CN117783037A - Method for rapidly confirming main components of filler by utilizing infrared spectrometry - Google Patents
Method for rapidly confirming main components of filler by utilizing infrared spectrometry Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 43
- 239000000945 filler Substances 0.000 title abstract description 43
- 238000004566 IR spectroscopy Methods 0.000 title description 12
- 239000002245 particle Substances 0.000 claims abstract description 139
- 238000004458 analytical method Methods 0.000 claims abstract description 19
- 238000002329 infrared spectrum Methods 0.000 claims abstract description 19
- 238000001514 detection method Methods 0.000 claims abstract description 15
- 230000005540 biological transmission Effects 0.000 claims description 29
- KRHYYFGTRYWZRS-UHFFFAOYSA-M Fluoride anion Chemical compound [F-] KRHYYFGTRYWZRS-UHFFFAOYSA-M 0.000 claims description 28
- 238000012856 packing Methods 0.000 claims description 23
- 238000002156 mixing Methods 0.000 claims description 18
- 230000003287 optical effect Effects 0.000 claims description 17
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 claims description 12
- 239000003085 diluting agent Substances 0.000 claims description 10
- 239000000243 solution Substances 0.000 claims description 10
- OYLGJCQECKOTOL-UHFFFAOYSA-L barium fluoride Chemical compound [F-].[F-].[Ba+2] OYLGJCQECKOTOL-UHFFFAOYSA-L 0.000 claims description 9
- 229910001632 barium fluoride Inorganic materials 0.000 claims description 9
- 238000007865 diluting Methods 0.000 claims description 9
- 239000004005 microsphere Substances 0.000 claims description 8
- 238000010790 dilution Methods 0.000 claims description 7
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- 229910016036 BaF 2 Inorganic materials 0.000 claims description 6
- 238000007906 compression Methods 0.000 claims description 5
- 229910004261 CaF 2 Inorganic materials 0.000 claims description 3
- WUKWITHWXAAZEY-UHFFFAOYSA-L calcium difluoride Chemical compound [F-].[F-].[Ca+2] WUKWITHWXAAZEY-UHFFFAOYSA-L 0.000 claims description 3
- 229910001634 calcium fluoride Inorganic materials 0.000 claims description 3
- ORUIBWPALBXDOA-UHFFFAOYSA-L magnesium fluoride Chemical compound [F-].[F-].[Mg+2] ORUIBWPALBXDOA-UHFFFAOYSA-L 0.000 claims description 3
- 229910001635 magnesium fluoride Inorganic materials 0.000 claims description 3
- 230000008569 process Effects 0.000 abstract description 8
- 239000000523 sample Substances 0.000 description 64
- 238000005102 attenuated total reflection Methods 0.000 description 16
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- 238000004587 chromatography analysis Methods 0.000 description 9
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- 125000004029 hydroxymethyl group Chemical group [H]OC([H])([H])* 0.000 description 3
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- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 2
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Abstract
The invention relates to a method for rapidly confirming main components of a filler by utilizing an infrared spectrum method, which is an analysis and detection method for main components of microspherical particles of a chromatographic column filler type. The infrared spectrogram obtained by the method has good matching degree with the standard spectrogram, the detection process is quick and efficient, multiple attempts are not needed, and the main components of the microspherical particles can be accurately detected.
Description
Technical Field
The invention belongs to the field of biological pharmacy, and relates to a method for rapidly confirming main components of a filler by utilizing an infrared spectrometry, wherein the filler is chromatographic column filler used in the processes of chromatography, ion exchange and the like in the production process of biological medicine products.
Background
The infrared spectrum technology is widely applied to the fields of chemical analysis, food industry, material science, environmental science and the like, and has the characteristics of high sensitivity, high speed, low cost, nondestructive analysis, online real-time analysis and the like. In the prior art, infrared spectroscopy has been successfully applied to geology (performing mineral analysis of geological samples, identifying mineral components of rock), environmental pollutant analysis (e.g., analysis of microplastic in the environment), water quality pollutant analysis (e.g., analysis of microplastic in drinking water, prediction of the concentration of nitrite and BOD in water), and the like (see, for example, patent documents 1 to 4).
