps: 1) Select two or more donkey-hide gelatin samples and crush each donkey-hide gelatin sample; 2) Collect the near infrared spectra of each donkey-hide gelatin sample to obtain the absorbance of each sample to be tested; the spectral band range is 3600cm -1 Up to 12500cm -1 ; 3) calculating the comprehensive similarity of the absorbance of the spectral bands, and evaluating the consistency of the two or more donkey-hide gelatin samples by the comprehensive similarity.

2. The method for controlling donkey-hide gelatin quality based on near infrared spectroscopy according to claim 1, wherein: The donkey-hide gelatin sample in step 1) is crushed to 50 to 65 meshes.

3. The method for controlling donkey-hide gelatin quality based on near infrared spectroscopy according to claim 1, wherein: Step 2) Collecting near-infrared spectra of each donkey-hide gelatin sample to be tested: collecting near-infrared spectrum data of each sample more than three times, and taking the average value as the near-infrared spectrum data of the donkey-hide gelatin sample to be tested.

4. The method for controlling donkey-hide gelatin quality based on near infrared spectroscopy according to any one of claims 1 to 3, wherein: The wavelength range of the spectrum in step 2) is selected from 3695cm -1 Up to 4057cm -1 、5099cm -1 Up to 5268cm -1 and 6711cm -1 Up to 6950cm -1 .

5. The method for controlling donkey-hide gelatin quality based on near infrared spectroscopy according to any one of claims 1 to 3, wherein: The method for calculating the comprehensive consistency of the absorbance of the spectral band in step 3) is the angle cosine vector method.

6. The method for controlling donkey-hide gelatin quality based on near infrared spectroscopy according to any one of claims 1 to 3, wherein: The conditions of the near infrared spectra of each donkey-hide gelatin sample to be tested include: solid integrating sphere-rotating spectrum acquisition mode, sample scan: 64 scans, background scan: 64 scans, resolution: 16 cm -1 , spectrum collection temperature: 25℃.

7. An online monitoring system for donkey-hide gelatin quality, comprising: A spectrometer and a donkey-hide gelatin quality identification module, wherein the spectrometer is used to collect the near-infrared spectrum of the donkey-hide gelatin sample and obtain the absorbance value of the characteristic band; The donkey-hide gelatin quality identification module is used to calculate the comprehensive similarity, SD value and RSD value of the absorbance of the characteristic bands of each donkey-hide gelatin sample; The average similarity, SD value and RSD value of the absorbance of the characteristic bands of each donkey-hide gelatin sample are determined to be within the set range to determine the similarity of the donkey-hide gelatin samples.

8. The online monitoring system for donkey-hide gelatin quality according to claim 7, wherein: The higher the average similarity value is (close to 1), the lower the SD value is (close to 0), and the lower the RSD value is (close to 0), the more stable the batches of products are, and the lower the product traceability re-inspection rate is.

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