Intelligent monitoring and analysis method for polypeptide extract preparation process

Abstract

The invention relates to the field of polypeptide extract preparation process monitoring, and specifically discloses an intelligent monitoring and analysis method for polypeptide extract preparation process. The invention monitors the process parameters of enzymatic hydrolysis, separation, purification, concentration and drying in the preparation process of spleen polypeptide extract, determines whether the requirements are met and performs adaptive adjustment, avoids the spleen polypeptide extract preparation process parameter deviation being too large to cause the spleen polypeptide structure to change, so that the spleen polypeptide extract maintains its natural active conformation, better exerts its due efficacy, and is conducive to the optimization and improvement of the spleen polypeptide extract preparation process; the polypeptide sequences contained in the spleen polypeptide extract and the spleen polypeptide injection are detected by mass spectrometry analysis technology, the components and functions of the spleen polypeptide extract and the spleen polypeptide injection are compared, and then the quality of the prepared spleen polypeptide extract is evaluated, thereby improving the spleen polypeptide extract quality evaluation system and improving the reliability and accuracy of the evaluation result.

Description

Technical Field

[0001] The invention relates to the field of polypeptide extract preparation process monitoring, and particularly relates to an intelligent monitoring and analysis method for polypeptide extract preparation process. Background Art

[0002] Spleen polypeptide extract is the precursor product of spleen polypeptide injection. It is a sterile aqueous solution of polypeptides with a molecular weight less than 6000 Daltons, free amino acids, nucleic acids, and total sugars made from healthy calf spleen extract. Spleen polypeptide extract can be used for primary and secondary cellular immunodeficiency diseases, respiratory tract and lung infections, and is used as an auxiliary in the treatment of leukopenia caused by radiotherapy and chemotherapy, leukemia, aplastic anemia, lymphoma and other malignant tumors, and to improve the malignant transformation of tumor patients and improve the physical weakness of postoperative or critically ill patients.

[0003] The activity of spleen polypeptide is closely related to its structure. If the temperature, pH value, reaction time and other factors in the preparation process of spleen polypeptide extract are not properly controlled, it may cause the spleen polypeptide structure to change and lose its natural active conformation, and thus fail to exert its due efficacy. At the same time, if impurities are introduced during the preparation process of spleen polypeptide extract, the purity of the final spleen polypeptide extract cannot be guaranteed. Therefore, it is of great significance to monitor and analyze the preparation process of spleen polypeptide extract.

[0004] However, the existing spleen polypeptide extract preparation process monitoring and analysis methods still have some limitations and shortcomings in practical applications.

[0005] First, the existing methods do not involve the monitoring and adaptive adjustment of the process parameters of each process in the preparation of spleen polypeptide extracts, such as enzymatic hydrolysis, separation, purification, concentration and drying. If the deviation of the process parameters of the spleen polypeptide extract preparation is too large, it will cause the polypeptide structure of the spleen polypeptide extract to change, and it will not be able to maintain its natural active conformation, and thus it will not be able to play its due role.

[0006] On the second aspect, the existing methods have some deficiencies in the detection of the finished product of the prepared spleen polypeptide extract: for example, the existing Chinese patent with the publication number CN102680602A discloses a method for determining the characteristic spectrum of polypeptide extracts, which includes: (1) taking no less than 3 batches of polypeptide extracts, adding water to make a suitable concentration, accurately taking 1µl and injecting it into a liquid chromatograph, testing according to certain chromatographic conditions, and importing the generated spectrum into the similarity evaluation system of the chromatographic characteristic spectrum of traditional Chinese medicine to obtain a standard characteristic spectrum; (2) taking the reference substance and the test substance and adding water to make a solution of suitable concentration, accurately taking 1µl of each, respectively injecting it into the liquid chromatograph, measuring according to certain chromatographic conditions, and recording the characteristic spectrum of the test substance; (3) comparing the similarity between the characteristic spectrum of the test substance and the standard characteristic spectrum. The method of the invention is used for the characteristic spectrum determination of polypeptide extracts, which overcomes the shortcomings of the traditional method of using ninhydrin color reaction analysis to control the quality of multi-component biochemical drugs, such as the inability to distinguish peptides from different sources and the inability to accurately reflect the quality status of existing multi-component biochemical drugs.

[0007] The above patent reflects the quality status of the polypeptide extract by comparing the similarity between the characteristic spectrum of the polypeptide extract and the standard characteristic spectrum, rather than detecting the polypeptide sequence contained in the polypeptide extract and the final product of the polypeptide extract, i.e., the polypeptide injection, through mass spectrometry analysis technology to identify the protein and polypeptide results of the polypeptide extract and evaluate the composition and function of the polypeptide extract. As a result, the existing method for evaluating the quality of polypeptide extracts is not perfect, reliable and accurate enough. Summary of the invention

[0008] In response to the above problems, the present invention proposes an intelligent monitoring and analysis method for a polypeptide extract preparation process. The specific technical scheme is as follows: An intelligent monitoring and analysis method for a polypeptide extract preparation process comprises the following steps: Step 1, regulation of enzymatic hydrolysis process parameters for polypeptide extract preparation: obtaining the temperature, duration, pH value and enzyme dosage of the enzymatic hydrolysis during the preparation of the spleen polypeptide extract, determining whether the enzymatic hydrolysis process parameters during the preparation of the spleen polypeptide extract meet the requirements, and performing adaptive adjustments.

[0009] Step 2: Control of process parameters for separation of polypeptide extract preparation: Obtain the centrifugal speed and centrifugal time during the separation process of the spleen polypeptide extract preparation, determine whether the process parameters for separation during the separation process of the spleen polypeptide extract preparation meet the requirements, and perform adaptive adjustments.