On the other hand, in the field of biopharmaceuticals, particularly in the processes of chromatography, ion exchange and the like in the production process of biomedical products, the chromatography technology is an important drug analysis technology, almost runs through the whole process of drug research and development, and plays an indispensable role in links such as early target identification, lead molecular screening, process research and development, product quality research and control, impurity analysis and identification, pharmacological drug generation, clinical sample analysis and the like. As one of the most important devices in chromatography, the packing of a chromatographic column may be called the core of the chromatography, which is not only the basis for the chromatographic method, but its quality directly determines the resolution of the chromatographic column. Therefore, there is a general need in the art to detect the quality of chromatographic column packing by infrared spectroscopic techniques in order to confirm whether the packing has quality problems such as discoloration.
However, there are some problems specific to the column packing when the infrared spectroscopic technique is applied to the column packing. For example, unlike an analyte such as a contaminant, which is present in a low concentration, a column packing is generally stored in a particulate form in a specific solution in a high concentration, and if an infrared spectrometry is performed by taking a certain amount of the solution directly or after air-drying, the packing particles are aggregated together, resulting in indistinguishability and difficulty in the infrared spectrometry. In addition, unlike an irregularly shaped analyte such as a microplastic, the filler particles themselves have a high thickness and are easily moved in a spherical shape, and thus, have problems such as a large energy loss due to a long optical path, unstable test, and poor signals. Due to the problems, when the infrared spectrum technology is applied to chromatographic column packing, the infrared spectrum with better quality is difficult to obtain in a short time, and the main components of the infrared spectrum are affected to be identified and analyzed.
Therefore, there is an urgent need in the art to develop a specific analytical detection method for rapidly analyzing the main components of microspheroidal particles of the column packing type using infrared spectroscopic techniques.
Prior art literature
Patent literature:
patent document 1: chinese patent publication number CN115561200a
Patent document 2: chinese patent publication number CN116893156a
Patent document 3: chinese patent publication number CN113533239a
Patent document 4: chinese patent publication number CN117079735a
Disclosure of Invention
In view of the problems of the prior art, the present invention aims to provide a specific analysis and detection method for rapidly analyzing the main components of microspheroidal particles of column packing materials by using infrared spectroscopy.
In order to solve the above technical problems, through long-term practice and research, the present inventors found and proposed a specific analytical detection method for rapidly analyzing the main components of column packing type microspheroidal particles using infrared spectroscopic techniques. By adopting the method, the infrared spectrum of the spherical particles can be rapidly and stably obtained, and the main components of the particles can be obtained; further, when different kinds of spherical particles are mixed together, the particles from different sources can be rapidly distinguished; if the properties of part of the filler change during the use process to influence the production, the conditions of color change and the like of part of the filler are observed, and the changed particles can be rapidly positioned and analyzed.
Specifically, the invention provides an analysis and detection method for main components of microspheroidal particles, which comprises the following steps:
(1) Diluting the microspherical particle sample by a specified multiple by using a diluent, and uniformly mixing for later use;
(2) Diluting and uniformly mixing the microspherical particle sample, measuring a specified amount of diluted microspherical particle sample, dripping the diluted microspherical particle sample on a fluoride window, and placing the fluoride window at the center of a groove at the lower layer of the micro-compression dish;
(3) Covering the fluoride window sheet with another fluoride window sheet, screwing the fluoride window sheet with the microsphere particle sample to prepare a tabletting, and observing the tabletting microsphere particle sample by using an optical microscope and photographing;
(4) Observing the pressed microspherical particles by using a Fourier transform infrared microscope, taking a picture, and measuring an infrared transmission spectrogram of the specific microspherical particles;
(5) And (3) analyzing and comparing the obtained infrared transmission spectrogram with spectrograms in a commercial database by using software, so as to identify the main components of the sample.
In some preferred embodiments, the microspheroidal particles may be chromatographic column packing particles.
In some preferred embodiments, the microspheroidal particles may be agarose based column packing particles or hydroxymethyl acrylic polymer based column packing particles.
In some preferred embodiments, prior to the step (1), the microspheroidal particles stored in solution may be observed using an optical microscope.
