Method for detecting biological product diluent by using ion chromatography

Abstract

The invention discloses a method for detecting a biological product diluent by using ion chromatography, and belongs to the field of analysis and detection. The ion chromatography detection technology is adopted to analyze and detect the biological product diluents, the result shows that the diluents with different components can be effectively separated on an ion chromatogram, qualitative and quantitative analysis can be performed on the diluents with different components by performing chromatographic analysis and detection on the diluents, and through comparative analysis, the content of the diluents with different components can be determined. The smaller the relative standard deviation of the retention time and area of the diluent is, the better the uniformity of the diluent is. According to the method, the ion condition in the diluent can be well evaluated, powerful technical support can be provided for the product by guiding and screening the diluent and optimizing the production process, and meanwhile, the method has profound significance on further comprehensive evaluation of the product quality by combining ion components in the diluents of various manufacturers in the industry with other indexes.

Description

Technical Field

[0001] The invention relates to a method for detecting a biological product diluent by utilizing ion chromatography, and belongs to the field of analysis and detection. Background Art

[0002] Dilution, as the name implies, is a mixed solution obtained by adding water (or other solvents) to the pure solution we need to reduce the concentration. In daily practical experimental operations, we often do not need pure solutions for experiments, so we use water dilution to obtain solutions with lower concentrations, so as to conduct better experiments. In summary: any solution obtained by adding solvents to reduce the concentration is a dilution. At present, in the biological products industry, due to different manufacturers and functions, biological product dilutions are diversified. The dilution components of different manufacturers are different, and the components of different dilutions of the same manufacturer are also different. Different dilutions are usually only effective for specific samples, and may not be effective for samples from other sources. The use of unstable diluents cannot play a good dilution role, and may also produce adverse reactions, affecting the decline in product efficacy, weakened immune protection, increased side effects after vaccination, increased instability, and difficulty in controlling product batch differences, which directly have an important impact on the stability and effectiveness of the product. Therefore, relevant personnel should pay full attention to the selection and use of diluents, and make reasonable choices, so strict control is required.

[0003] The detection methods of diluent mainly include the following: Chromatographic analysis, Karl Fischer method, density meter method, headspace-gas chromatography, acid-base titration method, diluent is a commonly used sample processing reagent, but its selection and use method also have an important impact on sample stability. Therefore, when selecting and using diluent, it is necessary to ensure the uniformity and stability of the results. Summary of the invention

[0004] The purpose of the present invention is to provide a method for detecting diluents of biological products using ion chromatography, in view of the shortcomings and technical gaps in the analysis and detection of diluents in biological products by ion chromatography. The method can effectively analyze the distribution and content of components in the diluent, and when selecting and using the diluent, the uniformity and stability of the results can be ensured.

[0005] A method for detecting a biological product dilution using ion chromatography comprises the following steps: (1) Qualitative analysis: The biological product dilution was analyzed and detected by ion chromatography to obtain the ion chromatogram of the dilution in the biological product. The ion chromatography system used was DIONEX ICS-1600; suppressor was DIONEX ARS-300; automatic sampler was DIONEX AS40; the ion chromatography analysis conditions were as follows: Chromatographic columns: IonPac AG23 Guard 4×50mm, IonPac AS23 Analytical 4×250mm; eluent: 0.8mMNaHCO3 solution and 4.5mMNa2CO3 solution; flow rate: 1.0mL / min; injection volume: 25μL; pressure limit: 200-3000psi; sampling time: 30min; cell temperature: 35℃; column temperature: 33℃; current value: 25mA; qualitative analysis is performed based on the retention time of the chromatographic peaks of the ion chromatogram to determine the components contained in the diluent of the biological product and to determine the uniformity of the diluent.

[0006] (2) Quantitative analysis: The chromatographic peaks in the ion chromatogram are analyzed, and the chromatographic peak areas corresponding to the diluent components with different retention times are processed to obtain the area results corresponding to the retention time of each peak of the diluent component. Through comparative analysis, the effective content of a certain substance in different diluents can be evaluated, guidance can be provided for the screening of diluent uniformity, key information can be provided for the product production process, and the production process can be optimized. After repeated verification, this method is simple to operate, accurate in results, highly sensitive, and has achieved satisfactory results in actual detection.