[0010] Step 3: Control of process parameters for preparation and purification of polypeptide extracts: Obtain the ionic strength and pH value of the purified eluent during the preparation of spleen polypeptide extracts, determine whether the purification process parameters during the preparation of spleen polypeptide extracts meet the requirements, and perform adaptive adjustments.

[0011] Step 4: Control of process parameters for preparation and concentration of polypeptide extracts: Obtain the concentration temperature and vacuum degree during the preparation of spleen polypeptide extracts, determine whether the concentration process parameters during the preparation of spleen polypeptide extracts meet the requirements, and perform adaptive adjustments.

[0012] Step 5. Control of drying process parameters for polypeptide extract preparation: Obtain the drying inlet and outlet air temperatures, spray speed and drying time during the preparation of spleen polypeptide extract, determine whether the drying process parameters during the preparation of spleen polypeptide extract meet the requirements, and perform adaptive adjustments.

[0013] Step 6. Quality inspection and analysis of the finished product of polypeptide extract preparation: The mass spectrometry peptide data of the prepared spleen polypeptide extract and spleen polypeptide injection samples are obtained through mass spectrometry detection technology, and the peak time and source protein type of each high-frequency peptide of the spleen polypeptide extract and spleen polypeptide injection samples are further obtained, and the quality evaluation index of the prepared spleen polypeptide extract is analyzed and feedback is provided.

[0014] Compared with the prior art, the intelligent monitoring and analysis method for the preparation process of a polypeptide extract described in the present invention has the following beneficial effects: 1. The present invention monitors the process parameters of enzymatic hydrolysis, separation, purification, concentration and drying during the preparation process of spleen polypeptide extracts, determines whether they meet the requirements and performs adaptive adjustments, thereby avoiding excessive deviations in the process parameters for the preparation of spleen polypeptide extracts that lead to changes in the structure of spleen polypeptides, thereby allowing the spleen polypeptide extracts to maintain their natural active conformation and better exert their due efficacy. At the same time, it is beneficial to the optimization and improvement of the preparation process of spleen polypeptide extracts and ensures product quality.

[0015] 2. The present invention detects the polypeptide sequences contained in the spleen polypeptide extract and the spleen polypeptide injection by mass spectrometry analysis technology, identifies the key proteins and polypeptide results of the spleen polypeptide extract and the spleen polypeptide injection, further compares the components and functions of the spleen polypeptide extract and the spleen polypeptide injection, and then evaluates the quality of the prepared spleen polypeptide extract, thereby improving the quality evaluation system of the spleen polypeptide extract and improving the reliability and accuracy of the evaluation results. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings required for describing the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other accompanying drawings can be obtained based on these accompanying drawings without paying creative work.

[0017] Figure 1 It is a schematic diagram of the method flow of the present invention.

[0018] Figure 2 The present invention is a monitoring structure block diagram of the spleen polypeptide extract preparation process.

[0019] Figure 3 The present invention is a structural block diagram of the quality detection of the spleen polypeptide extract finished product. DETAILED DESCRIPTION

[0020] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0021] See also Figure 1 , Figure 2 and Figure 3 As shown, the present invention provides an intelligent monitoring and analysis method for a polypeptide extract preparation process, comprising the following steps: Step 1, regulating the enzymatic hydrolysis process parameters for polypeptide extract preparation: obtaining the temperature, duration, pH value and enzyme dosage of the enzymatic hydrolysis during the preparation of the spleen polypeptide extract, determining whether the enzymatic hydrolysis process parameters during the preparation of the spleen polypeptide extract meet the requirements, and performing adaptive adjustments.

[0022] As a preferred embodiment, the specific analysis process of step 1 includes: monitoring the temperature, duration, pH value and enzyme dosage of the enzymatic hydrolysis during the preparation of the spleen polypeptide extract, which are respectively recorded as .

[0023] The reference temperature, reference time, reference pH value and reference enzyme dosage of the enzymatic hydrolysis process in the preparation process of the spleen polypeptide extract stored in the extraction database are recorded as .

[0024] By analyzing the formula Obtain the compliance coefficient of enzymatic hydrolysis process parameters in the preparation of spleen polypeptide extract ,in Indicates the correction factor of the preset enzymatic hydrolysis process parameters compliance coefficient, They represent the deviation thresholds of the preset enzymatic hydrolysis temperature, duration, pH value and enzyme dosage, respectively. They represent the preset enzymatic hydrolysis temperature, duration, pH value and enzyme dosage weights respectively. .

[0025] It should be noted that appropriate enzymes are selected according to the characteristics of the raw materials of the spleen polypeptide extract and the structure and properties of the spleen polypeptide, among which commonly used enzymes include but are not limited to: pepsin, trypsin, papain, alkaline protease, neutral protease, etc. Different enzymes have different cleavage sites and action conditions, which in turn affect the yield and quality of the spleen polypeptide extract.

[0026] It should be noted that different enzymes have their optimum temperature ranges, generally between 30° C. and 60° C. In a specific embodiment, the optimum temperature of trypsin is 37° C., and the optimum temperature of papain is 50° C. and 60° C. Too high a temperature will inactivate the enzyme, while too low a temperature will reduce the efficiency of enzymolysis.

[0027] It should be noted that the enzymatic hydrolysis time usually ranges from several hours to dozens of hours, which depends on the properties of the spleen polypeptide extract raw material, the type and amount of enzyme, and the requirements of the spleen polypeptide.