In some preferred embodiments, prior to said step (1), the morphology, size, color of said microspheroidal particles stored in solution may be observed using an optical microscope.
In some preferred embodiments, the prescribed dilution factor in step (1) may be 30 to 100 times, preferably 40 to 70 times, more preferably 50 times.
In some preferred embodiments, the mixing in step (1) may be vortex mixing.
In some preferred embodiments, the fluoride window in step (2) may be barium fluoride BaF 2 Window pane, calcium fluoride CaF 2 Window or magnesium fluoride MgF 2 Window, preferably barium fluoride BaF 2 And a window sheet.
In some preferred embodiments, the prescribed amount of the diluted microspheroidal particle sample in step (2) may be from 0.5 to 2. Mu.L, preferably 1. Mu.L.
In some preferred embodiments, the optical microscopy observation in the step (3) may be observation of the morphology, size, and color of the sample of microspheroidal particles after tabletting using an optical microscope.
In some preferred embodiments, the fourier transform infrared microscope observation in the step (4) may be observation of the morphology and size of the sample of microspheroidal particles after tabletting by using a fourier transform infrared microscope.
In some preferred embodiments, the software used in step (5) may be KIA software (WileySoftware), the search method used for analysis and comparison may be a "first derivative Euclidean distance algorithm".
In some preferred embodiments, the commercial database used in step (5) may be a Sadtler infrared spectrum database.
The analysis and detection method can effectively distinguish and fix the chromatographic column packing microspherical particles, and the main components of the particles to be detected can be rapidly obtained by measuring the infrared transmission spectrogram of the chromatographic column packing microspherical particles by an infrared microscope.
Drawings
FIG. 1 is a schematic diagram of an infrared-transmissive sample.
Fig. 2 is an Optical Microscope (OM) image of a transmitted sample of agarose-based chromatographic column packing particles.
Figure 3 is an OM image of a transmitted sample of chromatographic column packing particles based on a methylol acrylic polymer.
Figure 4 is an OM image of agarose-based column packing particles after preparation on a silvered lens.
Figure 5 is an OM image of a hydroxymethyl acrylic polymer based chromatographic column filler particle after preparation on a silver coated lens.
FIG. 6 is an infrared spectrum of agarose-based chromatographic column packing particles in infrared reflectance, attenuated Total Reflectance (ATR) and transmission modes and their comparison to a database.
FIG. 7 is an infrared spectrum of a chromatographic column packing particle based on a methylol acrylic polymer in infrared reflection, ATR and transmission modes and its comparison with a database.
Detailed Description
The invention provides an analysis and detection method of main components of microspherical particles, which comprises the following steps:
(1) Diluting the microspherical particle sample by a specified multiple by using a diluent, and uniformly mixing for later use;
(2) Diluting and uniformly mixing the microspherical particle sample, measuring a specified amount of diluted microspherical particle sample, dripping the diluted microspherical particle sample on a fluoride window, and placing the fluoride window at the center of a groove at the lower layer of the micro-compression dish;
(3) Covering the fluoride window sheet with another fluoride window sheet, screwing the fluoride window sheet with the microsphere particle sample to prepare a tabletting, and observing the tabletting microsphere particle sample by using an optical microscope and photographing;
(4) Observing the pressed microspherical particles by using a Fourier transform infrared microscope, taking a picture, and measuring an infrared transmission spectrogram of the specific microspherical particles;
(5) And (3) analyzing and comparing the obtained infrared transmission spectrogram with spectrograms in a commercial database by using software, so as to identify the main components of the sample.
Step (1)
In the invention, the step (1) is a step of diluting the microspheroidal particle sample by a prescribed multiple with a diluent, and uniformly mixing for later use. The reason for this step (1) is the following new findings of the present inventors:
in the case of preparing a sample, if the sample of microspheroidal particles stored in the solution is directly prepared without dilution, more microspheroidal particles are aggregated within a specific range, and the microspheroidal particles cannot be distinguished from each other, so that the particles are overlapped and aggregated after the preparation of the infrared transmission sample and tabletting, and the infrared spectrogram of the specific particles cannot be acquired.
