Method for detecting ethylene bis oleamide leached substance in medical instrument

Abstract

The invention discloses a method for detecting an ethylene bis oleamide leached substance in a medical instrument. The method is not only suitable for production quality control of the medical apparatus, but also can be used for market supervision and management of the medical apparatus, and an effective technical means is provided for guaranteeing the safety of the medical apparatus. The invention provides a detection method based on a high performance liquid chromatograph (HPLC), which is not only suitable for production quality control of the medical apparatus, but also can be used for market supervision and management of the medical apparatus, and provides an effective technical means for guaranteeing the safety of the medical apparatus. The invention provides a detection method based on a high performance liquid chromatograph (HPLC), and through specific sample pretreatment and optimized chromatographic conditions, high-sensitivity and high-accuracy detection of the ethylene bis oleamide leached substance in the medical instrument is realized. And detailed methodological verification is carried out. The method has the characteristics of low detection limit, simplicity and convenience in pretreatment, high analysis speed, high repeatability, good accuracy and the like.

Description

Technical Field

[0001] The invention belongs to the technical field of ethylenebisoleamide leachable matter detection, and specifically relates to a method for detecting ethylenebisoleamide leachable matter in medical devices. Background Art

[0002] With the rapid development of the medical device industry, the concern about the safety of medical devices is also increasing. Ethylene bisoleamide (EBO) is a good lubricant with good opening performance and relatively small dosage. It can reduce the melt index and improve the appearance quality of the product. It is easy to disperse and has low migration. It is used as a lubricant, lubricant, anti-adhesive agent and release agent in plastics, and an antistatic agent for cellophane. It is suitable for internal and external lubricants of ABS, polystyrene, polyvinyl chloride, nylon, cellulose acetate, polyvinyl acetate and phenolic resin. It is widely used in a variety of medical device materials to improve their physical and chemical properties. However, the residues of ethylene bisoleamide may pose a potential risk to human health, especially when these devices are used for long-term implantation or direct contact with blood and other body fluids. Therefore, accurate and effective detection of EBO residues in medical devices has become an important part of ensuring the safety of medical devices.

[0003] At present, there is a lack of methods for detecting EBO residues in medical devices, and there is little relevant testing technical information.

[0004] Therefore, there is an urgent need for a new detection method that can provide a faster, simpler, more sensitive and specific EBO residue detection to meet the actual needs of medical device production and supervision. Summary of the invention

[0005] The object of the present invention is to provide a method for detecting leachable matter of ethylenebisoleamide in medical devices in order to solve the above-mentioned problems.

[0006] The technical solution adopted by the present invention is as follows: a method for detecting leachable substances of ethylenebisoleamide in medical devices, the detection method comprising the following steps:

[0007] S1: Prepare the instrument: high performance liquid chromatography (HPLC);

[0008] S2: Prepare the test solution: Take one sample, add 50.0 mL of methanol at a ratio of 0.2 g / mL, extract at 37°C ± 1°C for 72 ± 2 h, separate the sample from the liquid, cool the solution to room temperature, and prepare 2 portions of each extraction solvent in parallel as the sample test solution;

[0009] S3: Prepare blank sample solution: Take the same volume of methanol from the same batch, except without adding sample and refer to the test solution treatment method to obtain blank sample solution;

[0010] S4: Preparation of EBO standard stock solution A: Accurately weigh 10.55 mg (95.0%) of EBO, add dichloromethane to prepare an EBO standard stock solution with a concentration of 1002.25 μg / mL;

[0011] S5: Preparation of EBO standard stock solution B: Accurately pipette 2.5 mL of EBO standard stock solution A into a 25 mL volumetric flask, add methanol to dilute to the mark, and shake well to obtain 100 μg / mL standard stock solution B;

[0012] S6: Prepare linear solutions: accurately pipette appropriate amount of EBO standard stock solution B, dilute with methanol, and prepare 5 concentration gradient standard curve solutions, which are recorded as STD1, STD2, STD3, STD4 and STD5;

[0013] S7: Prepare detection limit solution: prepare 0.1 μg / mL EBO standard solution. The signal-to-noise ratio of the response value of the solution shall not be less than 3:1.

[0014] S8: Prepare the quantitative limit solution: prepare a 0.2 μg / mL EBO solution. The signal-to-noise ratio of the response value of the solution must not be less than 10:1. The concentration corresponding to the solution is used as the quantitative limit.