[0028] It should be noted that different enzymes have their optimum pH ranges. In a specific embodiment, pepsin has the highest activity at a pH of 2-3, and alkaline protease has the highest activity at a pH of 8-10.

[0029] It should be noted that too little enzyme dosage will result in incomplete enzymolysis, while too much enzyme dosage may increase costs and may cause unnecessary side reactions.

[0030] As a preferred solution, the specific analysis process of step one also includes: D1: comparing the compliance coefficient of the enzymatic hydrolysis process parameters in the preparation process of the spleen polypeptide extract with a preset enzymatic hydrolysis process parameter compliance coefficient threshold; if the compliance coefficient of the enzymatic hydrolysis process parameters in the preparation process of the spleen polypeptide extract is less than the preset enzymatic hydrolysis process parameter compliance coefficient threshold, the enzymatic hydrolysis process parameters in the preparation process of the spleen polypeptide extract do not meet the requirements, and D2 is executed.

[0031] D2: Compare the difference between the enzymatic hydrolysis temperature and the reference temperature during the preparation of the spleen polypeptide extract with the preset enzymatic hydrolysis temperature deviation threshold. If the difference between the enzymatic hydrolysis temperature and the reference temperature during the preparation of the spleen polypeptide extract is greater than the preset enzymatic hydrolysis temperature deviation threshold, the enzymatic hydrolysis temperature during the preparation of the spleen polypeptide extract needs to be adjusted, and further obtain the adjustment direction and adjustment amount of the enzymatic hydrolysis temperature during the preparation of the spleen polypeptide extract.

[0032] It should be noted that the specific method for obtaining the adjustment direction and adjustment amount of the enzymatic hydrolysis temperature during the preparation of the spleen polypeptide extract is as follows: the absolute value of the difference between the enzymatic hydrolysis temperature during the preparation of the spleen polypeptide extract and the reference temperature is used as the adjustment amount of the enzymatic hydrolysis temperature during the preparation of the spleen polypeptide extract.

[0033] The temperature of enzymatic hydrolysis during the preparation of the spleen polypeptide extract is compared with the reference temperature. If the temperature of enzymatic hydrolysis during the preparation of the spleen polypeptide extract is greater than the reference temperature, the direction of adjusting the temperature of enzymatic hydrolysis during the preparation of the spleen polypeptide extract is to decrease. If the temperature of enzymatic hydrolysis during the preparation of the spleen polypeptide extract is less than the reference temperature, the direction of adjusting the temperature of enzymatic hydrolysis during the preparation of the spleen polypeptide extract is to increase.

[0034] D3: Similarly, according to the analysis method of D2, determine whether the enzymatic hydrolysis time, pH value and enzyme dosage in the preparation process of spleen polypeptide extract need to be adjusted and analyze the adjustment direction and amount.

[0035] Step 2: Control of process parameters for separation of polypeptide extract preparation: Obtain the centrifugal speed and centrifugal time during the separation process of the spleen polypeptide extract preparation, determine whether the process parameters for separation during the separation process of the spleen polypeptide extract preparation meet the requirements, and perform adaptive adjustments.

[0036] As a preferred embodiment, the specific analysis process of step 2 is: monitoring the centrifugal speed and centrifugal time of separation during the preparation of the spleen polypeptide extract, and extracting the reference centrifugal speed and reference centrifugal time of separation during the preparation of the spleen polypeptide extract stored in the database, and comparing them to obtain the difference between the centrifugal speed and centrifugal time of separation during the preparation of the spleen polypeptide extract, and substituting it into the relationship function between the preset difference between the centrifugal speed and centrifugal time of separation and the compliance coefficient of the separation process parameters, to obtain the compliance coefficient of the separation process parameters during the preparation of the spleen polypeptide extract.

[0037] According to the compliance coefficient of the separation process parameters in the preparation process of the spleen polypeptide extract, it is judged whether the separation process parameters in the preparation process of the spleen polypeptide extract meet the requirements. If they do not meet the requirements, it is judged whether the centrifugal speed and centrifugal time of the separation in the preparation process of the spleen polypeptide extract need to be adjusted and the adjustment direction and adjustment amount are obtained.

[0038] It should be noted that the adaptive adjustment process of the separation process parameters in the preparation process of the spleen polypeptide extract is: comparing the compliance coefficient of the separation process parameters in the preparation process of the spleen polypeptide extract with the preset threshold value of the separation process parameter compliance coefficient. If the compliance coefficient of the separation process parameters in the preparation process of the spleen polypeptide extract is less than the preset threshold value of the separation process parameter compliance coefficient, the separation process parameters in the preparation process of the spleen polypeptide extract do not meet the requirements.

[0039] According to the centrifugal speed, centrifugal time and reference centrifugal speed and reference centrifugal time during the preparation of the spleen polypeptide extract, it is determined whether the centrifugal speed and centrifugal time during the preparation of the spleen polypeptide extract need to be adjusted and the adjustment direction and adjustment amount are obtained.

[0040] It should be noted that separation refers to centrifuging the mixture after enzymatic hydrolysis to remove unreacted raw materials, enzymes and other insoluble impurities. In a specific embodiment, the centrifugal speed is 3000-10000 rpm and the centrifugal time is 10-30 minutes.

[0041] It should be noted that the method for determining whether the centrifugal speed and centrifugal time for separation during the preparation of spleen polypeptide extracts need to be adjusted and obtaining the adjustment direction and amount is based on the same principle as the method for determining whether the temperature of enzymatic hydrolysis during the preparation of spleen polypeptide extracts needs to be adjusted and analyzing the adjustment direction and amount.