Accordingly, in view of the above new findings, the present inventors have improved the prior art method by diluting a sample of microspheroidal particles by a prescribed multiple and mixing them uniformly using a diluent at the time of sample preparation. Therefore, the problem of aggregation of microspherical particles can be effectively avoided, so that the particles can be clearly distinguished after the infrared transmission sample is prepared and pressed, and a high-quality infrared spectrogram of specific particles can be clearly acquired.
In some embodiments of step (1), the microspheroidal particles are chromatography column filler particles, more preferably agarose-based chromatography column filler particles or hydroxymethyl acrylic polymer-based chromatography column filler particles. Without wishing to be bound by any particular theory, since the chromatographic column packing particles, especially those based on agarose or methylol acrylic polymers, are susceptible to the above aggregation problems during sample preparation and are soft particles, the tabletting in step (3) below is easier to carry out than hard particles, and thus the method of the present invention is particularly suitable for solving the above aggregation problems and thus for efficiently identifying the main components thereof.
In some embodiments of step (1), prior to step (1), the microspheroidal particles stored in solution are observed using an optical microscope, in particular the morphology, size, colour of the microspheroidal particles. By this step, it is not only facilitated to more accurately determine the appropriate dilution factor in step (1), but also to select appropriate microspheroidal particles for subsequent infrared transmission spectrogram determinations.
In step (1), the diluent may be any kind of diluent commonly used in the infrared spectrum field as long as it can sufficiently dilute the microspheroidal particles without adversely affecting them. In some embodiments, the diluent is spectrally pure water, preferably ultrapure water.
In the step (1), the predetermined dilution ratio may be appropriately selected according to the need (for example, according to the material, morphology, size, and color of the microspheroidal particles), and is not particularly limited. In some embodiments, the prescribed dilution factor is, for example, 30 to 100 times, preferably 40 to 70 times, more preferably 50 times.
In step (1), the mixing may be performed by any method for mixing known in the art, and is not particularly limited as long as the method can uniformly mix the microspheroidal particles in the diluent. In some embodiments, the mixing is vortex mixing.
Step (2)
In the invention, the step (2) is a step of diluting and uniformly mixing the microspherical particle sample, measuring a specified amount of diluted microspherical particle sample, dripping the diluted microspherical particle sample on a fluoride window, and placing the fluoride window at the center of a groove at the lower layer of the micro-compression dish. The reason for this step (2) is the following new findings of the present inventors:
if the microspherical particles stored in the solution are directly dripped or moved to a conventional substrate such as a gold-plated filter film, a gold-plated lens or a silver-plated lens after being diluted, the microspherical particles are overlapped and aggregated together after air drying, the particles cannot be distinguished, and an infrared spectrogram of the specific particles cannot be acquired.
Accordingly, in view of the new findings described above, the present inventors have improved on the prior art methods by using fluoride window sheets instead of other conventional substrates such as gold-plated filters, gold-plated lenses, or silver-plated lenses in the preparation of samples. Therefore, the problem of aggregation of microspherical particles can be effectively avoided, so that the particles can be clearly distinguished after the infrared transmission sample is prepared and pressed, and a high-quality infrared spectrogram of specific particles can be clearly acquired.
Without wishing to be bound by any particular theory, it is believed that the use of fluoride window in step (2) is more desirable for use in microspheroidal particles stored in solution, probably because fluoride window is less prone to moisture than other conventional substrates such as gold-plated filters, gold-plated lenses or silver-plated lensesInfrared spectroscopic measurement of a particulate sample, particularly a sample of microspheroidal particles dispersed in a liquid after dilution with a diluent in step (1) above. Thus, in step (2), any fluoride window commonly used in the infrared spectroscopy field may be used as long as it is not easily wetted. In some embodiments, the fluoride window is barium fluoride BaF 2 Window pane, calcium fluoride CaF 2 Window or magnesium fluoride MgF 2 Window, preferably barium fluoride BaF 2 And a window sheet.
In the step (2), the predetermined amount of the diluted sample of the microspheroidal particles may be appropriately selected as required, and is not particularly limited as long as it can be dropped on a fluoride window and a compressed tablet can be produced. In some embodiments, the prescribed amount is, for example, 0.5 to 2. Mu.L, preferably 1. Mu.L.
