CN115032318A - Liquid chromatography analysis method for detecting content of 3- (N-nitrosomethylamino) propionitrile - Google Patents
Liquid chromatography analysis method for detecting content of 3- (N-nitrosomethylamino) propionitrile Download PDFInfo
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- HZDLDFBKYBGNHG-UHFFFAOYSA-N 3-(N-Nitrosomethylamino)propionitrile Chemical compound O=NN(C)CCC#N HZDLDFBKYBGNHG-UHFFFAOYSA-N 0.000 title claims abstract description 43
- 238000004811 liquid chromatography Methods 0.000 title claims abstract description 23
- 238000004458 analytical method Methods 0.000 title claims abstract description 22
- 238000001514 detection method Methods 0.000 claims abstract description 32
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- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 6
- 235000019253 formic acid Nutrition 0.000 claims description 6
- 239000000741 silica gel Substances 0.000 claims description 6
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- XKLJHFLUAHKGGU-UHFFFAOYSA-N nitrous amide Chemical compound ON=N XKLJHFLUAHKGGU-UHFFFAOYSA-N 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/86—Signal analysis
- G01N30/8624—Detection of slopes or peaks; baseline correction
- G01N30/8631—Peaks
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- Life Sciences & Earth Sciences (AREA)
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- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
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Abstract
The present disclosure relates to a liquid chromatography method for detecting the content of 3- (N-nitrosomethylamino) propionitrile, which comprises the following steps: (1) preparing a standard solution and determining a standard curve for 3- (N-nitrosomethylamino) propionitrile, in which Y is the sum of peak areas corresponding to 5.50-6.00min and 6.30-6.80 min; (2) and detecting the sample to be detected by using a liquid chromatography analysis method, wherein in the obtained chromatogram, the peak area corresponding to the target is the sum of the peak areas corresponding to 5.50-6.00min and 6.30-6.80 min. The analysis method provided by the disclosure creatively incorporates isomers into the detection of the content of the 3- (N-nitrosomethylamino) propionitrile, and can avoid the phenomenon of false qualification to a great extent.
Description
Technical Field
The disclosure relates to the technical field of compound content detection, in particular to a liquid chromatography method for detecting the content of 3- (N-nitrosomethylamino) propionitrile, and especially relates to a liquid chromatography method for detecting the content of 3- (N-nitrosomethylamino) propionitrile in betel nuts.
Background
Nitrosamine impurities belong to the "cohort of interest" substances mentioned in the guidance of ICH M7(R1) ("assessing and controlling DNA reactive (mutagenic) impurities in drugs to limit potential carcinogenic risks"). According to the carcinogen list published by the world health organization, 3- (N-nitrosomethylamino) propionitrile is a class 2B carcinogen.
The structural formula of 3- (N-nitrosomethylamino) propionitrile is as follows:
areca catechu contains various nutrient elements and beneficial substances required by human body, and has the efficacies of killing parasites, breaking food retention, reducing qi, activating stagnancy, promoting diuresis and resolving dampness, so that the areca catechu is used for treating parasitic infections such as tapeworm, hookworm, roundworm, fasciola gigantica and fasciolopsis and the like, and the Simotang decoction containing the areca catechu is mainly used for treating seven emotions, acute asthma, stuffiness and no eating. Betel nut has high economic value and can be deeply processed into various products, but the ingredient contains a certain amount of 3- (N-nitrosomethylamino) propionitrile, and the 3- (N-nitrosomethylamino) propionitrile is classified as a 2B carcinogen, so the content of the 3- (N-nitrosomethylamino) propionitrile in the product needs to be limited, and the detection of the 3- (N-nitrosomethylamino) propionitrile is less and the phenomenon of false qualification is easy to occur at present.
Therefore, it is desirable to provide a method capable of accurately detecting the content of 3- (N-nitrosomethylamino) propionitrile.
Disclosure of Invention
In order to solve the technical problem, the present disclosure provides a liquid chromatography method for detecting the content of 3- (N-nitrosomethylamino) propionitrile.