[0042] Step 3: Control of process parameters for preparation and purification of polypeptide extracts: Obtain the ionic strength and pH value of the purified eluent during the preparation of spleen polypeptide extracts, determine whether the purification process parameters during the preparation of spleen polypeptide extracts meet the requirements, and perform adaptive adjustments.

[0043] As a preferred embodiment, the specific analysis process of step three is: monitoring the ionic strength and pH value of the purified eluent during the preparation of the spleen polypeptide extract, and comparing them with the ionic strength and pH value of the reference eluent purified during the preparation of the spleen polypeptide extract stored in the database, and analyzing the compliance coefficient of the purification process parameters during the preparation of the spleen polypeptide extract.

[0044] It should be noted that the compliance coefficient of the purification process parameters in the preparation process of the spleen polypeptide extract is analyzed by monitoring the ionic strength and pH value of the purified eluent in the preparation process of the spleen polypeptide extract, and comparing them with the reference ionic strength and pH value of the purified eluent in the preparation process of the spleen polypeptide extract stored in the database to obtain the difference between the ionic strength and pH value of the purified eluent in the preparation process of the spleen polypeptide extract, and substituting it into the relationship function between the preset difference between the ionic strength and pH value of the purified eluent and the compliance coefficient of the purification process parameters to obtain the compliance coefficient of the purification process parameters in the preparation process of the spleen polypeptide extract.

[0045] According to the compliance coefficient of the purification process parameters in the preparation process of the spleen polypeptide extract, it is judged whether the purification process parameters in the preparation process of the spleen polypeptide extract meet the requirements. If they do not meet the requirements, it is judged whether the ionic strength and pH value of the purified eluent in the preparation process of the spleen polypeptide extract need to be adjusted and the adjustment direction and adjustment amount are obtained.

[0046] It should be noted that the adaptive adjustment process of the purification process parameters during the preparation of the spleen polypeptide extract is: comparing the compliance coefficient of the purification process parameters during the preparation of the spleen polypeptide extract with the preset threshold value of the purification process parameter compliance coefficient. If the compliance coefficient of the purification process parameters during the preparation of the spleen polypeptide extract is less than the preset threshold value of the purification process parameter compliance coefficient, the purification process parameters during the preparation of the spleen polypeptide extract do not meet the requirements.

[0047] According to the purified eluent ionic strength, eluent pH value and the reference eluent ionic strength, reference eluent pH value in the preparation process of the spleen polypeptide extract, it is determined whether the purified eluent ionic strength and eluent pH value in the preparation process of the spleen polypeptide extract need to be adjusted and the adjustment direction and adjustment amount are obtained.

[0048] It should be noted that the method for determining whether the ionic strength and pH value of the purified eluent during the preparation of the spleen polypeptide extract need to be adjusted and obtaining the adjustment direction and amount is the same as the method for determining whether the temperature of the enzymatic hydrolysis during the preparation of the spleen polypeptide extract needs to be adjusted and analyzing the adjustment direction and amount.

[0049] It should be noted that the purification in the preparation process of the spleen polypeptide extract of the present invention adopts a chromatography purification method of ion exchange chromatography, that is, separation is performed according to the different charge properties of the spleen polypeptide.

[0050] It should be noted that the ionic strength of the purified eluent during the preparation of the spleen polypeptide extract is adjusted according to the concentration of the sodium chloride solution, and the pH value of the eluent is adjusted according to the isoelectric point of the spleen polypeptide and the characteristics of the ion exchanger.

[0051] In another specific embodiment, the spleen polypeptide extract can also be purified by gel filtration chromatography or reverse phase high performance liquid chromatography and other chromatography purification methods during the preparation process.

[0052] Step 4: Control of process parameters for preparation and concentration of polypeptide extracts: Obtain the concentration temperature and vacuum degree during the preparation of spleen polypeptide extracts, determine whether the concentration process parameters during the preparation of spleen polypeptide extracts meet the requirements, and perform adaptive adjustments.

[0053] As a preferred solution, the specific analysis process of step 4 is: monitoring the concentration temperature and vacuum degree during the preparation of the spleen polypeptide extract, and comparing them with the reference temperature and reference vacuum degree during the preparation of the spleen polypeptide extract stored in the database, respectively, to analyze the compliance coefficient of the concentration process parameters during the preparation of the spleen polypeptide extract.

[0054] It should be noted that the compliance coefficient of the concentration process parameters in the preparation process of the spleen polypeptide extract is analyzed by substituting the difference in the concentration temperature and vacuum degree in the preparation process of the spleen polypeptide extract into the relationship function between the preset difference in the concentration temperature and vacuum degree and the compliance coefficient of the concentration process parameters to obtain the compliance coefficient of the concentration process parameters in the preparation process of the spleen polypeptide extract.

[0055] According to the compliance coefficient of the concentration process parameters in the preparation process of the spleen polypeptide extract, it is judged whether the concentration process parameters in the preparation process of the spleen polypeptide extract meet the requirements. If they do not meet the requirements, it is judged whether the concentration temperature and vacuum degree in the preparation process of the spleen polypeptide extract need to be adjusted and the adjustment direction and adjustment amount are obtained.

[0056] It should be noted that the adaptive adjustment process of the concentration process parameters in the preparation process of the spleen polypeptide extract is: comparing the compliance coefficient of the concentration process parameters in the preparation process of the spleen polypeptide extract with the preset threshold value of the concentration process parameter compliance coefficient. If the compliance coefficient of the concentration process parameters in the preparation process of the spleen polypeptide extract is less than the preset threshold value of the concentration process parameter compliance coefficient, the concentration process parameters in the preparation process of the spleen polypeptide extract do not meet the requirements.