[0015] S9: Accuracy Solution:

[0016] By using the sample spike method, add an appropriate amount of standard solution to the test solution to prepare three accuracy solutions at low, medium and high concentration levels, and prepare three replicates at each concentration;

[0017] S10: Repeatability solution: Take 9 portions of accuracy solution to obtain;

[0018] S11: HPLC analysis is used to determine the content of ethylenebisoleamide leachable matter in the sample, and the data results obtained from the analysis are recorded and saved, and then the ethylenebisoleamide detection process can be ended.

[0019] In a preferred embodiment, in step S1, the mobile phase is acetonitrile and water, and the ratio of the mobile phase is 7:3.

[0020] In a preferred embodiment, in step S1, the instrument is prepared by adjusting various parameters to a working state: the flow rate is 1 mL / min; the chromatographic column is a XDB-C18 5.0 μm 4.6 mm×250 mm chromatographic column.

[0021] In a preferred embodiment, in step S1, the column temperature is 30°C, the injection volume is 10 μL, and the detection wavelength is 204 nm.

[0022] In a preferred embodiment, in step S2, 50.0 mL of methanol is added at a ratio of 0.2 g / mL.

[0023] In a preferred embodiment, in steps S7 and S8, the concentration corresponding to the detection limit solution is used as the detection limit, and the concentration corresponding to the quantification limit solution is used as the quantification limit.

[0024] In a preferred embodiment, in step S9, a sample spike method is adopted to add an appropriate amount of standard solution to the test solution to prepare accuracy solutions at three concentration levels of low, medium and high, and three replicates are prepared in parallel for each concentration.

[0025] In a preferred embodiment, in step S10, the repeatability solution is obtained by taking 9 portions of the accuracy solution.

[0026] In a preferred embodiment, in step S11, ethylenebisoleamide has a maximum absorption peak at 204 nm, with a peak at 3.3 min, and a good peak shape.

[0027] In summary, due to the adoption of the above technical solution, the beneficial effects of the present invention are:

[0028] The present invention is not only applicable to the production quality control of medical devices, but also can be used for the market supervision and management of medical devices, providing an effective technical means to ensure the safety of medical devices. The invention provides a detection method based on high performance liquid chromatography (HPLC), which achieves high sensitivity and high accuracy detection of ethylenebisoleamide leachable substances in medical devices through specific sample pretreatment and optimized chromatographic conditions. A detailed methodological verification was carried out. The method has a low detection limit, simple pretreatment, fast analysis speed, high repeatability, good accuracy and the like. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 It is the standard spectrogram of the present invention;

[0030] Figure 2 This is the methanol blank solution spectrum of the present invention;

[0031] Figure 3 It is the spectrum of the test solution in the present invention;

[0032] Figure 4 It is the curve spectrum in the present invention. DETAILED DESCRIPTION

[0033] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0034] Reference Figure 1-4 ,

[0035] A method for detecting leachable matter of ethylenebisoleamide in medical devices, the method comprising the following steps:

[0036] The following steps are involved:

[0037] 1) Instrument preparation: High performance liquid chromatography (HPLC)

[0038] 2) Test solution: Take one sample from each group and add 50.0 mL of extraction solvent at a ratio of 0.2 g / mL. Extract at 37°C±1°C for 72±2 h. Take the extraction solution for later use. Prepare 2 portions in parallel as sample test solutions.

[0039] 3) Blank sample solution: Take the same volume of methanol from the same batch, without adding sample and refer to the test solution treatment method to obtain a blank sample solution.

[0040] 4) Ethylene bisoleamide standard stock solution A: Accurately weigh 10.55 mg (95.0%) of ethylene bisoleamide and add toluene to prepare an ethylene bisoleamide standard stock solution with a concentration of 1002.25 μg / mL.

[0041] 5) Ethylene bisoleamide standard stock solution B: Accurately pipette 2.5 mL of EBO standard stock solution A into a 25 mL volumetric flask, add methanol to dilute to the scale, and shake well to obtain 100 μg / mL standard stock solution B.

[0042] 6) Linear solution: Accurately pipette appropriate amount of EBO standard stock solution B, dilute with methanol, and prepare 5 concentration gradient standard curve solutions, recorded as STD1, STD2, STD3, STD4 and STD5.

