阅读说明：本技术 聚酰亚胺薄膜中9,9-双(4-氨基-3-氯苯基)芴的高效液相色谱检测方法 (High performance liquid chromatography detection method for 9, 9-bis (4-amino-3-chlorphenyl) fluorene in polyimide film ) 是由 张兰兰 钟琳 李支薇 杨文宇 严洪连 于 2020-12-21 设计创作，主要内容包括：本发明涉及色谱分析领域,特别是涉及一种薄膜中9,9-双(4-氨基-3-氯苯基)芴的高效液相色谱检测方法。本发明通过选用水和乙腈作为流动相,能高效地对经过萃取前处理得到的萃取液中的9,9-双(4-氨基-3-氯苯基)芴含量进行分析,进而确定薄膜中待测物含量。所得色谱峰峰型尖锐、对称性好,基线波动小,无漂移现象；选择合适的检测器波长,所得色谱峰信号强度合适,便于分析计算,减小误差,提高准确性。本发明的分析方法灵敏度和准确性高,检出限和定量限低,且色谱峰出峰保留时间早,分析效率高,能快速获得分析结果。(The invention relates to the field of chromatographic analysis, in particular to a high performance liquid chromatography detection method of 9, 9-bis (4-amino-3-chlorphenyl) fluorene in a film. According to the invention, water and acetonitrile are selected as mobile phases, so that the content of 9, 9-bis (4-amino-3-chlorphenyl) fluorene in the extraction liquid obtained by extraction pretreatment can be efficiently analyzed, and the content of a substance to be detected in the film can be further determined. The obtained chromatographic peak has sharp peak shape, good symmetry, small baseline fluctuation and no drift phenomenon; and the appropriate wavelength of the detector is selected, the obtained chromatographic peak signal intensity is appropriate, the analysis and calculation are convenient, the error is reduced, and the accuracy is improved. The analysis method has the advantages of high sensitivity and accuracy, low detection limit and quantification limit, early chromatographic peak-out retention time, high analysis efficiency and capability of quickly obtaining an analysis result.)

1. A method for detecting 9, 9-bis (4-amino-3-chlorphenyl) fluorene in a polyimide film is characterized by comprising the following steps:

extracting a film sample by using an organic solvent to obtain an extract, feeding the extract into a high performance liquid chromatograph, and performing isocratic elution, wherein a mobile phase is selected from water and acetonitrile, and the volume ratio of the water to the acetonitrile is (0.35-0.5) to (0.65-0.5).

2. The detection method according to claim 1, wherein an ultraviolet detector is selected for detection, and the detection wavelength is 180nm to 220 nm.

3. The detection method according to claim 1, wherein the ratio of the mass of the film sample to the volume of the organic solvent is 1g (8 mL-12 mL).

4. The detection method according to claim 1, wherein the organic solvent is at least one of dichloromethane, acetone, tetrahydrofuran, methanol, and acetonitrile.

5. The detection method according to claim 1, wherein the flow rate of the mobile phase is 1.2mL/min to 1.8 mL/min.

6. The detection method according to any one of claims 1 to 5, wherein the extraction operation is: and placing the film sample and the organic solvent into a container for ultrasonic treatment, wherein the ultrasonic power is 280W-520W, the ultrasonic frequency is 30 KHz-50 KHz, the ultrasonic temperature is more than or equal to 40 ℃, and the ultrasonic time is more than or equal to 40 minutes.

7. The detection method according to any one of claims 1 to 5, wherein the column of the high performance liquid chromatograph is C₁₈A chromatographic column.

8. The detection method according to claim 7, wherein the amount of the extract solution to be fed to the high performance liquid chromatograph is 5 to 20 μ L.

9. The detection method according to claim 7, wherein the column temperature of the chromatography column is 30 to 50 ℃.

10. The detection method according to any one of claims 1 to 5, wherein the film sample size is 2mm x 2mm or less.

11. The detection method according to any one of claims 1 to 5, wherein the extract is filtered before being fed to the high performance liquid chromatograph, and the pore size of a filter membrane is 0.22 μm or less.