[0057] According to the concentrated temperature, vacuum degree and reference temperature and reference vacuum degree in the preparation process of spleen polypeptide extract, it is judged whether the concentrated temperature and vacuum degree in the preparation process of spleen polypeptide extract need to be adjusted and the adjustment direction and adjustment amount are obtained.

[0058] It should be noted that the method for determining whether the concentration temperature and vacuum degree need to be adjusted during the preparation of the spleen polypeptide extract and obtaining the adjustment direction and adjustment amount is the same as the method for determining whether the enzymatic hydrolysis temperature needs to be adjusted during the preparation of the spleen polypeptide extract and analyzing the adjustment direction and adjustment amount.

[0059] It should be noted that concentration is to concentrate the purified spleen polypeptide solution to increase the concentration of the spleen polypeptide.

[0060] It should be noted that the concentration in the preparation process of the spleen polypeptide extract in the present invention adopts a reduced pressure concentration method, and the temperature and vacuum degree need to be controlled. The temperature is generally 40°C-60°C, and the vacuum degree is determined according to the performance of the equipment to ensure rapid concentration at a lower temperature without destroying the activity of the spleen polypeptide.

[0061] In another specific embodiment, the spleen polypeptide extract can be concentrated by ultrafiltration concentration or freeze concentration.

[0062] Step 5. Control of drying process parameters for polypeptide extract preparation: Obtain the drying inlet and outlet air temperatures, spray speed and drying time during the preparation of spleen polypeptide extract, determine whether the drying process parameters during the preparation of spleen polypeptide extract meet the requirements, and perform adaptive adjustments.

[0063] As a preferred embodiment, the specific analysis process of step 5 includes: monitoring the drying inlet air temperature, outlet air temperature, spray speed and drying time during the preparation of spleen polypeptide extract, which are respectively recorded as , by analyzing the formula Obtain the compliance coefficient of drying process parameters in the preparation process of spleen polypeptide extract ,in Indicates the correction factor of the preset drying process parameters compliance coefficient, represents a natural constant, They respectively represent the reference air inlet temperature, reference air outlet temperature, reference spray speed and reference drying time during the preparation process of spleen polypeptide extract stored in the database. They respectively represent the influencing factors corresponding to the preset unit inlet temperature deviation, unit outlet temperature deviation, unit spray speed deviation and unit drying time deviation.

[0064] As a preferred embodiment, the specific analysis process of step five also includes: comparing the compliance coefficient of the drying process parameters in the preparation process of the spleen polypeptide extract with the threshold value of the preset drying process parameter compliance coefficient, judging whether the drying process parameters in the preparation process of the spleen polypeptide extract meet the requirements, and if not, judging whether the drying inlet temperature, outlet temperature, spray speed and drying time in the preparation process of the spleen polypeptide extract need to be adjusted and obtaining the adjustment direction and adjustment amount.

[0065] It should be noted that the adaptive adjustment process of the drying process parameters in the preparation process of the spleen polypeptide extract is: comparing the compliance coefficient of the drying process parameters in the preparation process of the spleen polypeptide extract with the preset threshold value of the compliance coefficient of the drying process parameters. If the compliance coefficient of the drying process parameters in the preparation process of the spleen polypeptide extract is less than the preset threshold value of the compliance coefficient of the drying process parameters, the drying process parameters in the preparation process of the spleen polypeptide extract do not meet the requirements.

[0066] According to the drying inlet air temperature, outlet air temperature, spray speed and drying time during the preparation of spleen polypeptide extract and the corresponding reference values, it is determined whether the drying inlet air temperature, outlet air temperature, spray speed and drying time during the preparation of spleen polypeptide extract need to be adjusted and the adjustment direction and adjustment amount are obtained.

[0067] It should be noted that the method for determining whether the drying inlet air temperature, outlet air temperature, spray speed and drying time during the preparation process of spleen polypeptide extract need to be adjusted and obtaining the adjustment direction and adjustment amount is the same as the method for determining whether the temperature of enzymatic hydrolysis during the preparation process of spleen polypeptide extract needs to be adjusted and analyzing the adjustment direction and adjustment amount.

[0068] It should be noted that drying refers to drying the concentrated spleen polypeptide solution to prepare spleen polypeptide powder.

[0069] It should be noted that the drying process of the spleen polypeptide extract in the present invention is spray drying, wherein the key parameters of spray drying include inlet air temperature, outlet air temperature, spray speed and drying time. In a specific embodiment, the inlet air temperature is generally 150°C-200°C, the outlet air temperature is 80°C-100°C, and the drying time is 12-24 hours.

[0070] In another specific embodiment, the spleen polypeptide extract can be dried by freeze drying during preparation.

[0071] In this embodiment, the present invention monitors the process parameters of enzymatic hydrolysis, separation, purification, concentration and drying during the preparation of spleen polypeptide extract, determines whether the requirements are met and performs adaptive adjustments to avoid excessive deviations in the process parameters for preparing spleen polypeptide extract, which may lead to changes in the structure of spleen polypeptide, thereby allowing the spleen polypeptide extract to maintain its natural active conformation and better exert its due efficacy. At the same time, it is beneficial to the optimization and improvement of the preparation process of spleen polypeptide extract and ensure product quality.

