CN114062535B - Method for detecting content of volatile and semi-volatile unknown substances in medical instrument leaching liquor - Google Patents
Method for detecting content of volatile and semi-volatile unknown substances in medical instrument leaching liquor Download PDFInfo
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Abstract
The invention discloses a method for detecting the content of volatile and semi-volatile unknown substances in medical instrument leaching liquor, which comprises the following steps: firstly, preparing a series of mixed solutions of standard working solution and a substitute standard substance, analyzing by using a gas chromatography-mass spectrometer, establishing a standard working solution calibration curve according to an analysis result, and calculating a relative response factor of the substitute standard substance relative to dimethyl phthalate; secondly, leaching the medical instrument to obtain a leaching solution, detecting the leaching solution by using a gas chromatography-mass spectrometer, respectively comparing and identifying the detection result of the leaching solution with the detection results of the series of standard working solutions and the mixed solution of the substitute standard substance, and calculating to obtain the concentration of an unknown substance in the leaching solution; and finally, calculating according to the concentration of the unknown substance to obtain the content of the unknown substance. The detection method provided by the invention reduces the safety risk brought by only paying attention to the quantification of the volatile and semi-volatile known substances, and also reduces the cost for developing and verifying various volatile and semi-volatile known substances.
Description
Technical Field
The invention relates to the technical field of analysis method detection, in particular to a method for detecting the content of volatile and semi-volatile unknown substances in medical apparatus leaching liquor.
Background
During the process of the medical device product continuously contacting and functioning with the human body, or when interacting with other media (such as liquid medicine, blood, etc.) in use, some chemicals are released, which are collectively called leachable substances, which generally include sterilization residues, processing residues, degradation products, solvents, monomers and additives in materials (including stabilizers, antioxidants, plasticizers, colorants), and the like. Since leachables are harmful to the safety of the human body in a short or long term while the medical device is functioning as a medical instrument, it is necessary to evaluate the risk of the leachables of the medical device.
Although the leachables studied most effectively represented the amount of user contact during the actual application. However, due to practical challenges (including diversity and complexity of clinical access and methods of use, ethical issues with clinical specimen collection, etc.), leachables have been difficult to study in a true sense with most instruments, and it is desirable in some cases to replace leachables by leaching tests. The analysis methods currently used for leachables and leachables content are primarily known quantitation, which can leak out potential leachables, and a need exists for an analysis technique that does not know the leachables and leachables content.
Disclosure of Invention
In order to solve the technical problems, the invention provides a method for detecting the content of volatile and semi-volatile unknown substances in a medical apparatus leaching solution, which has the following technical scheme:
the method for detecting the content of volatile and semi-volatile unknown substances in the medical apparatus leaching liquor is characterized by comprising the following steps:
1) Preparing acetophenone series standard working solution, dimethyl phthalate series standard working solution, di-tert-butyl-4-methylphenol series standard working solution and alternative standard mixed solution, wherein the concentration gradients of the acetophenone, dimethyl phthalate and di-tert-butyl-4-methylphenol series standard working solution are set to be 0.5 mu g/mL-10 mu g/mL, and the dimethyl phthalate is added into the alternative standard mixed solution;
2) Respectively analyzing the acetophenone, dimethyl phthalate and di-tert-butyl-4-methylphenol series standard working solution by using a gas chromatography-mass spectrometer by adopting an internal standard method, carrying out linear regression analysis on the concentration and the peak area of the acetophenone series standard working solution to obtain an acetophenone linear regression equation and establish an acetophenone calibration curve, carrying out linear regression analysis on the concentration and the peak area of the dimethyl phthalate series standard working solution to obtain a dimethyl phthalate linear regression equation and establish a dimethyl phthalate calibration curve, and carrying out linear regression analysis on the concentration and the peak area of the di-tert-butyl-4-methylphenol series standard working solution to obtain a di-tert-butyl-4-methylphenol linear regression equation and establish a di-tert-butyl-4-methylphenol calibration curve;
3) Detecting the substitute standard substance mixed solution by using a gas chromatography-mass spectrometer to obtain the retention time of the substitute standard substance and the peak area of the substitute standard substance;