Step (3)
In the invention, the step (3) is a step of covering the fluoride window with another fluoride window to be screwed into a tabletting, and observing the tabletting microsphere particle sample by an optical microscope and photographing. The reason for setting the step (3) is the same as that of the step (2), namely, by using a fluoride window sheet as a cover sheet when preparing a tablet, the problem of aggregation of microspherical particles can be effectively avoided, thereby ensuring that particles can be clearly distinguished after preparing an infrared transmission sample tablet, and a high-quality infrared spectrogram of specific particles can be clearly acquired.
In some embodiments of step (3), the optical microscopy observation is, for example, observation of the morphology, size, color of the sample of microspheroidal particles after tabletting using an optical microscope. Thereby, the selection of suitable microspheroidal particles for subsequent infrared transmission spectrogram determinations may be facilitated.
Step (4)
In the present invention, the step (4) is a step of observing the microspheroidal particles after tabletting by a fourier transform infrared microscope, taking a photograph, and measuring an infrared transmission spectrum of the specific microspheroidal particles. The reason for this step (4) is the following new findings of the present inventors:
for the microspheroidal particle samples prepared in the steps (1) - (3), if the reflection mode of the infrared spectrometer is used for testing, the signal in the reflection mode is reflected by the substrate after passing through the sample from the light source, and then the signal is transmitted back, so that the microspheroidal particles have higher thickness and are gathered on the substrate, the light path is longer, the energy loss is larger, and the microspheroidal particles are easy to move and difficult to fix, so that the test is unstable and the signal is poor, and even if a plurality of attempts are made, the effective spectrogram is difficult to obtain;
on the other hand, in the case of the sample of microspheroidal particles prepared in the above steps (1) to (3), if the test is conducted using the ATR mode of an infrared spectrometer, the effective spectrum is difficult to obtain because the ATR probe contacts the particle surface during the test, and the microspheroidal particles are likely to move during the test.
Accordingly, in view of the above new findings, the present inventors have improved the method of the prior art, and when detecting the main component of microspheroidal particles by infrared spectroscopy, the transmissive mode is used instead of other conventional modes such as the reflective mode, ATR mode, etc. Therefore, the obtained infrared transmission spectrogram can be ensured to have good matching degree with the standard spectrogram, thereby being beneficial to accurately identifying the main components of the microspherical particles.
In some embodiments of step (4), the fourier transform infrared microscope observation is an observation of the morphology, size of the sample of microspheroidal particles after tabletting using a fourier transform infrared microscope. Thereby, selection of suitable microspheroidal particles for infrared transmission spectrogram determination may be facilitated.
Step (5)
In the present invention, step (5) is a step of analyzing and comparing the obtained infrared transmission spectrum with a spectrum in a commercial database by using software, thereby identifying the main component of the sample. In the step (5), since the microspheroidal particle sample prepared by the method of the present invention is used, microspheroidal particles in the sample can be effectively distinguished and fixed, so that the infrared transmission spectrogram obtained by using the transmission mode has good matching degree with the standard spectrogram, and the detection and analysis comparison processes are rapid and efficient, and the main components of the microspheroidal particles can be accurately identified without multiple attempts.
In some embodiments of step (5), the software is infrared spectrum analysis software commonly used in the infrared spectroscopy arts, such as, for example, KIA software (WileySoftware), and the like.
In some embodiments of step (5), the search method used for the analytical comparison is a "first derivative Euclidean distance algorithm".
In some embodiments of step (5), the commercial database is a database commonly used in the infrared spectroscopy field, such as the Sadtler infrared spectroscopy database and the like.
Examples
The invention will be further illustrated with reference to specific examples. It is to be understood that these examples are illustrative of the present invention and are not intended to limit the scope of the present invention. Appropriate modifications and variations of the invention may be made by those skilled in the art, and are within the scope of the invention.
I. Preparation of a Transmission sample and determination of an Infrared Transmission Spectrometry
Two fillers based on agarose and methylol acrylic polymers were selected and their infrared transmission spectra were measured in transmission mode using the following method, respectively.
(a) The filler particles stored in the solution were observed with OM (optical microscope), and the filler particles were diluted 50-fold with ultrapure water: 10. Mu.L of the sample was measured using a pipette, 490. Mu.L of ultrapure water was added thereto, and the mixture was vortexed and homogenized for use.