[0072] Step 6. Quality inspection and analysis of the finished product of polypeptide extract preparation: The mass spectrometry peptide data of the prepared spleen polypeptide extract and spleen polypeptide injection samples are obtained through mass spectrometry detection technology, and the peak time and source protein type of each high-frequency peptide of the spleen polypeptide extract and spleen polypeptide injection samples are further obtained, and the quality evaluation index of the prepared spleen polypeptide extract is analyzed and feedback is provided.

[0073] As a preferred solution, the specific analysis process of step six includes: F1: detecting the polypeptide sequences contained in the prepared spleen polypeptide extract by mass spectrometry detection technology, summarizing them, and obtaining mass spectrometry peptide data of the prepared spleen polypeptide extract.

[0074] Similarly, the mass spectrometry peptide data of the set spleen polypeptide injection sample is obtained.

[0075] It should be noted that mass spectrometry is a method that can detect protein sequences. It is a method that uses electric and magnetic fields to separate moving ions according to their mass-to-charge ratio and then detects them. By measuring the exact mass of the ions, the compound composition of the ions can be determined.

[0076] F2: According to the mass spectrometry peptide data of the prepared spleen polypeptide extract, the total number of mass spectra matching the peptides of each peptide in the prepared spleen polypeptide extract is obtained, and it is recorded as the PSMs value of each peptide in the prepared spleen polypeptide extract.

[0077] The PSMs value of each peptide segment in the prepared spleen polypeptide extract is compared with the preset PSMs value threshold. If the PSMs value of a peptide segment in the prepared spleen polypeptide extract is greater than the preset PSMs value threshold, the peptide segment is recorded as a high-frequency peptide segment, and the high-frequency peptide segments in the prepared spleen polypeptide extract are counted.

[0078] It should be noted that the PSMs value in the mass spectrum peptide data of spleen polypeptide extract represents the total number of mass spectra matching the peptides, indicating the frequency of peptide detection. In a specific embodiment, peptides with PSMs greater than 10 are recorded as high-frequency peptides.

[0079] Similarly, obtain each high-frequency peptide segment in the spleen polypeptide injection sample.

[0080] F3: Perform high performance liquid chromatography detection on each high-frequency peptide segment in the prepared spleen polypeptide extract to obtain the peak time of each high-frequency peptide segment in the spleen polypeptide extract, and identify the protein type from which each high-frequency peptide segment in the spleen polypeptide extract originates.

[0081] It should be noted that the types of proteins from which the high-frequency peptides of spleen polypeptide extract are derived include but are not limited to: hemoglobin, actin, myosin, cationic trypsin, connexin, ATP synthase, transporter, albumin, cysteine ​​and glycine-rich proteins, methyl mercaptan oxidase, protein S100-A12, protein S100-A9, etc.

[0082] Similarly, the peak time and source protein type of each high-frequency peptide segment of the spleen polypeptide injection sample were obtained.

[0083] As a preferred embodiment, the specific analysis process of step six also includes: constructing protein type sets of high-frequency peptides of spleen polypeptide extract and spleen polypeptide injection sample respectively according to the protein types from which each high-frequency peptide segment of spleen polypeptide extract and spleen polypeptide injection sample is derived, and comparing them with each other to obtain the number of identical protein types of high-frequency peptide segments of spleen polypeptide extract and high-frequency peptide segments of spleen polypeptide injection sample, which is recorded as the number of matching protein types of high-frequency peptide segments of spleen polypeptide extract.

[0084] Substitute the number of matching protein types of high-frequency peptides of spleen polypeptide extract into the preset relationship function between the number of matching protein types of high-frequency peptides of spleen polypeptide extract and the component matching coefficient of spleen polypeptide extract to obtain the component matching coefficient of spleen polypeptide extract.

[0085] Similarly, the peak times of the high-frequency peptides of the spleen polypeptide extract and the spleen polypeptide injection samples are compared with each other to obtain the number of matching peak times of the high-frequency peptides of the spleen polypeptide extract, which is then substituted into the preset relationship function between the number of matching peak times of the high-frequency peptides of the spleen polypeptide extract and the functional compliance coefficient of the spleen polypeptide extract to obtain the functional compliance coefficient of the spleen polypeptide extract.

[0086] The weighted average values ​​of the component conformity coefficient and the functional conformity coefficient of the spleen polypeptide extract are calculated to obtain the quality evaluation index of the prepared spleen polypeptide extract and provide feedback.

[0087] It should be noted that the weights of the component compliance coefficient and the functional compliance coefficient of the spleen polypeptide extract are set values, and the cumulative sum is 1.

[0088] In this embodiment, the present invention detects the polypeptide sequences contained in the spleen polypeptide extract and the spleen polypeptide injection through mass spectrometry analysis technology, identifies the key proteins and polypeptide results of the spleen polypeptide extract and the spleen polypeptide injection, further compares the components and functions of the spleen polypeptide extract and the spleen polypeptide injection, and then evaluates the quality of the prepared spleen polypeptide extract, thereby improving the quality evaluation system of the spleen polypeptide extract and improving the reliability and accuracy of the evaluation results.

[0089] The above contents are merely examples and explanations of the concept of the present invention. Those skilled in the art may make various modifications or additions to the specific embodiments described or replace them in a similar manner. As long as they do not deviate from the concept of the invention or exceed the scope defined by the present invention, they shall all fall within the protection scope of the present invention.