[0043] 7) Detection limit solution: Prepare a 0.1 μg / mL EBO standard solution. The signal-to-noise ratio of the response value of the solution must not be less than 3:1. The concentration corresponding to the solution is used as the detection limit.

[0044] 8) Quantitation limit solution: Prepare a 0.2 μg / mL EBO solution. The response value signal-to-noise ratio of this solution must not be less than 10:1. The concentration corresponding to this solution is used as the quantitation limit.

[0045] 9) Repeatability solution: Use the sample spike method to add an appropriate amount of standard solution to the test solution to prepare accuracy solutions at three concentration levels: low, medium and high. Prepare three replicates for each concentration.

[0046] 10) Accuracy solution: Prepare three solutions with different concentration levels: low, medium, and high as accuracy solutions.

[0047] (1) Accurately pipette 35 μL of ethylenebisoleamide standard stock solution B and add 9.965 mL of methanol. Extract at 37°C ± 1°C for 72 h and then cool to obtain accuracy solution 1. Prepare three replicates in parallel.

[0048] (2) Accurately pipette 0.1 mL of ethylenebisoleamide standard stock solution B and add 9.9 mL of methanol. Extract at 37°C ± 1°C for 72 h and then cool to obtain accuracy solution 2. Prepare three replicates in parallel.

[0049] (3) Accurately pipette 0.23 mL of ethylenebisoleamide standard stock solution B and add 9.77 mL of methanol. Extract at 37°C ± 1°C for 72 h and then cool to obtain accuracy solution 3. Prepare three replicates in parallel.

[0050] 11) HPLC analysis was used to determine the content of ethylenebisoleamide in the sample.

[0051] In step 1), the instrument is prepared by adjusting various parameters to the working state: the flow rate is 1 mL / min; the chromatographic column is XDB-C18 5.0 μm 4.6 mm×250 mm chromatographic column;

[0052] In step 1), the column temperature was 30°C, the injection volume was 10 μL, and the detection wavelength was 204 nm.

[0053] The disposable central venous catheter and the kit can leach a substance into methanol.

[0054] The present invention belongs to the detection of known leachable substances in medical devices. The present invention adopts a gas chromatography-mass spectrometry (HPLC) to determine a method for determining ethylenebisoleamide in medical device leachable substances, and performs detailed methodological verification. The method has a low detection limit, simple pretreatment, fast analysis speed, high repeatability, good accuracy and the like. The method provides a feasible method for detecting ethylenebisoleamide in medical device leachable substances.

[0055] Disposable central venous catheter and kit embodiment 1:

[0056] Instruments and reagents

[0057] High performance liquid chromatography (Agilent Technologies (China) Co., Ltd. 1260Ⅱ with DAD detector), electronic balance (accuracy 0.0001g, Mettler Toledo, Switzerland), ethylene bisoleamide (EBO) (100 mg / bottle, Shanghai Yuanye Biotechnology Co., Ltd.), methanol (4 L / bottle, Pormee)

[0058] Instrument conditions

[0059] The ethylene bisoleamide HPLC parameter settings are shown in Table 1.

[0060] Table 1 HPLC instrument conditions

[0061] Parameter name Specific parameters Chromatographic columns XDB-C185μm4.6mm×250mm Column temperature 30℃ Mobile phase Acetonitrile: Water (7:3) Injection volume 10μL Detection wavelength 204nm Flow rate 1.0mL / min

[0062] 1. Solution preparation

[0063] 1) Test solution: Take one sample from each sample and add 50 mL of methanol at a ratio of 0.2 g / mL. Extract for 72 h at 37°C ± 1°C to simulate the clinical use temperature. Separate the sample from the liquid and cool the solution to room temperature. Prepare 2 portions of each extraction solvent in parallel as the sample test solution.

[0064] 2) Blank sample solution: Take the same volume of methanol from the same batch, without adding sample and refer to the test solution treatment method to obtain a blank sample solution.

[0065] 3) Ethylene bisoleamide standard stock solution A: Accurately weigh 10.55 mg (95%) of ethylene bisoleamide and add methanol to prepare an ethylene bisoleamide standard stock solution with a concentration of 1002.25 μg / mL.

[0066] 4) Ethylene bisoleamide standard stock solution B: Accurately pipette 2.5 mL of ethylene bisoleamide standard stock solution A into a 10 mL volumetric flask, add methanol to dilute to the mark, and shake well to obtain a 100 μg / mL ethylene bisoleamide standard stock solution.