In a first aspect, the present disclosure provides a liquid chromatography method for detecting the content of 3- (N-nitrosomethylamino) propionitrile, the liquid chromatography method comprising the steps of:
(1) preparing a standard solution and determining a standard curve for 3- (N-nitrosomethylamino) propionitrile, in which Y is the sum of peak areas corresponding to 5.50-6.00min and 6.30-6.80 min;
(2) and detecting the sample to be detected by using a liquid chromatographic analysis method, wherein in the obtained chromatogram, the peak area corresponding to the target is the sum of the peak areas corresponding to 5.50-6.00min and 6.30-6.80 min.
In the chromatogram obtained by the liquid chromatography detection of the standard solution, in view of normal fluctuation within an allowable range due to different liquid chromatographs, different detection times, different detection personnel and the like, the peak appearance time of the chromatographic peak of the 3- (N-nitrosomethylamino) propionitrile is limited to 5.50-6.00min, and the peak appearance time of the isomer thereof is limited to 6.30-6.80 min.
In the existing detection of 3- (N-nitrosomethylamino) propionitrile, only the content of a target object is generally detected, but the disclosure finds that the 3- (N-nitrosomethylamino) propionitrile and the isomer thereof can be mutually converted under certain conditions, and the conversion conditions are extremely simple, so that in the detection of the content of the 3- (N-nitrosomethylamino) propionitrile, the concentration of the 3- (N-nitrosomethylamino) propionitrile isomer is creatively incorporated into the detection method provided by the disclosure, and further, the condition of inaccurate detection caused by the mutual conversion of the 3- (N-nitrosomethylamino) propionitrile and the isomer thereof after the detection is finished is avoided.
In the present disclosure, the liquid chromatography detection parameters include:
the chromatographic column is Thermo Hypersil GOLD TM The mobile phase is an aqueous solution containing acetonitrile and formic acid, the flow rate is 0.8-1.2mL/min, the detection wavelength is 230nm, and the column temperature is 35-45 ℃.
It has been found by the present disclosure that although the maximum absorption wavelength of 3- (N-nitrosomethylamino) propionitrile is 226.6nm and the maximum absorption wavelength of its isomer is 231.3nm, the present disclosure prefers a detection wavelength of 230nm, in which case the ratio of 3- (N-nitrosomethylamino) propionitrile to its isomer is about 80:20, which can be mutually verified with the nuclear magnetic detection results, and thus the detection of the sample is more accurate.
As a preferred embodiment of the present disclosure, the volume ratio of acetonitrile, formic acid and water in the mobile phase is 500:1: 9500.
As a preferred technical scheme of the present disclosure, the standard solution contains 3- (N-nitrosomethylamino) propionitrile standard substance with 6 grades of concentration.
The 6 grades of concentration described in this disclosure are 4.5. mu.g/mL, 9.0. mu.g/mL, 22.0. mu.g/mL, 44.0. mu.g/mL, 110.0. mu.g/mL, 440.0. mu.g/mL, respectively, while pure solvents can also be included in the standard solution.
As a preferred technical solution of the present disclosure, the pretreatment method of the sample to be tested includes: the pretreatment method of the sample to be detected comprises the following steps: and extracting, concentrating, eluting by a silica gel column, concentrating again, separating by a chromatographic column, and drying to obtain a sample for detection and analysis.
As a preferred embodiment of the present disclosure, the present disclosure now provides the following pretreatment methods:
when the detected sample is a solid sample, crushing the sample, then continuously extracting for 6 hours by using ethyl acetate, and after the extraction is finished, using MgSO (MgSO) for the ethyl acetate 4 Drying, concentrating by rotary evaporation to about 2 mL; eluting the concentrated sample with 100mL ethyl acetate on a 2-x15-cm silica gel column (40-140 mesh), evaporating and concentrating the eluent to about 1mL, and separating by thin layer chromatography (silica gel, 2 mm; chromatography liquid ether: methanol ═ 9: 1); the band corresponding to 3- (N-nitrosomethylamino) propionitrile was extracted with dichloromethane, concentrated to about 1mL, and finally dried with a kudena-danish device, and each sample was dissolved in 50. mu.L of ethyl acetate and analyzed with GC-TEA.
When the detection sample is a liquid sample, collecting the liquid sample, storing the liquid sample in a mixed solution of 50mL of acetone and water (50:50) at the temperature of 0-5 ℃, and completing analysis within 48 h; the preserved samples were continuously extracted with ethyl acetate for 6h, followed by pretreatment of reference solid samples.