4) Calculating the relative response factor of the substitute standard substance in the substitute standard substance mixed solution relative to dimethyl phthalate, wherein the calculation formula is as follows:
in the formula:
RRF: relative response factor
A S : peak area of surrogate standard;
C S : surrogate standard concentrations;
A DMP : substituting the peak area of dimethyl phthalate in the standard substance mixed solution;
C DMP : replacing the concentration of dimethyl phthalate in the standard substance mixed solution;
5) Leaching medical instruments by using a leaching solvent to obtain a leaching solution, and detecting the leaching solution by using a gas chromatography-mass spectrometer by using an internal standard method to obtain the unknown substance retention time of the leaching solution and the unknown substance peak area of the leaching solution;
6) Respectively comparing the retention time of the unknown substances of the leaching liquor obtained in the step 5) with the retention time of acetophenone, dimethyl phthalate, di-tert-butyl-4-methylphenol and the retention time of the substitute standard substances obtained in the step 3) obtained in the step 2);
if the retention time of the unknown substances in the leaching liquor is similar to that of acetophenone, substituting the peak area of the unknown substances in the leaching liquor obtained in the step 5) into the acetophenone linear regression equation obtained in the step 2), and calculating to obtain the concentration of the unknown substances;
if the retention time of the unknown substances in the leaching liquor is similar to that of dimethyl phthalate, substituting the peak area of the unknown substances in the leaching liquor obtained in the step 5) into the linear regression equation of dimethyl phthalate obtained in the step 2), and calculating to obtain the concentration of the unknown substances;
if the retention time of the unknown substances of the leaching liquor is similar to that of the di-tert-butyl-4-methylphenol, substituting the peak area of the unknown substances of the leaching liquor obtained in the step 5) into the linear regression equation of the di-tert-butyl-4-methylphenol obtained in the step 2) to calculate the concentration of the unknown substances;
if the retention time of the unknown substances in the leaching liquor is similar to that of the substitute standard substances, substituting the peak area of the unknown substances in the leaching liquor obtained in the step 5) into the following calculation formula, and converting to obtain the relative peak area of the unknown substances:
in the formula:
A m : unknown relative peak area;
A n : peak area of unknown substance;
and (2) RRF: a relative response factor;
substituting the relative peak area of the unknown substance into the linear regression equation of the dimethyl phthalate obtained in the step 2), and calculating to obtain the concentration of the unknown substance;
if the retention time of the unknown substances in the leaching liquor is different from the retention time of acetophenone, the retention time of dimethyl phthalate, the retention time of di-tert-butyl-4-methylphenol and the retention time of the substitute standard substances, directly substituting the peak area of the unknown substances in the leaching liquor obtained in the step 5) into a regression equation of the retention time of the dimethyl phthalate obtained in the step 2), and calculating to obtain the concentration of the unknown substances;
7) Calculating the content of the unknown substances in the medical instrument product, wherein the calculation formula is as follows:
in the formula:
C is prepared from : unknown substance concentration;
V lifting device : volume of leach liquor;
n device for cleaning the skin : number of medical instruments leached.
Specifically, the concentrations of the acetophenone series standard working solution in the step 1) are respectively 0.5 mu g/mL, 1 mu g/mL, 2 mu g/mL, 5 mu g/mL and 10 mu g/mL, the concentrations of the dimethyl phthalate series standard working solution are respectively 0.5 mu g/mL, 1 mu g/mL, 2 mu g/mL, 5 mu g/mL and 10 mu g/mL, and the concentrations of the di-tert-butyl-4-methylphenol series standard working solution are respectively 0.5 mu g/mL, 1 mu g/mL, 2 mu g/mL, 5 mu g/mL and 10 mu g/mL.
Specifically, the mixed solution of the substitute standard in the step 1) further comprises N, N-dimethylpropionamide, caproic acid, 1-methyl-2-pyrrolidone, benzothiazole, 2-ethylhexyl acrylate, methyl benzoylformate, diphenyl ether, tetradecyl cycloheptasiloxane, diphenylamine, p-toluenesulfonamide, laurolactam, N-octadecane, butyl palmitate, tributyl acetylcitrate, and di (2-ethylhexyl) phthalate.
Specifically, the leaching liquor in step 5) is obtained by leaching the medical device product with the leaching solvent, and the leaching solvent is one or more of isopropanol, ethanol and water.
Specifically, when the extraction solvent is water or water and ethanol in equal proportion, the extraction solution is subjected to two liquid-liquid extractions by using dichloromethane, and organic phases obtained by the two liquid-liquid extractions are combined to be used as an extraction solution sample to be detected.
Specifically, in the step 2) and the step 5), the internal standard substance used by the internal standard method comprises 2-fluorobiphenyl and acenaphthene-d 10.