[0007] The results show that this method can better evaluate the ion situation in the diluent. Through the effective separation of the diluent on the ion chromatogram, it can be concluded that the components of different diluents are different in retention time and area. Through comparative analysis and control of ion chromatography detection technology, the relative standard deviation of sample retention time and area is reduced, further ensuring the uniformity and stability of the diluent in the application of biological products. This method can be used to guide the screening of raw materials, optimize the production process, and provide key information for products. At the same time, the ion composition in the diluent of each biological product manufacturer in the industry can be benchmarked, combined with other indicators, and the quality of the diluent can be further comprehensively evaluated, providing technical support for product quality control, which is of far-reaching significance.

[0008] Compared with the prior art, the present invention has the following beneficial effects: The establishment of this method provides a technology for detecting components in biological product diluents using ion chromatography. Its detection method is a very important indicator in diluent testing, which enables the ion components of different diluents to be effectively separated on the ion chromatogram. After repeated verification, this method is simple to operate, accurate in results, highly sensitive, and has achieved satisfactory results in actual testing. By comparison, diluents with small batch differences, high stability and uniformity were screened out to guide production, while improving the uniformity and stability of the product. DETAILED DESCRIPTION

[0009] The following is an explanation of the establishment of an analytical method for detecting dilutions in biological products using the ion chromatography method of the present invention through a specific implementation. Example

[0010] The biological product dilution was analyzed and detected by an ion chromatograph to obtain an ion chromatogram of the dilution in the biological product. The ion chromatographic system used was: DIONEX ICS-1600; suppressor: DIONEX ARS-300; automatic sampler: DIONEXAS40; and the ion chromatographic analysis conditions were as follows: Chromatographic columns: IonPac AG23 Guard 4×50mm, IonPac AS23 Analytical 4×250mm; eluent: NaHCO3 solution and Na2CO3 solution; flow rate: 1.0mL / min; injection volume: 25μL; pressure limit: 200-3000psi; sampling time: 30min; cell temperature: 35℃; column temperature: 33℃; current value: 25mA. The concentration of NaHCO3 solution is 0.8mM; the concentration of Na2CO3 solution is 4.5mM.

[0011] The ICS-1600 ion chromatograph uses isocratic elution. The sample is pushed into the analytical column by the eluent. The ion exchange separation method is adopted. According to the difference in ion radius and valence state, the sample passes through the separation column and enters the suppressor. The conductivity of the eluent is suppressed and the background noise is reduced. The conductivity cell detects the conductivity of the sample and transmits the detection signal to the data collection system to obtain the ion chromatogram of the biological product dilution.

[0012] After the diluent is separated by chromatography, a series of chromatographic peaks can be obtained. Each chromatographic peak represents a substance. After measuring the diluent of a manufacturer, the content of each component in the diluent can be accurately determined based on the retention time and area. After repeated tests, the diluent samples of the same manufacturer have good repeatability. The components contained are chloride, chlorate, phosphate, and the relative standard deviation of retention time and area is uniform and stable, and the results are basically consistent (see Table 1); the components of the diluent samples of different manufacturers vary greatly, and the uniformity and stability are not good (see Table 2).

[0013] In actual application, it is difficult to infer the component results of a certain diluent. After ion chromatography analysis and detection by the method of the present invention, it can be combined with reality, and the selection and dilution process of the diluent can be well controlled in the experiment and production process, and a diluent with relatively stable components can be screened out, which is very important for the uniformity and stability of products of biological product companies during the production process.

Claims

1. A method for detecting a biological product dilution using ion chromatography, comprising the following steps: (1) Qualitative analysis: The biological product dilution was analyzed and detected by ion chromatography to obtain the ion chromatogram of the dilution in the biological product. The ion chromatography system used was DIONEX ICS-1600; suppressor was DIONEX ARS-300; automatic sampler was DIONEX AS40; the ion chromatography analysis conditions were as follows: Chromatographic columns: IonPac AG23 Guard 4×50mm, IonPac AS23 Analytical 4×250mm; eluent: NaHCO3 solution and Na2CO3 solution; flow rate: 1.0mL / min; injection volume: 25μL; pressure limit: 200-3000psi; sampling time: 30min; cell temperature: 35℃; column temperature: 33℃; current value: 25mA; Conduct qualitative analysis based on the retention time of the chromatographic peaks in the ion chromatogram to determine the components contained in the dilution of the biological product; (2) Quantitative analysis: Analyze the chromatographic peaks in the ion chromatogram, process the chromatographic peak areas corresponding to the dilution components at different retention times, obtain the peak areas corresponding to the dilution components, and determine the content of the components in the dilution.

2. The method for detecting diluent in biological products by ion chromatography according to claim 1, characterized in that: The concentration of NaHCO3 solution is 0.8 mM, and the concentration of Na2CO3 solution is 4.5 mM.

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