Technical Field

The invention relates to the field of chromatographic analysis, in particular to a high performance liquid chromatography detection method of 9, 9-bis (4-amino-3-chlorphenyl) fluorene in a polyimide film.

Background

9, 9-bis (4-amino-3-chlorophenyl) fluorene is used as an important monomer raw material to synthesize polymer polymers such as polyimide, polyamide and polyurethane, wherein polyimide is the most common, has excellent thermal stability, chemical solvent resistance, good mechanical property and electrical property, and is mainly applied to the modern industrial fields such as aerospace, microelectronics, machinery and automobiles in the form of films and molded parts. However, the polyimide material has excellent industrial performance, but the residual 9, 9-bis (4-amino-3-chlorophenyl) fluorene can bring harm to the health of people, and researches prove that the compound has strong irritant effect on skin and eyes and persistent toxicity on aquatic organisms, so that a proper analysis method is necessary for determining the content of the 9, 9-bis (4-amino-3-chlorophenyl) fluorene in the product. However, in the current research on 9, 9-bis (4-amino-3-chlorophenyl) fluorene, synthesis and application of the 9, 9-bis (4-amino-3-chlorophenyl) fluorene and related derivatives are mostly concerned, and no reports are found on an analysis method for the content of the residual 9, 9-bis (4-amino-3-chlorophenyl) fluorene in the material.

Disclosure of Invention

Based on the above, it is necessary to provide a rapid, accurate and highly sensitive detection method for detecting the content of 9, 9-bis (4-amino-3-chlorophenyl) fluorene in a polyimide film.

The invention provides a method for detecting 9, 9-bis (4-amino-3-chlorphenyl) fluorene in a polyimide film, which comprises the following steps:

extracting a film sample by using an organic solvent to obtain an extract, feeding the extract into a high performance liquid chromatograph, and performing isocratic elution, wherein a mobile phase is selected from water and acetonitrile, and the volume ratio of the water to the acetonitrile is (0.35-0.5) to (0.65-0.5).

In one embodiment, an ultraviolet detector is selected for detection, and the detection wavelength is 180 nm-220 nm.

In one embodiment, the ratio of the mass of the film sample to the volume of the organic solvent is 1g (8 mL-12 mL).

In one embodiment, the organic solvent is at least one of dichloromethane, acetone, tetrahydrofuran, methanol, and acetonitrile.

In one embodiment, the flow rate of the mobile phase is 1.2mL/min to 1.8 mL/min.

In one embodiment, the extraction operation is: and placing the film sample and the organic solvent into a container for ultrasonic treatment, wherein the ultrasonic power is 280W-520W, the ultrasonic frequency is 30 KHz-50 KHz, the ultrasonic temperature is more than or equal to 40 ℃, and the ultrasonic time is more than or equal to 40 minutes.

In one embodiment, the chromatographic column of the high performance liquid chromatograph is C₁₈A chromatographic column.

In one embodiment, the sample amount of the extract liquid to be fed to the high performance liquid chromatograph is 5-20 μ L.

In one embodiment, the temperature of the chromatographic column is 30 ℃ to 50 ℃.

In one embodiment, the film sample size is 2mm x 2mm or less.

In one embodiment, the extract is filtered before being fed into the high performance liquid chromatograph, and the pore diameter of a filter membrane is less than or equal to 0.22 μm.

According to the invention, water and acetonitrile are selected as mobile phases, and the volume ratio of the water to the acetonitrile is set to (0.35-0.5) to (0.65-0.5), so that the content of 9, 9-bis (4-amino-3-chlorphenyl) fluorene in the extraction liquid obtained by extraction pretreatment can be efficiently analyzed, and the content of 9, 9-bis (4-amino-3-chlorphenyl) fluorene in the polyimide film can be further determined. The obtained chromatographic peak has sharp peak shape, good symmetry, small baseline fluctuation and no drift phenomenon; and a proper detector wavelength is selected, so that the obtained chromatographic peak signal intensity is proper, the analysis and calculation are convenient, and the accuracy is improved. The analysis method has the advantages of high sensitivity and accuracy, low detection limit and quantification limit, early chromatographic peak-out retention time, high analysis efficiency and capability of quickly obtaining an analysis result.