Specifically, in the step 2), the step 3) and the step 5), the gas chromatography conditions of the gas chromatography-mass spectrometer are as follows:
a chromatographic column: DB-1 capillary column 30m × 0.25mm × 0.25 μm; sample inlet temperature: at 210 ℃; sample introduction volume: 1.0 μ L; the flow splitting ratio is 5; the flow rate is 1.0mL/min; carrier gas: helium with purity of more than 99.999%; programmed temperature 40 ℃ for 1min, temperature at 10 ℃/min to 300 ℃, column maintenance for 10 min: DB-1 capillary column 30m × 0.25mm × 0.25 μm; sample inlet temperature: at 210 ℃; sample introduction volume: 1.0 μ L; the flow splitting ratio is 5; the flow rate is 1.0mL/min; carrier gas: helium with purity > 99.999%; the programmed temperature is maintained at 40 ℃ for 1min, and the temperature is raised to 300 ℃ at a speed of 10 ℃/min and maintained for 10min.
Specifically, in the step 5), the high-resolution mass spectrometry conditions of the gas chromatography-mass spectrometer are as follows:
the ion source is an electrospray ion source; transmission line temperature 280 ℃, ionization mode: electron bombardment; the ion source temperature is 230 ℃; quadrupole rod temperature: 150 ℃; ionization energy: 70eV; solvent delay 4.5min; the monitoring mode is as follows: a full scan mode; the mass scanning range of the mass spectrum is 30-650 amu; the mass spectrogram of the gas chromatography-mass spectrometer is used for carrying out qualitative analysis on the unknown substance.
Compared with the prior art, the invention has the beneficial effects that:
1. the invention can semi-quantitatively or quantitatively determine volatile and semi-volatile unknown substances in the medical apparatus leaching solution, makes up for the phenomenon of leachable substances omission caused by only paying attention to quantitative results of volatile and semi-volatile known substances, adopts a gas chromatography-high resolution mass spectrometer for analysis, and also reduces the cost for developing, verifying and testing various volatile and semi-volatile known substances.
2. Compared with the prior art, the content of volatile and semi-volatile unknown substances in the leaching liquor of the medical instrument product is measured by adopting a gas chromatography high-resolution mass spectrometer, the volatile and semi-volatile compounds which are released by the medical instrument product and are higher than an analysis threshold value can be identified, a compound dimethyl phthalate with relatively moderate response factors is selected as a calibration reference in all known substitute compounds, and semi-quantitative analysis is carried out on the compounds with structures, properties and retention time which are dissimilar to those of substitute standard substances in the leaching liquor sample solution, so that the accuracy and the comprehensiveness of detection are improved.
Drawings
FIG. 1 is a total ion flow diagram of dimethyl phthalate, acetophenone and di-tert-butyl-4-methylphenol and an internal standard in the present invention;
FIG. 2 is a schematic diagram of a calibration curve for dimethyl phthalate in accordance with the present invention;
FIG. 3 is a schematic diagram of an acetophenone calibration curve according to the present invention;
FIG. 4 is a graphical representation of a calibration curve for di-tert-butyl-4-methylphenol according to the present invention;
FIG. 5 is a gas chromatogram of 16 alternative compounds of the invention;
FIG. 6 is a gas chromatogram of a blank solution under water leaching conditions in accordance with the present invention;
FIG. 7 is a gas chromatogram of sample solution 1 (TW-1) under water leaching conditions in accordance with the present invention;
FIG. 8 is a gas chromatogram of sample solution 2 (TW-2) under water leaching conditions in accordance with the present invention;
FIG. 9 is a gas chromatogram of sample solution 3 (TW-3) under water leaching conditions in accordance with the present invention;
FIG. 10 is a gas chromatogram of a blank solution under ethanol leaching conditions in accordance with the present invention;
FIG. 11 is a gas chromatogram of sample solution 1 (TE-1) under the ethanol leaching conditions in the present invention;
FIG. 12 is a gas chromatogram of sample solution 2 (TE-2) under ethanol leaching conditions in the present invention;
FIG. 13 is a gas chromatogram of sample solution 3 (TE-3) under ethanol leaching conditions in the present invention;
FIG. 14 is a gas chromatogram of a blank solution under isopropanol leaching conditions in accordance with the invention;
FIG. 15 is a gas chromatogram of sample solution 1 (TP-1) under the conditions of isopropanol leaching in the present invention;
FIG. 16 is a gas chromatogram of sample solution 2 (TP-2) under the isopropanol leaching conditions in the present invention;