Claims

1. A method for intelligent monitoring and analysis of a polypeptide extract preparation process, characterized in that: The steps include: Step 1: Control of enzymatic hydrolysis process parameters for preparation of polypeptide extracts: obtaining the temperature, duration, pH value and enzyme dosage of the enzymatic hydrolysis during the preparation of the spleen polypeptide extracts, determining whether the enzymatic hydrolysis process parameters during the preparation of the spleen polypeptide extracts meet the requirements, and performing adaptive adjustments; The specific analysis process of step 1 includes: The temperature, duration, pH value and enzyme dosage of the enzymatic hydrolysis process during the preparation of spleen polypeptide extract were monitored and recorded as ; The reference temperature, reference time, reference pH value and reference enzyme dosage of the enzymatic hydrolysis process in the preparation process of the spleen polypeptide extract stored in the extraction database are recorded as ; By analyzing the formula Obtain the compliance coefficient of enzymatic hydrolysis process parameters in the preparation of spleen polypeptide extract ,in Indicates the correction factor of the preset enzymatic hydrolysis process parameters compliance coefficient, They represent the deviation thresholds of the preset enzymatic hydrolysis temperature, duration, pH value and enzyme dosage, respectively. They represent the preset enzymatic hydrolysis temperature, duration, pH value and enzyme dosage weights respectively. ; The specific analysis process of step 1 also includes: D1: Compare the compliance coefficient of the enzymatic hydrolysis process parameters in the preparation process of the spleen polypeptide extract with the preset enzymatic hydrolysis process parameter compliance coefficient threshold. If the compliance coefficient of the enzymatic hydrolysis process parameters in the preparation process of the spleen polypeptide extract is less than the preset enzymatic hydrolysis process parameter compliance coefficient threshold, the enzymatic hydrolysis process parameters in the preparation process of the spleen polypeptide extract do not meet the requirements, and execute D2; D2: comparing the difference between the enzymatic hydrolysis temperature and the reference temperature during the preparation of the spleen polypeptide extract with a preset enzymatic hydrolysis temperature deviation threshold; if the difference between the enzymatic hydrolysis temperature and the reference temperature during the preparation of the spleen polypeptide extract is greater than the preset enzymatic hydrolysis temperature deviation threshold, the enzymatic hydrolysis temperature during the preparation of the spleen polypeptide extract needs to be adjusted, and further obtaining the adjustment direction and adjustment amount of the enzymatic hydrolysis temperature during the preparation of the spleen polypeptide extract; D3: Similarly, according to the analysis method of D2, determine whether the enzymatic hydrolysis time, pH value and enzyme dosage in the preparation process of spleen polypeptide extract need to be adjusted and analyze the adjustment direction and adjustment amount; Step 2: Control of process parameters for separation of polypeptide extract preparation: obtaining the centrifugal speed and centrifugal time for separation during the preparation of spleen polypeptide extract, determining whether the process parameters for separation during the preparation of spleen polypeptide extract meet the requirements, and performing adaptive adjustments; Step 3: Control of process parameters for preparation and purification of polypeptide extracts: obtaining the ionic strength and pH value of the purified eluent during the preparation of the spleen polypeptide extracts, determining whether the process parameters for purification during the preparation of the spleen polypeptide extracts meet the requirements, and performing adaptive adjustments; Step 4: Control of process parameters for preparation and concentration of polypeptide extracts: obtaining the concentration temperature and vacuum degree during the preparation of spleen polypeptide extracts, determining whether the concentration process parameters during the preparation of spleen polypeptide extracts meet the requirements, and performing adaptive adjustments; Step 5, polypeptide extract preparation drying process parameter regulation: obtain the drying inlet and outlet air temperature, spray speed and drying time during the preparation of spleen polypeptide extract, determine whether the drying process parameters during the preparation of spleen polypeptide extract meet the requirements, and perform adaptive adjustment; The specific analysis process of step 5 includes: The inlet air temperature, outlet air temperature, spray speed and drying time of the spleen polypeptide extract preparation process were monitored and recorded as , by analyzing the formula Obtain the compliance coefficient of drying process parameters in the preparation process of spleen polypeptide extract ,in Indicates the correction factor of the preset drying process parameters compliance coefficient, represents a natural constant, They respectively represent the reference air inlet temperature, reference air outlet temperature, reference spray speed and reference drying time during the preparation process of spleen polypeptide extract stored in the database. Respectively represent the influencing factors corresponding to the preset unit inlet air temperature deviation, unit outlet air temperature deviation, unit spray speed deviation and unit drying time deviation; The specific analysis process of step 5 also includes: Compare the compliance coefficient of the drying process parameters in the preparation process of the spleen polypeptide extract with the threshold value of the preset drying process parameter compliance coefficient to determine whether the drying process parameters in the preparation process of the spleen polypeptide extract meet the requirements; if not, determine whether the drying inlet air temperature, outlet air temperature, spray speed and drying time in the preparation process of the spleen polypeptide extract need to be adjusted and obtain the adjustment direction and adjustment amount; Step 6. Quality inspection and analysis of the finished product of polypeptide extract preparation: The mass spectrometry peptide data of the prepared spleen polypeptide extract and spleen polypeptide injection samples are obtained through mass spectrometry detection technology, and the peak time and source protein type of each high-frequency peptide of the spleen polypeptide extract and spleen polypeptide injection samples are further obtained, and the quality evaluation index of the prepared spleen polypeptide extract is analyzed and feedback is provided.