The analysis method provided by the present disclosure can detect a very small amount of a target, and as a preferred embodiment of the present disclosure, in the analysis method, the detection limit concentration corresponding to 3 times of noise is 0.37 μ g/mL.
Compared with the prior art, the technical scheme provided by the embodiment of the disclosure has the following advantages:
the analysis method provided by the disclosure creatively incorporates the isomer into the detection of the content of the 3- (N-nitrosomethylamino) propionitrile, and can greatly avoid the phenomenon of false qualification.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the present disclosure and together with the description, serve to explain the principles of the disclosure.
In order to more clearly illustrate the embodiments or technical solutions in the prior art of the present disclosure, the drawings used in the description of the embodiments or prior art will be briefly described below, and it is obvious for those skilled in the art that other drawings can be obtained according to the drawings without inventive exercise.
FIG. 1 is a chromatogram obtained by assaying a standard solution (1.1. mu.g/mL) according to an embodiment of the present disclosure.
Detailed Description
In order that the above objects, features and advantages of the present disclosure may be more clearly understood, aspects of the present disclosure will be further described below. It should be noted that the embodiments and features of the embodiments of the present disclosure may be combined with each other without conflict.
In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present disclosure, but the present disclosure may be practiced in other ways than those described herein; it is to be understood that the embodiments disclosed in the specification are only a few embodiments of the present disclosure, and not all embodiments.
Example 1
This example provides a liquid chromatography method for detecting the content of 3- (N-nitrosomethylamino) propionitrile, as follows:
(1) dissolving a 3- (N-nitrosomethylamino) propionitrile standard substance by using an acetonitrile solvent to prepare standard solutions with the concentrations of 0 mu g/mL, 4.5 mu g/mL, 9.0 mu g/mL, 22.0 mu g/mL, 44.0 mu g/mL, 110.0 mu g/mL and 440.0 mu g/mL respectively;
(2) measuring the standard solution by using a liquid chromatography analysis method, repeatedly measuring each concentration for 2 times, performing linear regression on the concentration X by using a peak area Y, and drawing a standard curve;
the liquid chromatograph used in this example was: shimadzu LC-20A liquid chromatograph with Thermo Hypersil GOLD as chromatographic column TM The mobile phase is an aqueous solution containing acetonitrile and formic acid (the volume ratio of the acetonitrile to the formic acid to the water is 500:1:9500), the flow rate is 1mL/min, the detection wavelength is 230nm, and the column temperature is 35-45 ℃.
FIG. 1 is a chromatogram obtained by measuring a standard solution with a concentration of 1.1. mu.g/mL in this example, wherein the chromatographic peak at 5.847min is 3- (N-nitrosomethylamino) propionitrile, and the chromatographic peak at 6.547min is an isomer thereof;
when a standard curve is determined, Y is the sum of peak areas of a chromatographic peak at 5.50-6.00min and a chromatographic peak at 6.30-6.80min, X is the concentration of the standard solution, the obtained standard curve is drawn to be Y-18726.72X +4764.56, and the correlation coefficient r-1;
(3) pretreating a sample to be detected:
pulverizing the sample, extracting with ethyl acetate for 6 hr, and collecting the ethyl acetate with MgSO 4 Drying, concentrating by rotary evaporation to about 2 mL; eluting the concentrated sample with 100mL ethyl acetate on a 2-x15-cm silica gel column (40-140 mesh), evaporating and concentrating the eluent to about 1mL, and separating by thin layer chromatography (silica gel, 2 mm; chromatography liquid ether: methanol ═ 9: 1); the band corresponding to 3- (N-nitrosomethylamino) propionitrile was extracted with dichloromethane, concentrated to about 1mL, and finally dried with a kudena-danish device, and each sample was dissolved in 50. mu.L of ethyl acetate for analysis;
(4) detecting the sample to be detected by using a liquid chromatography analysis method, wherein the peak area corresponding to the target is the sum of the peak area at 5.50-6.00min and the peak area at 6.30-6.80min, and substituting the peak area into a standard curve to obtain the concentration of the target (3- (N-nitrosomethylamino) propionitrile) in the sample to be detected.
Comparative example 1
This comparative example provides a liquid chromatography method for detecting the content of 3- (N-nitrosomethylamino) propionitrile.