FIG. 17 is a gas chromatogram of sample solution 3 (TP-3) under the conditions of isopropanol leaching in the present invention;
FIG. 18 is a mass spectrum of the unknown at 5.53min of sample solution 3 (TP-3) under the conditions of isopropanol leaching in the present invention;
FIG. 19 is a mass spectrum of triisopropyl borate under chromatographic conditions according to the present invention;
FIG. 20 is a mass spectrum of the unknown at 10.63min of sample solution 3 (TP-3) under the isopropanol leaching condition in the present invention;
FIG. 21 is a mass spectrum of laurolactam under chromatographic conditions according to the present invention;
FIG. 22 is a mass spectrum of the unknown at 15.88min of sample solution 3 (TP-3) under the conditions of isopropanol leaching in the present invention;
FIG. 23 is a mass spectrum of n-hexadecane under chromatographic conditions according to the invention;
FIG. 24 is a mass spectrum of the unknown at 18.04min of sample solution 3 (TP-3) under the conditions of isopropanol leaching in the present invention;
FIG. 25 is a mass spectrum of n-octadecane under chromatographic conditions according to the invention;
FIG. 26 is a mass spectrum of the unknown at 26.08min of sample solution 3 (TP-3) under the isopropanol leaching condition in the present invention;
FIG. 27 is a mass spectrum of (Z) -erucamide under chromatographic conditions according to the invention;
FIG. 28 is a mass spectrum of the unknown at 27.30min of sample solution 3 (TP-3) under the conditions of isopropanol leaching in the present invention;
FIG. 29 is a mass spectrum of a siloxane related compound under chromatographic conditions in accordance with the present invention;
FIG. 30 is a mass spectrum of an unknown at 32.13min of sample solution 3 (TP-3) under the conditions of isopropanol leaching in the present invention;
FIG. 31 is a mass spectrum of antioxidant 168 under chromatographic conditions in accordance with the present invention.
Detailed Description
The technical solutions of the present invention will be described clearly and completely below, and it should be apparent that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be obtained by a person skilled in the art without making any creative effort based on the embodiments in the present invention, belong to the protection scope of the present invention. Materials, instruments, reagents and the like used in the following examples are commercially available unless otherwise specified.
Example one
In the embodiment, a medical instrument nasal cannula is selected to detect the content of volatile and semi-volatile unknown substances in the nasal cannula leaching liquor.
1. Solution preparation
1.1 preparing a mixed solution of the substitute standard substance:
weighing about 20mg of each substitute standard substance in table 1 in the same 20mL volumetric flask, fixing the volume to the scale mark by using dichloromethane, precisely moving 1mL in the 20mL volumetric flask after uniformly mixing, fixing the volume to the scale mark by using dichloromethane, uniformly mixing, precisely moving 1mL in the 10mL volumetric flask after uniformly mixing, fixing the volume to the scale mark by using dichloromethane, and uniformly mixing to serve as a substitute standard substance mixed solution.
Table 1:16 substitute standard products
| Name of substitute standard | Abbreviations | CAS# |
| N, N-dimethylpropionamide | DMPA | 758-96-3 |
| Hexanoic acid | HA | 142-62-1 |
| 1-methyl-2-pyrrolidone | NMP | 872-50-4 |
| Benzothiazole compounds | BT | 95-16-9 |
| 2-ethylhexyl acrylate | EHA | 103-11-7 |
| Benzoic acid methyl ester | MOPA | 15206-55-0 |
| Diphenyl ether | DPE | 101-84-8 |
| Decamethylcycloheptasiloxane | TDCHS | 107-50-6 |
| Diphenylamine | DPA | 122-39-4 |
| Para toluene sulfonamide | TS | 70-55-3 |
| Laurolactam | DDL | 947-04-6 |
| N-octadecane | OD | 593-45-3 |
| Hexadecanoic acid butyl ester | BP | 111-06-8 |
| Acetyl tributyl citrate | TBA | 77-90-7 |
| Di (2-ethylhexyl) phthalate | DEHP | 117-81-7 |
| Phthalic acid dimethyl ester | DMP | 131-11-3 |
| Name of substitute standard | Abbreviations | CAS# |
| N, N-dimethylpropionamide | DMPA | 758-96-3 |
1.2 internal standard solution preparation:
1.2.1 internal standard solution: respectively weighing about 20mg of 2-fluorobiphenyl and acenaphthene-d 10 in the same 20mL volumetric flask, and metering the volume to a scale mark by using dichloromethane, wherein the scale mark is SS-ISTD;
1.2.2 internal standard intermediate stock solutions: precisely transferring 1mL of internal standard solution into a 5mL volumetric flask, and metering the volume to a scale mark by using dichloromethane, wherein the mark is ISS-ISTD # 1;
1.2.3 internal standard working solution: precisely transferring 1mL of internal standard intermediate stock solution into a 100mL volumetric flask, and carrying out constant volume to a scale mark with dichloromethane, wherein the mark is ISTD.