(b) After mixing the samples, 1. Mu.L of diluted sample was measured and dropped on BaF using a pipette 2 On the (barium fluoride) window, baF 2 The (barium fluoride) window is arranged at the center of the groove at the lower layer of the micro-compression dish;
(c) Using another piece of BaF 2 The (barium fluoride) window sheet is covered on the window sheet with the filler sample, the window sheet is screwed to form a pressed sheet, the OM observation is carried out on the filler particle sample after the pressing sheet, and the photographing is carried out;
(d) The pellet after tabletting was observed by FTIR (Fourier transform Infrared microscope), a photograph was taken, and the filler pellet was subjected to infrared transmission spectrogram measurement with a scanning resolution of 4cm -1 The number of scans was 128. All infrared spectrograms need to be subjected to baseline and environmental impact correction before data comparison and analysis, including removal of moisture and CO 2 ；
(e) And (3) analyzing and comparing the obtained infrared spectrogram with spectrograms in a commercial database Sadtler by using KIA software, wherein the searching method is a first derivative Euclidean distance algorithm, so that the components of the sample are identified.
Comparison of other sample preparation and detection modes with the determination of IR spectra by the method of the present application
Two fillers based on agarose and methylol acrylic polymers were selected, and their infrared spectra were measured using reflection mode and ATR mode, respectively, and compared.
Reflection mode:
(i) Weighing 1 mu L of the filler sample diluted by 50 times and uniformly mixed by a pipetting gun, dripping the filler sample onto a silver-plated lens, and air-drying;
(ii) The selected filler particles were tested for infrared reflectance spectrum by selective reflection mode.
(iii) And (3) analyzing and comparing the obtained infrared spectrogram with spectrograms in a commercial database Sadtler by using KIA software, so as to identify the components of the sample.
ATR (attenuated total reflection) mode:
(i) Weighing 1 mu L of the filler sample diluted by 50 times and uniformly mixed by a pipetting gun, dripping the filler sample onto a silver-plated lens, and air-drying;
(ii) The ATR mode is selected to conduct infrared reflectance spectroscopy testing of selected filler particles.
(iii) And (3) analyzing and comparing the obtained infrared spectrogram with spectrograms in a commercial database Sadtler by using KIA software, so as to identify the components of the sample.
The main instrument and software information used in the examples is shown in table 1 below. The main consumables and material information used in the examples are shown in table 2 below.
TABLE 1 Main instruments/software used in the examples
| Numbering device | Instrument for measuring and controlling the intensity of light | Manufacturer/model |
| 1 | Optical microscope | Zeiss/Stemi 508 or similar instruments |
| 2 | Infrared microscope | bruk/Lumos or similar instruments |
| 3 | Knowtitall (software) | Wiley or other similar spectral processing software |
TABLE 2 consumable/material information used in the examples
Results
The infrared transmission samples were prepared by the above-described method for preparing transmission samples using filler particles of two different matrices, the schematic diagram of which is shown in fig. 1, and the OM (optical microscope) image of which is shown in fig. 2 and 3.
Samples were prepared on silver coated lenses according to the sample preparation method described above for the reflective and ATR (attenuated total reflection) mode samples with OM (optical microscopy) images as shown in fig. 4 and 5.
Comparing fig. 2 to 3 with fig. 4 to 5, it can be seen that when silver-plated lenses are used for sample preparation, filler particles are overlapped and aggregated together, and the particles cannot be distinguished, so that an infrared spectrogram of a specific particle cannot be acquired. Conversely, baF is used as described in the method of the present invention 2 When the (barium fluoride) window sheet is used for preparing samples, the filler particles can be well distinguished and fixed, so that the main components of the particles to be detected can be rapidly obtained by measuring the infrared transmission spectrogram of the filler particles through an infrared microscope.