[0067] 5) Linear solution: Accurately pipette appropriate amount of ethylenebisoleamide standard stock solution, dilute with methanol, and prepare 5 concentration gradient standard curve solutions, recorded as STD1, STD2, STD3, STD4 and STD5.

[0068] 6) Detection limit solution: Prepare a 0.1 μg / mL ethylenebisoleamide standard solution. The signal-to-noise ratio of the response value of the solution should not be less than 3:1. The concentration corresponding to the solution is used as the detection limit.

[0069] 7) Quantitation limit solution: Prepare a 0.2 μg / mL ethylenebisoleamide solution. The signal-to-noise ratio of the response value of the solution should not be less than 10:1. The concentration corresponding to the solution is used as the quantitation limit.

[0070] 8) Repeatability solution: Use the sample spike method to add an appropriate amount of standard solution to the test solution to prepare accuracy solutions at three concentration levels: low, medium and high. Prepare three replicates for each concentration.

[0071] 9) Accuracy solution: Prepare three solutions with different concentration levels: low, medium, and high as accuracy solutions.

[0072] (1) Accurately pipette 35 μL of ethylenebisoleamide standard stock solution B and add 9.965 mL of methanol. Extract at 37°C ± 1°C for 72 h and then cool to obtain accuracy solution 1. Prepare three replicates in parallel.

[0073] (2) Accurately pipette 0.1 mL of ethylenebisoleamide standard stock solution B and add 9.9 mL of methanol. Extract at 37°C ± 1°C for 72 h and then cool to obtain accuracy solution 2. Prepare three replicates in parallel.

[0074] (3) Accurately pipette 0.23 mL of ethylenebisoleamide standard stock solution B and add 9.77 mL of methanol. Extract at 37°C ± 1°C for 72 h and then cool to obtain accuracy solution 3. Prepare three replicates in parallel.

[0075] 2. Method Validation

[0076] 4.1. Exclusivity

[0077] The reagent blank solution, blank sample solution, test solution and ethylenebisoleamide standard solution (1.0 μg / mL) were tested on the machine respectively, and the chromatograms were recorded. Figure 1 to Figure 4 It can be seen that there is no interfering peak near the target peak (ethylene bisoleamide) in the ethylene bisoleamide standard solution, blank solution, and test solution. Specific test results are shown in Table 2. From the above results, it can be seen that this method has good specificity.

[0078] Table 2 Specificity results of ethylene bisoleamide test

[0079] name EBO standard solution blank solution Test solution Interference peak none none none Peak area 52.660 Not detected Not detected

[0080] 4.2. Linear

[0081] The blank solution and the linear solution were tested. The results showed that the correlation coefficient of the linear equation of the target peak was 0.9995. The specific results are shown in Table 3. From the above results, it can be seen that this method has good linearity.

[0082] Table 3 EBO linear test results

[0083]

[0084] 4.3 Limit of detection and limit of quantification

[0085] The detection limit and quantification limit solutions of ethylenebisoleamide were tested on the machine according to the conditions in Table 1. The solution corresponding to the signal-to-noise ratio of the response value of ethylenebisoleamide must not be lower than 3:1 is the detection limit; the solution corresponding to the signal-to-noise ratio of the response value must not be lower than 10:1 is the quantification limit; the specific results are shown in Table 4.

[0086] Table 4 Results of detection limit and quantification limit of EBO

[0087]

[0088] 4.4. Repeatability and accuracy

[0089] Take nine accuracy solutions in three groups of low, medium and high, test them according to the conditions in Table 1, and record the chromatograms. According to the fourth guideline 9101 of the 2020 edition of the Chinese Pharmacopoeia, the acceptable range of target recovery in the solution is 80% to 115%. Repeatability is expressed as the RSD value of the recovery rate of 9 solutions, and the RSD should be ≤6%. The test results are shown in Table 5. It can be seen from the results in Table 5 that this method has good accuracy and repeatability.

[0090] Table 5 EBO repeatability test results

[0091]

[0092] 5. EBO content test results

[0093] The test was performed using HPLC, and the EBO test results are shown in Table 7.