1.3 preparing acetophenone, dimethyl phthalate and di-tert-butyl-4-methylphenol series standard working solutions:
1.3.1 dimethyl phthalate standard solution: weighing 20mg dimethyl phthalate (DMP) in a 20mL volumetric flask, metering volume to a scale mark by using ISTD, uniformly mixing, and marking as SS # 1;
standard intermediate stock solution 1: precisely transferring 1mL of standard solution into a 10mL volumetric flask, carrying out constant volume to a scale mark by using ISTD, uniformly mixing, and marking as ISS # 1;
standard intermediate stock solution 2: precisely transferring 1mL of the standard solution into a 20mL volumetric flask, carrying out constant volume metering to a scale mark by using the ISTD, and uniformly mixing, wherein the mark is ISS # 2.
1.3.2 dimethyl phthalate series standard working solutions:
the series of standard working solutions were labeled L-1, L-2, L-3, L-4, and L-5, where:
the formulation of L-4 (5. Mu.g/ml) was: accurately weighing 1ml of ISS # 2 in a 10ml volumetric flask, metering to a scale mark by using ISTD, and uniformly mixing;
the formulation of L-5 (10. Mu.g/ml) was: accurately weighing 1ml of ISS # 1 in a 10ml volumetric flask, metering to a scale mark by using ISTD, and uniformly mixing;
the formulation procedure for L-1 (0.5. Mu.g/ml) was: precisely measuring 1ml of L-4 in a 10ml volumetric flask, metering to a scale mark by using ISTD, and uniformly mixing;
the formulation of L-2 (1. Mu.g/ml) was: precisely measuring 1ml of L-5 in a 10ml volumetric flask, metering to a scale mark by using ISTD, and uniformly mixing;
the preparation process of L-3 (2. Mu.g/ml) is as follows: precisely measuring 1ml of L-5 in a 5ml volumetric flask, metering to a scale mark by using ISTD, and uniformly mixing;
1.3.3 preparing acetophenone series standard substance working solution and di-tert-butyl-4-methylphenol series standard substance working solution according to the same steps of 1.3.1 and 1.3.2.
1.4 extraction liquid sample acquisition:
1.4.1 isopropanol leach liquor: taking 10mL of isopropanol-leached solution, adding 100 mu L of internal standard intermediate stock solution, preparing 3 parts in parallel, respectively marking as TP-1, TP-2 and TP-3, and waiting for machine analysis;
1.4.2 water extract: taking 10mL of solution after water extraction, adding 100 mu L of internal standard intermediate storage solution, adding 5mL of dichloromethane for liquid-liquid extraction, deflating, standing, and transferring the lower-layer organic phase into another sample bottle after layering; adding 5mL of dichloromethane, repeatedly extracting for the second time, and combining organic phases; preparing 3 parts in parallel, marking as TW-1, TW-2 and TW-3 respectively, and waiting for machine analysis;
1.4.3 ethanol leaching solution: taking 10mL of ethanol leached solution, and adding 100 mu L of internal standard intermediate stock solution; 3 parts are prepared in parallel and marked as TE-1, TE-2 and TE-3 respectively, and are ready to be analyzed on a computer.
1.5 preparing blank leaching liquor:
1.5.1 isopropanol blank leach liquor: taking 10mL of isopropanol, and adding 100 mu L of internal standard intermediate stock solution;
1.5.2 water blank leach liquor: taking 10mL of water, adding 100 mu L of internal standard intermediate storage solution, adding 5mL of dichloromethane for liquid-liquid extraction, deflating, standing, and transferring the lower-layer organic phase into another sample bottle after layering; adding 5mL of dichloromethane, repeatedly extracting for the second time, and combining organic phases;
1.5.3 ethanol blank leaching solution: 10mL of ethanol was taken and 100. Mu.L of internal standard intermediate stock solution was added. As shown in figure 1, the total ion flow diagram of dimethyl phthalate, acetophenone, di-tert-butyl-4-methylphenol and an internal standard substance is shown.