Drawings

FIG. 1 is a high performance liquid chromatogram of example 1;

FIG. 2 is a high performance liquid chromatogram of comparative example 1.

Detailed Description

To facilitate an understanding of the invention, the invention will now be described more fully with reference to the accompanying drawings. Preferred embodiments of the present invention are shown in the drawings. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.

Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one such feature. In the description of the invention, "a plurality" means at least two, e.g., two, three, etc., unless explicitly specified otherwise. In the description of the present invention, "a plurality" means at least one, e.g., one, two, etc., unless specifically limited otherwise.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

The invention provides a method for detecting 9, 9-bis (4-amino-3-chlorphenyl) fluorene in a polyimide film, which comprises the following steps:

extracting a film sample by using an organic solvent to obtain an extract, feeding the extract into a high performance liquid chromatograph, and carrying out isocratic elution, wherein water and acetonitrile are selected as mobile phases, the volume ratio of the water to the acetonitrile is (0.35-0.5) - (0.65-0.5), preferably, the volume ratio of the water to the acetonitrile is 0.4:0.6, and proper mobile phase proportion and gradient setting are directly related to the quality of a chromatographic peak shape, the length of retention time and whether the chromatographic peak shape can be well separated from impurities.

Methanol and acetonitrile are respectively adopted as organic phases, water is adopted as a water phase, and the organic phases and the water phases are combined pairwise to optimize conditions. The result shows that when methanol is selected as the mobile phase organic phase, the peak emergence time of the 9, 9-bis (4-amino-3-chlorphenyl) fluorene is late, and the peak width is wide; when acetonitrile is selected as a mobile phase organic phase, the 9, 9-bis (4-amino-3-chlorphenyl) fluorene has symmetrical peak shape, narrow peak width, high response, early peak emergence time and no impurity peak interference near the retention time, and cannot influence analysis.

High performance liquid chromatography usually uses gradient elution to achieve good separation of each component in a complex sample according to a suitable capacity factor (k), so that the mobile phase water (a) and acetonitrile (B) used in the initial experiment are analyzed by gradient elution, and one of the gradient elution ratios is tried: within 0-2 min, the proportion of acetonitrile is 50%; within 2-5 min, gradually increasing the proportion of acetonitrile from 50% to 80%, and keeping for 5 min; and within 10-12 min, gradually reducing the proportion of acetonitrile from 80% to 50%, and keeping for 3 min. Under the elution condition, the baseline of the liquid chromatogram fluctuates, and the whole liquid chromatogram drifts upwards, thereby influencing the peak shape of the chromatographic peak. In addition, the elution ratio is adjusted, gradient elution is performed at other ratios, but the base line still drifts upwards, so that the substance to be detected is determined to be analyzed by adopting an isocratic elution mode. Under the optimized chromatographic conditions, namely the volume ratio of water to acetonitrile is (0.35-0.5) to (0.65-0.5), the 9, 9-bis (4-amino-3-chlorphenyl) fluorene has early appearance time, the chromatographic peak shape is sharp and symmetrical, and no impurity peak interference exists near the retention time.

In one specific example, the ultraviolet detector is selected to detect light with a wavelength of 180nm to 220nm, preferably 200 nm. The proper wavelength of the detector can ensure that the intensity of the peak signal of the chromatogram is proper, the analysis and calculation are convenient, and the error is reduced, so that the accuracy of the analysis and detection method is improved.

In one specific example, the ratio of the mass of the film sample to the volume of the organic solvent is 1g (8 mL-12 mL), and preferably the ratio of the mass of the film sample to the volume of the organic solvent is 1g:10 mL. The proper amount of the extractant can improve the extraction efficiency without causing the concentration of the substance to be detected to be too low and requiring an additional concentration step.