In addition, the comparison results of the infrared spectrograms obtained in different infrared detection modes and the same standard spectrogram are shown in fig. 6 and 7. For filler particles with two different main components, the infrared spectrograms obtained in the reflection mode have larger differences from the standard spectrograms, and the main components of the filler particles cannot be identified or confirmed. The infrared spectra of the sample in the transmission mode and the sample in the ATR (attenuated total reflection) mode have high similarity with the standard spectrum, but comparing the infrared spectra of the sample in the transmission mode and the sample in the ATR (attenuated total reflection) mode shows that the sample using the ATR (attenuated total reflection) mode has the following two problems:
1) Because the filler particles are spherical, when the ATR (attenuated total reflection) detector gradually approaches and contacts the filler particles, the filler particles can shift, so that the ATR (attenuated total reflection) detector contacts a substrate (a gold-plated lens, a silver-plated lens or a gold-plated filtering film and the like), multiple attempts are needed, and the detection is time-consuming and low-efficiency;
2) When two or more different fillers are mixed or part of filler particles are abnormal (such as color change and the like), the filler particles cannot be distinguished, and the filler particles to be detected cannot be accurately positioned during testing.
The result shows that the infrared spectrogram obtained by preparing the transmission sample and using the transmission mode has good matching degree with the standard spectrogram, the detection process is quick and efficient, multiple attempts are not needed, and the main components of the spherical particles can be accurately detected. And the quality of the infrared spectrogram measured by the method is higher than the matching degree of the infrared spectrogram measured under other modes and the standard spectrogram, and the detection efficiency is high. Therefore, compared with the method that an undiluted sample is directly detected or an infrared spectrum is obtained by using a reflection mode or an ATR (attenuated total reflectance) mode of infrared spectrum, the method can obviously separate each filler microsphere, efficiently obtain the infrared spectrum of high-quality filler particles and further obtain main component related information of the filler through database comparison.
In summary, the above embodiments are only preferred embodiments of the present invention, and are not intended to limit the scope of the present invention, but any modifications, equivalents, improvements, etc. within the spirit and principles of the present invention should be included in the scope of the present invention.
Claims (10)
1. An analytical detection method of a main component of microspheroidal particles comprising the steps of:
(1) Diluting the microspherical particle sample by a specified multiple by using a diluent, and uniformly mixing for later use;
(2) Diluting and uniformly mixing the microspherical particle sample, measuring a specified amount of diluted microspherical particle sample, dripping the diluted microspherical particle sample on a fluoride window, and placing the fluoride window at the center of a groove at the lower layer of the micro-compression dish;
(3) Covering the fluoride window sheet with another fluoride window sheet, screwing the fluoride window sheet with the microsphere particle sample to prepare a tabletting, and observing the tabletting microsphere particle sample by using an optical microscope and photographing;
(4) Observing the pressed microspherical particles by using a Fourier transform infrared microscope, taking a picture, and measuring an infrared transmission spectrogram of the specific microspherical particles;
(5) And (3) analyzing and comparing the obtained infrared transmission spectrogram with spectrograms in a commercial database by using software, so as to identify the main components of the sample.
2. The method of claim 1, wherein the microspheroidal particles are chromatographic column packing particles.
3. The method of claim 1, wherein prior to said step (1), the morphology, size, color of said microspheroidal particles stored in solution are observed using an optical microscope.
4. The method according to claim 1, wherein the predetermined dilution ratio in the step (1) is 30 to 100 times.
5. The method of claim 1, wherein the mixing in step (1) is vortex mixing.
6. The method of claim 1, wherein the fluoride window in step (2) is barium fluoride BaF 2 Window pane, calcium fluoride CaF 2 Window or magnesium fluoride MgF 2 And a window sheet.
7. The method according to claim 1, wherein the diluted microspheroidal sample in step (2) is in the range of 0.5 to 2. Mu.L.
8. The method according to claim 1, wherein the optical microscopic observation in the step (3) is observation of morphology, size, and color of the sample of microspheroidal particles after tabletting by means of an optical microscope.
9. The method of claim 1, wherein the fourier transform infrared microscope observation in the step (4) is an observation of the morphology and size of the sample of microspheroidal particles after tabletting by using a fourier transform infrared microscope.
10. The method as claimed in claim 1The method wherein the software used in step (5) is KIA software, i.e., wileyThe search method used for analysis and comparison is a first derivative Euclidean distance algorithm, and the commercial database used is a Sadtler infrared spectrum database.
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