The difference from example 1 is that in this comparative example, the peak area corresponding to Y is a peak area of 5.50 to 6.00min, while the peak area corresponding to the object is a peak area at 5.50 to 6.00 min.
Performance analysis 1: accuracy of analytical methods
3- (N-nitrosomethylamino) propionitrile was added to the blank substrate to control the concentration of 3- (N-nitrosomethylamino) propionitrile at 446.8. mu.g/mL, 450.8. mu.g/mL, 436.0. mu.g/mL, and the contents of the target substances were measured by the methods provided in example 1 and comparative example 1, respectively, and the results are shown in Table 1:
TABLE 1
As can be seen from table 1, the method provided by the present disclosure can accurately detect the concentration of the target substance, and if the target substance isomer is not included in the detection method, there is a possibility that the content of 3- (N-nitrosomethylamino) propionitrile in the sample to be detected is "false-qualified" due to the interconversion between the target substance and the isomer.
Performance test 2
(1) Detection limit
The signal-to-noise ratio measurements of 3- (N-nitrosomethylamino) propionitrile isomers at a concentration of 4.5. mu.g/mL were 37.12 and 34.14, respectively, and the concentration corresponding to a signal-to-noise ratio of 3 was calculated based on the standard curve obtained in example 1, and from the above performance tests, the detection limit concentration corresponding to 3 times the noise was 0.37. mu.g/mL when the detection limit was determined by the noise method.
(2) Precision degree
The sample is prepared into a solution with the mass concentration of 1.108 mu g/mL, 5 needles are continuously injected by adopting the method, and the RSD is 1.64 percent by peak area calculation.
Performance test 3
Preparing 3- (N-nitrosomethylamino) propionitrile standard into sample capable of performing nuclear magnetic detection, standing at 30 deg.C for 4 hr, detecting, and respectively determining 1 H、 13 C、 1 H- 1 H COSY、 1 H- 13 C HSQC、 1 H- 13 C HMBC、 1 H- 15 N HMBC spectrum, and as a result, the following results are found:
the obtained nuclear magnetic spectrogram shows that the sample comprises two substances, namely 3- (N-nitrosomethylamino) propionitrile and an isomer thereof, and shows that even if the sample to be detected is a 3- (N-nitrosomethylamino) propionitrile standard product, partial components are still converted into the isomer during detection, and the conversion condition is extremely simple. From this, it is found that, when the content of the target substance is detected only and the isomer content is ignored to determine that the product is acceptable, if the isomer is converted into the target substance, a phenomenon of "false acceptance" is very likely to occur, resulting in certain adverse effects.
Therefore, the analysis method provided by the disclosure creatively incorporates the isomer into the detection of the content of the 3- (N-nitrosomethylamino) propionitrile, and can avoid the phenomenon of false qualification to a great extent.
It is noted that, in this document, relational terms such as "first" and "second," and the like, may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising an … …" does not exclude the presence of other identical elements in a process, method, article, or apparatus that comprises the element.
The foregoing are merely exemplary embodiments of the present disclosure, which enable those skilled in the art to understand or practice the present disclosure. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the disclosure. Thus, the present disclosure is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (6)
1. A liquid chromatography method for detecting the content of 3- (N-nitrosomethylamino) propionitrile, which comprises the following steps:
(1) preparing a standard solution and determining a standard curve for 3- (N-nitrosomethylamino) propionitrile, in which Y is the sum of peak areas corresponding to 5.50-6.00min and 6.30-6.80 min;
(2) and detecting the sample to be detected by using a liquid chromatography analysis method, wherein in the obtained chromatogram, the peak area corresponding to the target is the sum of the peak areas corresponding to 5.50-6.00min and 6.30-6.80 min.
2. The liquid chromatography analysis method of claim 1, wherein the liquid chromatography detection parameters comprise:
the chromatographic column is Thermo Hypersil GOLD TM The mobile phase is an aqueous solution containing acetonitrile and formic acid, the flow rate is 0.8-1.2mL/min, the detection wavelength is 230nm, and the column temperature is 35-45 ℃.
3. The liquid chromatography method of claim 2, wherein the volume ratio of acetonitrile, formic acid and water in the mobile phase is 500:1: 9500.
4. The liquid chromatography analysis method according to any one of claims 1 to 3, wherein the standard solution contains 3- (N-nitrosomethylamino) propionitrile standards at 6-order concentrations.