2. Analysis conditions of high performance gas chromatography-mass spectrometer
2.1 high performance gas chromatography conditions:
a chromatographic column: DB-1 capillary column 30m × 0.25mm × 0.25 μm; sample inlet temperature: at 210 ℃; sample introduction volume: 1.0 μ L; the flow splitting ratio is 5; the flow rate is 1.0mL/min; carrier gas: helium with purity > 99.999%; maintaining the program temperature at 40 deg.C for 1min, heating to 300 deg.C at 10 deg.C/min, and maintaining for 10min;
2.2 Mass Spectrometry conditions:
3. analyzing and detecting by adopting high-efficiency gas chromatography-mass spectrometry
3.1 detecting the mixed solution of the substitute standard substance, isopropanol leaching liquor, water leaching liquor, ethanol leaching liquor, acetophenone series standard working solution, dimethyl phthalate series standard working solution, di-tert-butyl-4-methylphenol series standard working solution, isopropanol blank leaching liquor, water blank leaching liquor and ethanol blank leaching liquor by using a high performance gas chromatography-mass spectrometer, and recording a chromatogram;
3.2 the analysis results are shown in FIGS. 5-17, and the peak area of each alternative standard in the alternative standard mixed solution, the retention time of the unknown substance in each leaching solution and the peak area of the unknown substance are obtained.
4. Data processing
4.1, performing integration treatment, recording a chromatogram, and performing linear regression analysis on concentration data of acetophenone, dimethyl phthalate and di-tert-butyl-4-methylphenol in the series of standard working solutions and a chromatogram peak area to obtain linear regression equations of acetophenone, dimethyl phthalate and di-tert-butyl-4-methylphenol, wherein the linear regression equations are shown in fig. 2-4, and a standard curve is prepared according to the linear regression equations, and are shown in the following table 2:
table 2: linear regression equation of acetophenone, dimethyl phthalate and di-tert-butyl-4-methylphenol series standard working solution
| Series of standard working solutions | Linear regression equation (y: peak area x: concentration) |
| Acetophenone | y＝0.493252x-0.059153 |
| Phthalic acid dimethyl ester | y＝0.541279x-0.027891 |
| Di-tert-butyl-4-methylphenol | y＝0.889615x-0.068866 |
4.2 the relative response factors of the 16 alternative standards relative to dimethyl phthalate were calculated according to the following formula:
in the formula:
RRF: a relative response factor;
A S : alternative standard peak area;
C S : surrogate standard concentrations;
A DMP : substituting the peak area of dimethyl phthalate in the standard substance mixed solution;
C DMP : replacing the concentration of dimethyl phthalate in the standard substance mixed solution;
the results are shown in table 3 below:
table 3: relative response factors of 16 substitute standards relative to dimethyl phthalate
The relative response factor is used for representing the relative relation between the content of the dimethyl phthalate in the substitute mixed solution and the substitute mixed solution.
5. Calculating the concentration of unknown substance
5.1, identifying the retention time of the unknown substances in the leaching liquor, substituting the peak areas of the unknown substances into a dimethyl phthalate linear regression equation in a corresponding mode when the retention time of the unknown substances is similar to the retention time of dimethyl phthalate so as to calculate the concentration of the unknown substances, and directly combining a high-resolution mass spectrogram to qualitatively analyze the unknown substances because the retention time of the unknown substances is similar to the retention time of the dimethyl phthalate;
5.2, identifying the retention time of the unknown substance in the leaching liquor, substituting the peak area of the unknown substance into an acetophenone linear regression equation under a corresponding mode when the retention time of the unknown substance is similar to that of acetophenone, so as to calculate the concentration of the unknown substance, and directly combining a high-resolution mass spectrogram to perform qualitative analysis on the unknown substance because the retention time of the unknown substance is similar to that of acetophenone;
5.3 identifying the retention time of the unknown substance in the leaching liquor, substituting the peak area of the unknown substance into a linear regression equation of the di-tert-butyl-4-methylphenol in a corresponding mode when the retention time of the unknown substance is similar to that of the di-tert-butyl-4-methylphenol, thereby calculating the concentration of the unknown substance, and directly combining a high-resolution mass spectrogram to carry out qualitative analysis on the unknown substance because the retention time of the unknown substance is similar to that of the di-tert-butyl-4-methylphenol; 5.4 when the retention time of the unknown substance is similar to that of a certain substitute standard, firstly calculating the peak area of the unknown substance relative to the dimethyl phthalate series standard working solution according to the following formula, namely the relative peak area of the unknown substance:
in the formula:
A m : unknown relative peak area;
A n : peak area of unknown substance;
RRF: a relative response factor;
substituting the relative peak area of the unknown substance obtained by calculation into a dimethyl phthalate linear regression equation to calculate the concentration of the unknown substance;
5.5 when the retention time of the unknown substances in the leaching liquor is different from that of acetophenone, dimethyl phthalate, di-tert-butyl-4-methylphenol and the substitute standard, directly substituting the peak area of the unknown substances in the leaching liquor into a dimethyl phthalate linear regression equation to calculate the concentration of the unknown substances, and then carrying out semi-quantitative analysis on the unknown substances in the leaching liquor.