In a specific example, the organic solvent is at least one of dichloromethane, acetone, tetrahydrofuran, methanol, and acetonitrile. Preferably, acetonitrile is selected as the extracting agent, the average extraction efficiency is greater than or equal to 85%, and the test requirement can be better met.

In one specific example, the flow rate of the mobile phase is 1.2mL/min to 1.8mL/min, preferably the flow rate of the mobile phase is 1.4mL/min to 1.6mL/min, and more preferably the flow rate of the mobile phase is 1.5 mL/min. The appropriate flow velocity of the mobile phase can balance the column efficiency and the analysis time, so that the analysis method has higher column efficiency, and meanwhile, the analysis time is not too long, and the analysis efficiency is not influenced.

In one particular example, the extraction operation is: placing the film sample and an organic solvent into a container for ultrasonic treatment, wherein the ultrasonic power is 280W-520W, the ultrasonic frequency is 30 KHz-50 KHz, the ultrasonic temperature is more than or equal to 40 ℃, the ultrasonic time is more than or equal to 40 minutes, and the extraction efficiency is 75 percent; preferably, the ultrasonic temperature is greater than or equal to 50 ℃, the ultrasonic time is greater than or equal to 50 minutes, and the extraction efficiency is 80 percent; more preferably, the ultrasonic temperature is more than or equal to 60 ℃, the ultrasonic time is more than or equal to 70 minutes, and under the extraction condition, the extraction efficiency of more than 85 percent can be achieved.

In one specific example, the column of the high performance liquid chromatograph is C₁₈A chromatographic column; preferably, the column specifications are: the column length is 250mm, the diameter is 4.6mm, and the filler particle size is 5 μm.

In a specific example, the extraction liquid is injected into the high performance liquid chromatograph in an amount of 5 μ L to 20 μ L, preferably in an amount of 8 μ L to 12 μ L, and more preferably in an amount of 10 μ L. The appropriate sample injection amount can enable the intensity of chromatographic peak signals to be moderate, the signals are not too strong and can not respond to impurities, and the signals are not too weak and baseline drift can not affect the peak type, so that the error is too large, and the accuracy of an analysis result is reduced.

In a specific example, the column temperature of the chromatography column is 30 ℃ to 50 ℃, preferably 35 ℃ to 45 ℃, and more preferably 40 ℃. The proper column temperature can realize better separation effect, and on the premise of ensuring the separation effect, the retention time of the substance to be detected is shorter, and the separation efficiency is improved.

In one specific example, the size of the film sample is 2mm × 2mm or less, and the extraction efficiency can be made higher by maintaining the size of the film sample below 2mm × 2 mm.

In a specific example, before the extraction liquid is fed into the high performance liquid chromatograph, the extraction liquid is filtered, the aperture of the filter membrane is less than or equal to 0.22 μm, and the filtering before the feeding can prevent large particles in the extraction liquid from causing negative influence on the analysis process or the high performance liquid chromatograph.

According to the invention, water and acetonitrile are selected as mobile phases, and the volume ratio of the water to the acetonitrile is set to (0.35-0.5) to (0.65-0.5), so that the content of 9, 9-bis (4-amino-3-chlorphenyl) fluorene in the extraction liquid obtained by extraction pretreatment can be efficiently analyzed, and the content of 9, 9-bis (4-amino-3-chlorphenyl) fluorene in the film can be further determined. The obtained chromatographic peak has sharp peak shape, good symmetry, small baseline fluctuation and no drift phenomenon; and the proper wavelength of the detector is selected, so that the intensity of the obtained chromatographic peak signal is proper, the analysis and calculation are convenient, and the analysis accuracy is improved. The analysis method provided by the invention has the advantages of high sensitivity, low detection limit and quantification limit, early chromatographic peak-appearance retention time, high analysis efficiency and capability of rapidly obtaining an analysis result.

The present invention will be described in further detail with reference to specific examples and comparative examples. It is understood that the following examples are more specific in terms of apparatus and materials, and in other embodiments, are not limited thereto, such as, but not limited to, Agilent ZORBAX SB-Aq columns available from Agilent technologies, Inc., USA.