5. The liquid chromatography analysis method according to any one of claims 1 to 4, wherein the pretreatment method of the sample to be tested comprises: and extracting, concentrating, eluting by a silica gel column, concentrating again, separating by a chromatographic column, and drying to obtain a sample for detection and analysis.
6. The liquid chromatography analysis method according to any one of claims 1 to 5, wherein in the analysis method, the detection limit concentration corresponding to 3 times noise is 0.37 μ g/mL.
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| CN116539743A (en) * | 2023-04-14 | 2023-08-04 | 中国农业大学 | Method for detecting specific N-nitroso compound in betel nut |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003300963A (en) * | 2002-04-09 | 2003-10-21 | Ube Ind Ltd | Method for producing 5-amino-4-nitrosopyrazole derivative |
| US20090155212A1 (en) * | 2000-03-03 | 2009-06-18 | Bratzler Robert L | Immunostimulatory nucleic acids and cancer medicament combination therapy for the treatment of cancer |
| US20090306380A1 (en) * | 2005-04-25 | 2009-12-10 | Shigeyoshi Nishino | Process for preparing 4-amino-2-alkylthio-5-pyrimidinecarbaldehyde |
| CN108037206A (en) * | 2017-12-20 | 2018-05-15 | 精晶药业股份有限公司 | A kind of efficient liquid phase detection method of Beta-alanine |
| WO2021062889A1 (en) * | 2019-09-30 | 2021-04-08 | 青岛琛蓝医药科技发展有限公司 | Method for quality control and chromatographic fingerprinting of epimedium compound product |
| CN113125606A (en) * | 2021-04-19 | 2021-07-16 | 衢州市疾病预防控制中心 | Method for simultaneously determining contents of 9N-nitrosamine compounds |
| CN116539743A (en) * | 2023-04-14 | 2023-08-04 | 中国农业大学 | Method for detecting specific N-nitroso compound in betel nut |
-
2022
- 2022-06-28 CN CN202210753532.7A patent/CN115032318B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090155212A1 (en) * | 2000-03-03 | 2009-06-18 | Bratzler Robert L | Immunostimulatory nucleic acids and cancer medicament combination therapy for the treatment of cancer |
| JP2003300963A (en) * | 2002-04-09 | 2003-10-21 | Ube Ind Ltd | Method for producing 5-amino-4-nitrosopyrazole derivative |
| US20090306380A1 (en) * | 2005-04-25 | 2009-12-10 | Shigeyoshi Nishino | Process for preparing 4-amino-2-alkylthio-5-pyrimidinecarbaldehyde |
| CN108037206A (en) * | 2017-12-20 | 2018-05-15 | 精晶药业股份有限公司 | A kind of efficient liquid phase detection method of Beta-alanine |
| WO2021062889A1 (en) * | 2019-09-30 | 2021-04-08 | 青岛琛蓝医药科技发展有限公司 | Method for quality control and chromatographic fingerprinting of epimedium compound product |
| CN113125606A (en) * | 2021-04-19 | 2021-07-16 | 衢州市疾病预防控制中心 | Method for simultaneously determining contents of 9N-nitrosamine compounds |
| CN116539743A (en) * | 2023-04-14 | 2023-08-04 | 中国农业大学 | Method for detecting specific N-nitroso compound in betel nut |
Non-Patent Citations (3)
| Title |
|---|
| B. PROKOPCZYK等: "A study of betel quid carcinogenesis IX. comparative carcinogenicity of 3-（methylnitrosamino）propionitrile and 4-（methylnitrosamino）-1-（3-pyridyl）-1-butanone upon local application to mouse skin and rat oral mucosa", CANCER LETTERS, vol. 60, 31 December 1991 (1991-12-31), pages 153 - 157 * |
| 关皓月: "1, 1, 1，2-四氟乙烷国内外质量比对研究", 1, 1, 1., vol. 24, no. 18, 31 December 2015 (2015-12-31), pages 2153 - 2156 * |
| 黄龙: "槟榔致癌物质与口腔癌", 国际口腔医学杂志, vol. 41, no. 1, 31 December 2014 (2014-12-31), pages 102 - 107 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116539743A (en) * | 2023-04-14 | 2023-08-04 | 中国农业大学 | Method for detecting specific N-nitroso compound in betel nut |
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