6. Calculating the content of unknown substance
6.1 according to the calculated concentration of the unknown substance, calculating the content of the unknown substance in the medical appliance product, wherein the calculation formula is as follows:
in the formula:
C is prepared from : unknown substance concentration;
V lifting device : volume of leach liquor;
n device for cleaning the skin : the number of medical instruments to be extracted;
6.2 taking the extracted solution TP-3 of the isopropanol sample as an example, as shown in FIG. 17, it is the gas chromatography-mass spectrometry analysis result of the extracted solution TP-3 of the isopropanol sample, 7 unknowns are selected for a representative quantitative method description, the mass spectrograms of the 7 unknowns and 7 standards or substitute standards are shown in FIGS. 18-31, and the specific identification and quantitative results are shown in Table 4 below:
table 4: identification and quantification results of 7 unknowns in isopropanol sample leaching solution TP-3
The method for detecting the content of the volatile and semi-volatile unknown substances in the medical apparatus leaching solution provided by the embodiment can identify all volatile and semi-volatile compounds which are released by the medical apparatus product and are higher than an analysis threshold value, makes up for the defect of only carrying out quantitative analysis on the known substances, improves the safety of the medical apparatus product in the using process, and is also beneficial to reducing the cost of manpower and material resources of a medical apparatus manufacturer in the production, development and verification processes.
In summary, the above-mentioned embodiments are merely preferred embodiments of the present invention, and are not intended to limit the scope of the present invention, and any modifications, equivalents, improvements, etc. made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (5)
1. The method for detecting the content of volatile and semi-volatile unknown substances in the medical apparatus leaching liquor is characterized by comprising the following steps of:
1) Preparing acetophenone series standard working solution, dimethyl phthalate series standard working solution, di-tert-butyl-4-methylphenol series standard working solution and alternative standard mixed solution, wherein the concentration gradients of the acetophenone, dimethyl phthalate and di-tert-butyl-4-methylphenol series standard working solution are set to be 0.5 mu g/mL-10 mu g/mL, and the dimethyl phthalate is added into the alternative standard mixed solution;
2) Respectively analyzing the acetophenone, dimethyl phthalate and di-tert-butyl-4-methylphenol series standard working solution by using a gas chromatography-mass spectrometer by adopting an internal standard method, carrying out linear regression analysis on the concentration and the peak area of the acetophenone series standard working solution to obtain an acetophenone linear regression equation and establish an acetophenone calibration curve, carrying out linear regression analysis on the concentration and the peak area of the dimethyl phthalate series standard working solution to obtain a dimethyl phthalate linear regression equation and establish a dimethyl phthalate calibration curve, and carrying out linear regression analysis on the concentration and the peak area of the di-tert-butyl-4-methylphenol series standard working solution to obtain a di-tert-butyl-4-methylphenol linear regression equation and establish a di-tert-butyl-4-methylphenol calibration curve;
3) Detecting the substitute standard substance mixed solution by using a gas chromatography-mass spectrometer to obtain the retention time of the substitute standard substance and the peak area of the substitute standard substance;
4) Calculating the relative response factor of the substitute standard substance in the substitute standard substance mixed solution relative to dimethyl phthalate, wherein the calculation formula is as follows:
in the formula:
RRF: relative response factor
A S : alternative standard peak area;
C S : surrogate standard concentrations;
C DMP : replacing the concentration of dimethyl phthalate in the standard substance mixed solution;
5) Leaching medical instruments by using a leaching solvent to obtain a leaching solution, and detecting the leaching solution by using a gas chromatography-mass spectrometer by using an internal standard method to obtain the unknown substance retention time of the leaching solution and the unknown substance peak area of the leaching solution;
6) Respectively comparing the retention time of the unknown substances of the leaching liquor obtained in the step 5) with the retention time of acetophenone, dimethyl phthalate, di-tert-butyl-4-methylphenol and the retention time of the substitute standard substances obtained in the step 3) obtained in the step 2);
if the retention time of the unknown substances in the leaching liquor is similar to that of acetophenone, substituting the peak area of the unknown substances in the leaching liquor obtained in the step 5) into the acetophenone linear regression equation obtained in the step 2), and calculating to obtain the concentration of the unknown substances;