9, 9-bis (4-amino-3-chlorophenyl) standard: 9, 9-bis (4-amino-3-chlorophenyl) with a purity of not less than 98.0%, Tokyo Kasei Kogyo;

other reagents: acetonitrile, methanol, acetone, tetrahydrofuran and dichloromethane, all HPLC grade, Sigma company, usa;

high performance liquid chromatograph: LC-20A high performance liquid chromatography with UV detector, Shimadzu corporation, Japan;

a chromatographic column: agilent ZORBAX SB-Aq, Agilent technologies, Inc., USA;

ultrasonic cleaner: 2300HT, shanghai' an spectral science instruments ltd;

precision electronic analytical balance: XA205DU, sensory 0.1mg, Metler Switzerland;

a vortex mixer: XW-80A, Shanghai Jingke industries, Ltd;

an ultra-pure water machine: Milli-Q, Millipore USA.

Example 1

1. Taking a film sample, shearing the film sample until the size of the sample is less than or equal to 2mm multiplied by 2mm, accurately weighing 1g of the sample (accurate to 0.1mg) and placing the sample into a sample bottle, adding 10mL of acetonitrile into the sample bottle, screwing a bottle cap, placing the sample bottle into an ultrasonic cleaning instrument, and performing ultrasonic treatment at 60 ℃ for 70 min. And filtering 1mL of cooled extract liquor through a 0.22-micron filter membrane to obtain a sample to be detected.

2. Setting the column temperature of a high performance liquid chromatograph at 40 ℃, the detection wavelength at 200nm, selecting water and acetonitrile as mobile phases, setting the volume ratio of the water to the acetonitrile at 0.4:0.6, setting the flow rate of the mobile phases at 1.5mL/min, then taking 10 mu L of a sample to be detected, injecting the sample, and carrying out isocratic elution.

And after the analysis is finished, a liquid chromatographic peak is obtained, the retention time of the 9, 9-bis (4-amino-3-chlorphenyl) fluorene is 6.68min, the peak shape is sharp, symmetrical and not trailing, no interference peak is generated near the main peak, accurate quantitative analysis can be carried out, and the concentration of the 9, 9-bis (4-amino-3-chlorphenyl) fluorene in the sample is 10 mg/L.

Comparative example 1

1. Taking a film sample, shearing the film sample until the size of the sample is less than or equal to 2mm multiplied by 2mm, accurately weighing 1g of the sample (accurate to 0.1mg) and placing the sample into a sample bottle, adding 10mL of acetonitrile into the sample bottle, screwing a bottle cap, placing the sample bottle into an ultrasonic cleaning instrument, and performing ultrasonic treatment at 60 ℃ for 70 min. And filtering 1mL of cooled extract liquor through a 0.22-micron filter membrane to obtain a sample to be detected.

2. Setting the column temperature of a high performance liquid chromatograph to be 40 ℃, the detection wavelength to be 200nm, selecting water and acetonitrile as mobile phases, wherein the flow rate of the mobile phases is 1.5mL/min, then taking 10 mu L of a sample to be detected, carrying out gradient elution, and setting the gradient as follows: within 0-2 min, the proportion of acetonitrile is 50%; gradually increasing from 50% to 80% within 2-5 min, and keeping for 5 min; within 10-12 min, the proportion is gradually reduced from 80% to 50%, and the temperature is kept for 3 min.

And (4) obtaining a liquid chromatogram peak after the analysis is finished, wherein the baseline of the liquid chromatogram fluctuates and integrally drifts upwards under the elution condition, so that accurate quantitative analysis cannot be carried out.

Comparative example 2

1. Taking a film sample, shearing the film sample until the size of the sample is less than or equal to 2mm multiplied by 2mm, accurately weighing 1g of the sample (accurate to 0.1mg) and placing the sample into a sample bottle, adding 10mL of acetonitrile into the sample bottle, screwing a bottle cap, placing the sample bottle into an ultrasonic cleaning instrument, and performing ultrasonic treatment at 60 ℃ for 70 min. And filtering 1mL of cooled extract liquor through a 0.22-micron filter membrane to obtain a sample to be detected.