[0094] Table 7 EBO content test results

[0095]

[0096] The present invention is not only applicable to the production quality control of medical devices, but also can be used for the market supervision and management of medical devices, providing an effective technical means to ensure the safety of medical devices. The invention provides a detection method based on high performance liquid chromatography (HPLC), which achieves high sensitivity and high accuracy detection of ethylenebisoleamide leachable substances in medical devices through specific sample pretreatment and optimized chromatographic conditions. A detailed methodological verification was carried out. The method has a low detection limit, simple pretreatment, fast analysis speed, high repeatability, good accuracy and the like.

[0097] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, the elements defined by the sentence "comprise a ..." do not exclude the existence of other identical elements in the process, method, article or device including the elements.

[0098] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, a person skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for detecting leachable matter of ethylenebisoleamide in medical devices, characterized in that: The detection method comprises the following steps: S1: Prepare the instrument: high performance liquid chromatography (HPLC); S2: Prepare the test solution: Take one sample, add 50.0 mL of methanol at a ratio of 0.2 g / mL, extract at 37°C ± 1°C for 72 ± 2 h, separate the sample from the liquid, cool the solution to room temperature, and prepare 2 portions of each extraction solvent in parallel as the sample test solution; S3: Prepare blank sample solution: Take the same volume of methanol from the same batch, except without adding sample and refer to the test solution treatment method to obtain blank sample solution; S4: Preparation of EBO standard stock solution A: Accurately weigh 10.55 mg (95.0%) of EBO, add dichloromethane to prepare an EBO standard stock solution with a concentration of 1002.25 μg / mL; S5: Preparation of EBO standard stock solution B: Accurately pipette 2.5 mL of EBO standard stock solution A into a 25 mL volumetric flask, add methanol to dilute to the mark, and shake well to obtain 100 μg / mL standard stock solution B; S6: Prepare linear solutions: accurately pipette appropriate amount of EBO standard stock solution B, dilute with methanol, and prepare 5 concentration gradient standard curve solutions, which are recorded as STD1, STD2, STD3, STD4 and STD5; S7: Prepare detection limit solution: prepare 0.1 μg / mL EBO standard solution. The signal-to-noise ratio of the response value of the solution shall not be less than 3:

1. S8: Prepare the quantitative limit solution: prepare a 0.2 μg / mL EBO solution. The signal-to-noise ratio of the response value of the solution must not be less than 10:

1. The concentration corresponding to the solution is used as the quantitative limit. S9: Accuracy Solution: By using the sample spike method, add an appropriate amount of standard solution to the test solution to prepare three accuracy solutions at low, medium and high concentration levels, and prepare three replicates at each concentration; S10: Repeatability solution: Take 9 portions of accuracy solution to obtain; S11: HPLC analysis is used to determine the content of ethylenebisoleamide leachable matter in the sample, and the data results obtained from the analysis are recorded and saved, and then the ethylenebisoleamide detection process can be ended.

2. A method for detecting leachable matter of ethylenebisoleamide in a medical device according to claim 1, characterized in that: In step S1, the mobile phases used were acetonitrile and water in a ratio of 7:3, and the column temperature was 30°C.

3. A method for detecting leachable matter of ethylenebisoleamide in a medical device according to claim 1, characterized in that: In step S1, the instrument is prepared by adjusting various parameters to a working state, the flow rate is 1 mL / min, and the chromatographic column is XDB-C18 5.0 μm 4.6 mm×250 mm.

4. A method for detecting leachable matter of ethylenebisoleamide in a medical device according to claim 1, characterized in that: In the step S1, the injection volume is 10 μL and the detection wavelength is 204 nm.

5. The method for detecting leachable matter of ethylenebisoleamide in a medical device according to claim 1, characterized in that: In the step S2, 50.0 mL of methanol is added at a ratio of 0.2 g / mL.

6. A method for detecting leachable matter of ethylenebisoleamide in a medical device according to claim 1, characterized in that: In the steps S7 and S8, the concentration corresponding to the detection limit solution is used as the detection limit, and the concentration corresponding to the quantification limit solution is used as the quantification limit.

7. A method for detecting leachable matter of ethylenebisoleamide in a medical device according to claim 1, characterized in that: In step S9, a sample spike method is adopted to add an appropriate amount of standard solution to the test solution to prepare three accuracy solutions at low, medium and high concentration levels, and three replicates are prepared in parallel for each concentration.

8. The method for detecting leachable matter of ethylenebisoleamide in a medical device according to claim 1, characterized in that: In step S10, 9 portions of the accuracy solution are taken to obtain the repeatability solution.

Images