if the retention time of the unknown substances of the leaching liquor is similar to that of dimethyl phthalate, substituting the peak area of the unknown substances of the leaching liquor obtained in the step 5) into the linear regression equation of dimethyl phthalate obtained in the step 2), and calculating to obtain the concentration of the unknown substances;
if the retention time of the unknown substances of the leaching liquor is similar to that of the di-tert-butyl-4-methylphenol, substituting the peak area of the unknown substances of the leaching liquor obtained in the step 5) into the linear regression equation of the di-tert-butyl-4-methylphenol obtained in the step 2), and calculating to obtain the concentration of the unknown substances;
if the retention time of the unknown substances in the leaching liquor is similar to that of the substitute standard substances, substituting the peak area of the unknown substances in the leaching liquor obtained in the step 5) into the following calculation formula, and converting to obtain the relative peak area of the unknown substances:
in the formula:
A m : the relative peak area of the unknown object;
A n : peak area of unknown substance;
and (2) RRF: a relative response factor;
substituting the relative peak area of the unknown substance into the linear regression equation of the dimethyl phthalate obtained in the step 2), and calculating to obtain the concentration of the unknown substance;
if the retention time of the unknown substances in the leaching liquor is different from the retention time of acetophenone, the retention time of dimethyl phthalate, the retention time of di-tert-butyl-4-methylphenol and the retention time of substitute standard substances, directly substituting the peak area of the unknown substances in the leaching liquor obtained in the step 5) into a regression equation of the retention time of the dimethyl phthalate obtained in the step 2), and calculating to obtain the concentration of the unknown substances;
7) Calculating the content of the unknown substances in the medical appliance product, wherein the calculation formula is as follows:
in the formula:
C is prepared from : unknown concentration;
V lifting device : volume of leach liquor；
n Device for cleaning the skin : the number of medical instruments to be extracted;
the mixed solution of the substitute standard substance in the step 1) also comprises N, N-dimethylpropionamide, caproic acid, 1-methyl-2-pyrrolidone, benzothiazole, 2-ethylhexyl acrylate, methyl benzoylformate, diphenyl ether, tetradecyl cyclo-heptasiloxane, diphenylamine, p-toluenesulfonamide, laurolactam, N-octadecane, butyl hexadecanoate, acetyl tributyl citrate and di (2-ethylhexyl) phthalate;
in the step 2), the step 3) and the step 5), the gas chromatography conditions of the gas chromatography-mass spectrometer are as follows:
and (3) chromatographic column: DB-1 capillary column 30m × 0.25mm × 0.25 μm; sample inlet temperature: at 210 ℃; sample injection volume: 1.0 Mu L; the flow splitting ratio is 5; the flow rate is 1.0mL/min; carrier gas: helium with purity > 99.999%; temperature rising procedure: maintaining at 40 deg.C for 1min, heating to 300 deg.C at 10 deg.C/min, and maintaining for 10min;
the mass spectrum conditions of the gas chromatography-mass spectrometer are as follows:
transmission line temperature 280 ℃, ionization mode: electron bombardment; ion source temperature: 230 ℃; temperature of the quadrupole rods: 150 ℃; ionization energy: 70eV; solvent delay 4.5min; the monitoring mode is as follows: a full scan mode; mass spectrum mass scanning range is 30 to 650amu; the mass spectrogram of the gas chromatography-mass spectrometer is used for carrying out qualitative analysis on the unknown substance.
2. The detection method according to claim 1, wherein the concentrations of the acetophenone series standard working solutions in step 1) are 0.5 μ g/mL, 1 μ g/mL, 2 μ g/mL, 5 μ g/mL and 10 μ g/mL, respectively, the concentrations of the dimethyl phthalate series standard working solutions are 0.5 μ g/mL, 1 μ g/mL, 2 μ g/mL, 5 μ g/mL and 10 μ g/mL, respectively, and the concentrations of the di-tert-butyl-4-methylphenol series standard working solutions are 0.5 μ g/mL, 1 μ g/mL, 2 μ g/mL, 5 μ g/mL and 10 μ g/mL, respectively.
3. The method for detecting according to claim 1, wherein the leaching solution in step 5) is obtained by leaching the medical device product with the leaching solvent, and the leaching solvent is selected from one or more of isopropyl alcohol, ethanol, and water.
4. The detection method according to claim 3, wherein when the extraction solvent is selected from water or water and ethanol in equal proportion, the extraction solution is subjected to two liquid-liquid extractions with dichloromethane, and organic phases obtained by the two liquid-liquid extractions are combined to be used as a sample of the extraction solution to be detected.
5. The detection method according to claim 1, wherein in the step 2) and the step 5), the internal standard substance used in the internal standard method comprises 2-fluorobiphenyl and acenaphthene-d 10.
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