2. Setting the column temperature of a high performance liquid chromatograph at 40 ℃, the detection wavelength at 200nm, selecting water and acetonitrile as mobile phases, setting the volume ratio of the water to the acetonitrile at 0.3:0.7, setting the flow rate of the mobile phases at 1.5mL/min, then taking 10 mu L of a sample to be detected, injecting the sample, and carrying out isocratic elution.

And after the analysis is finished, obtaining a liquid chromatographic peak, wherein under the condition, the retention time of the 9, 9-bis (4-amino-3-chlorphenyl) fluorene is only 3.92min, and the peak emergence time of an interference peak in a film sample is earlier, so that the premature retention time of the object to be detected enables the main peak and the interference peak to be easily overlapped in the analysis process, and accurate quantitative analysis cannot be carried out.

Comparative example 3

Taking a film sample, shearing the film sample until the size of the sample is less than or equal to 2mm multiplied by 2mm, accurately weighing 1g of the sample (accurate to 0.1mg) and putting the sample into a sample bottle, adding 10mL of acetonitrile into the sample bottle, screwing a bottle cap, putting the sample bottle into an ultrasonic cleaning instrument, performing ultrasonic treatment at 30 ℃ for 70min, wherein the extraction efficiency is only 54%, and the test requirement cannot be met.

Comparative example 4

Taking a film sample, shearing the film sample until the size of the sample is less than or equal to 2mm multiplied by 2mm, accurately weighing 1g of the sample (accurate to 0.1mg) and putting the sample into a sample bottle, adding 10mL of acetonitrile into the sample bottle, screwing a bottle cap, putting the sample bottle into an ultrasonic cleaning instrument, performing ultrasonic treatment at 60 ℃ for 30min, wherein the extraction efficiency is only 65%, and the test requirement cannot be met.

Determination of linear equation, correlation coefficient, detection limit and quantification limit:

1. preparation of the solution

Standard stock solutions: accurately weighing 10mg (accurate to 0.1mg) of standard substance, dissolving with acetonitrile, and diluting to a volume of 10mL volumetric flask to obtain 1000mg/L stock solution of standard substance, and storing in a refrigerator at 4 deg.C in dark place.

Standard working solution: the standard stock solution was diluted with acetonitrile stepwise to obtain 0.1mg/L, 0.2mg/L, 0.5mg/L, 1.0mg/L, 2.0mg/L series of standard working solutions.

2. Drawing of standard working curve

Injecting a standard working solution, analyzing according to the analysis method described in the embodiment 1, calculating a peak area after the analysis is finished, and then drawing a standard working curve by taking the mass concentration X (mg/L) as a horizontal coordinate and the peak area Y as a vertical coordinate, wherein the obtained linear equation is as follows: y ═ 7.9 × 10⁴X +153.7 with a correlation coefficient of 0.9999; on the basis, the detection limit of the standard substance is determined to be 0.011mg/L by 3 times of signal-to-noise ratio, and the quantitative limit of the standard substance is determined to be 0.037mg/L by 10 times of signal-to-noise ratio.

Therefore, the analysis method is in the linear range of 0.1 mg/L-2.0 mg/L, the 9, 9-bis (4-amino-3-chlorphenyl) fluorene shows good linear relation, the correlation coefficient is 0.9999, the detection limit and the quantification limit are low, and the sensitivity is high.

Determination of recovery and precision:

the recovery rate research experiment is carried out by a blank matrix standard adding experiment, a sample without the to-be-detected 9, 9-bis (4-amino-3-chlorphenyl) fluorene is selected as a blank matrix, and 3 different standard adding concentration levels (0.1mg/L, 0.5mg/L and 2.0mg/L) are set. Each spiked level was tested in parallel 6 times (n-6) for precision. Under 3 standard adding levels, the standard adding recovery rate of 9, 9-bis (4-amino-3-chlorphenyl) fluorene is 98.2% -100.4%, and the relative standard deviation of the recovery rate is not more than 2.8%, which shows that the test method has high accuracy and can meet the test requirements.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.