2. The method for intelligent monitoring and analysis of a polypeptide extract preparation process according to claim 1, characterized in that: The specific analysis process of step 2 is as follows: Monitor the centrifugal speed and centrifugal time of separation during the preparation of the spleen polypeptide extract, and extract the reference centrifugal speed and reference centrifugal time of separation during the preparation of the spleen polypeptide extract stored in the database, compare them, and obtain the difference between the centrifugal speed and centrifugal time of separation during the preparation of the spleen polypeptide extract, substitute it into the relationship function between the difference between the centrifugal speed and centrifugal time of separation and the compliance coefficient of the separation process parameters, and obtain the compliance coefficient of the separation process parameters during the preparation of the spleen polypeptide extract; According to the compliance coefficient of the separation process parameters in the preparation process of the spleen polypeptide extract, it is judged whether the separation process parameters in the preparation process of the spleen polypeptide extract meet the requirements. If they do not meet the requirements, it is judged whether the centrifugal speed and centrifugal time of the separation in the preparation process of the spleen polypeptide extract need to be adjusted and the adjustment direction and adjustment amount are obtained.

3. The method for intelligent monitoring and analysis of a polypeptide extract preparation process according to claim 1, characterized in that: The specific analysis process of step three is as follows: Monitor the ionic strength and pH value of the purified eluent during the preparation of the spleen polypeptide extract, and compare them with the ionic strength and pH value of the reference eluent purified during the preparation of the spleen polypeptide extract stored in the database to analyze the compliance coefficient of the purification process parameters during the preparation of the spleen polypeptide extract; According to the compliance coefficient of the purification process parameters in the preparation process of the spleen polypeptide extract, it is judged whether the purification process parameters in the preparation process of the spleen polypeptide extract meet the requirements. If they do not meet the requirements, it is judged whether the ionic strength and pH value of the purified eluent in the preparation process of the spleen polypeptide extract need to be adjusted and the adjustment direction and adjustment amount are obtained.

4. The method for intelligent monitoring and analysis of a polypeptide extract preparation process according to claim 1, characterized in that: The specific analysis process of step 4 is as follows: Monitor the concentration temperature and vacuum degree during the preparation of spleen polypeptide extracts, and compare them with the reference temperature and reference vacuum degree during the preparation of spleen polypeptide extracts stored in the database, and analyze the compliance coefficient of the concentration process parameters during the preparation of spleen polypeptide extracts; According to the compliance coefficient of the concentration process parameters in the preparation process of the spleen polypeptide extract, it is judged whether the concentration process parameters in the preparation process of the spleen polypeptide extract meet the requirements. If they do not meet the requirements, it is judged whether the concentration temperature and vacuum degree in the preparation process of the spleen polypeptide extract need to be adjusted and the adjustment direction and adjustment amount are obtained.

5. The method for intelligent monitoring and analysis of a polypeptide extract preparation process according to claim 1, characterized in that: The specific analysis process of step six includes: F1: Detect the polypeptide sequences contained in the prepared spleen polypeptide extract by mass spectrometry detection technology, summarize them, and obtain the mass spectrometry peptide data of the prepared spleen polypeptide extract; Similarly, the mass spectrometry peptide data of the set spleen polypeptide injection sample is obtained; F2: According to the mass spectrometry peptide data of the prepared spleen polypeptide extract, the total number of mass spectra matching the peptides of each peptide in the prepared spleen polypeptide extract is obtained, and it is recorded as the PSMs value of each peptide in the prepared spleen polypeptide extract; The PSMs value of each peptide segment in the prepared spleen polypeptide extract is compared with a preset PSMs value threshold value. If the PSMs value of a certain peptide segment in the prepared spleen polypeptide extract is greater than the preset PSMs value threshold value, the peptide segment is recorded as a high-frequency peptide segment, and each high-frequency peptide segment in the prepared spleen polypeptide extract is counted; Similarly, obtain each high-frequency peptide segment in the spleen polypeptide injection sample; F3: Perform high performance liquid chromatography detection on each high-frequency peptide segment in the prepared spleen polypeptide extract to obtain the peak time of each high-frequency peptide segment in the spleen polypeptide extract, and identify the protein type from which each high-frequency peptide segment in the spleen polypeptide extract originates; Similarly, the peak time and source protein type of each high-frequency peptide segment of the spleen polypeptide injection sample were obtained.

6. The method for intelligent monitoring and analysis of a polypeptide extract preparation process according to claim 5, characterized in that: The specific analysis process of step six also includes: According to the protein types of the high-frequency peptides from the spleen polypeptide extract and the spleen polypeptide injection samples, protein type sets of the high-frequency peptides from the spleen polypeptide extract and the spleen polypeptide injection samples were constructed respectively, and compared with each other to obtain the number of identical protein types of the high-frequency peptides from the spleen polypeptide extract and the high-frequency peptides from the spleen polypeptide injection samples, which was recorded as the number of identical protein types of the high-frequency peptides from the spleen polypeptide extract; Substituting the number of matching protein types of the high-frequency peptide segments of the spleen polypeptide extract into a preset relationship function between the number of matching protein types of the high-frequency peptide segments of the spleen polypeptide extract and the component matching coefficient of the spleen polypeptide extract, to obtain the component matching coefficient of the spleen polypeptide extract; Similarly, the peak times of the high-frequency peptides of the spleen polypeptide extract and the spleen polypeptide injection sample are compared with each other to obtain the number of coincident peak times of the high-frequency peptides of the spleen polypeptide extract, which is substituted into the preset relationship function between the number of coincident peak times of the high-frequency peptides of the spleen polypeptide extract and the functional conformity coefficient of the spleen polypeptide extract to obtain the functional conformity coefficient of the spleen polypeptide extract; The weighted average values ​​of the component conformity coefficient and the functional conformity coefficient of the spleen polypeptide extract are calculated to obtain the quality evaluation index of the prepared spleen polypeptide extract and provide feedback.

Images