CN109115899A - A kind of analysis method of Fmoc amino acid - Google Patents
A kind of analysis method of Fmoc amino acid Download PDFInfo
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- CN109115899A CN109115899A CN201810632009.2A CN201810632009A CN109115899A CN 109115899 A CN109115899 A CN 109115899A CN 201810632009 A CN201810632009 A CN 201810632009A CN 109115899 A CN109115899 A CN 109115899A
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Abstract
The invention belongs to Analytical Technology of Amino Acid fields, in particular to a kind of analysis method of Fmoc amino acid, the following steps are included: step 1, Fmoc-Arg (Pbf)-OH is dissolved in acetonitrile, the concentration of Fmoc-Arg (the Pbf)-OH is 2-10mg/mL；Step 2, using (the Pbf)-OH solution of Fmoc-Arg made from step 1 as sample, efficient liquid phase chromatographic analysis is carried out；The chromatographic condition of the high performance liquid chromatography are as follows: using octadecylsilane chemically bonded silica as filler, Sinochrom ODS-BP is chromatographic column, using gradient elution, flow velocity 1.0-2.0ml/min, column temperature: room temperature, Detection wavelength 220nm, method of the invention can be to Fmoc amino acid: Fmoc-Arg (Pbf)-OH is precisely analyzed, low, the advantage that detection method is simple, stability is good, repeatability is high with high sensitivity, detection limit.
Description
Technical field
The invention belongs to Analytical Technology of Amino Acid field, in particular to a kind of analysis method of Fmoc amino acid.
Background technique
Misconnection and many side reactions easily occur for amino acid during Peptide systhesis, need to protect amino acid, synthesis
Blocking group is sloughed after polypeptide again, i.e. fluorenes methoxy carbonyl acyl group (Fmoc) is common blocking group.In the prior art, generally pass through
Fmoc is introduced with Fmoc-Cl or Fmoc-Osu, the product quality of the subsequent synthesis polypeptide of impurities affect of the Fmoc amino acid of synthesis,
Therefore, the analysis of the purity of Fmoc amino acid is of great significance.
In the prior art, the analysis of amino acid mostly uses high performance liquid chromatography (HPLC), but is different HPLC analysis item
Part produces a very large impact the analysis result accuracy of different aminoacids.
Arginine is amino acid needed by human,, must be to arginine in order to avoid the generation of by-product in Peptide systhesis
Amino and guanidine radicals protected；Fmoc-Arg (Pbf)-OH is exactly that Fmoc (9-fluorenylmethyloxycarbonyl) is utilized to protect amino, uses Pbf
(2,2,4,6,7- pentamethyl benzofuran -5- sulfonyl) protects guanidine radicals；In the prior art, Fmoc-Arg (Pbf)-OH is had no
Report.
Summary of the invention
The present invention solves the above-mentioned technical problems in the prior art, provides a kind of analysis method of Fmoc amino acid.
To solve the above problems, technical scheme is as follows:
A kind of analysis method of Fmoc amino acid, the Fmoc amino acid are Fmoc-Arg (Pbf)-OH, the analysis side
Method the following steps are included:
Step 1, Fmoc-Arg (Pbf)-OH is dissolved in acetonitrile, the concentration of Fmoc-Arg (the Pbf)-OH is 2-
10mg/mL；
Step 2, using (the Pbf)-OH solution of Fmoc-Arg made from step 1 as sample, efficient liquid phase chromatographic analysis is carried out；
The chromatographic condition of the high performance liquid chromatography are as follows:
Using octadecylsilane chemically bonded silica as filler, Sinochrom ODS-BP is chromatographic column, is washed using gradient
De-, flow velocity 1.0-2.0ml/min, column temperature: room temperature, Detection wavelength 220nm, the gradient elution are executed according to following table:
| Time/min | Mobile phase A | Mobile phase B |
| 0.01 | 50% | 50% |
| 20.0 | 0% | 100% |
Preferably, the mobile phase A is 0.1% trifluoroacetic acid solution.
Preferably, the Mobile phase B is 0.1% trifluoroacetic acid acetonitrile solution.
Preferably, the flow velocity is 1.0ml/min.
Preferably, the sample volume of the efficient liquid phase chromatographic analysis is 10 μ L.
Compared with the existing technology, advantages of the present invention is as follows,
Method of the invention can be to Fmoc amino acid: Fmoc-Arg (Pbf)-OH is precisely analyzed, and has sensitivity
High, detection limits the advantage low, detection method is simple, stability is good, repeatability is high.
Detailed description of the invention
Fig. 1 is the high-efficient liquid phase chromatogram of embodiment 1.
Specific embodiment
Embodiment 1:
A kind of analysis method of Fmoc amino acid, the Fmoc amino acid are Fmoc-Arg (Pbf)-OH, the analysis side
Method the following steps are included:
Step 1, Fmoc-Arg (Pbf)-OH is dissolved in acetonitrile, the concentration of Fmoc-Arg (the Pbf)-OH is 3mg/
mL；
Step 2, using (the Pbf)-OH solution of Fmoc-Arg made from step 1 as sample, efficient liquid phase chromatographic analysis is carried out；
The chromatographic condition of the high performance liquid chromatography are as follows:
Using octadecylsilane chemically bonded silica as filler, Sinochrom ODS-BP is chromatographic column, is washed using gradient
De-, flow velocity 1.0ml/min, column temperature: room temperature, Detection wavelength 220nm, sample volume are 10 μ L, and the gradient elution is according to following table
It executes:
| Time/min | Mobile phase A | Mobile phase B |
| 0.01 | 50% | 50% |
| 20.0 | 0% | 100% |
The mobile phase A is 0.1% trifluoroacetic acid solution；The Mobile phase B is 0.1% trifluoroacetic acid acetonitrile solution.
Embodiment 2:
A kind of analysis method of Fmoc amino acid, the Fmoc amino acid are Fmoc-Arg (Pbf)-OH, the analysis side
Method the following steps are included:
Step 1, Fmoc-Arg (Pbf)-OH is dissolved in acetonitrile, the concentration of Fmoc-Arg (the Pbf)-OH is 2mg/
mL；
Step 2, using (the Pbf)-OH solution of Fmoc-Arg made from step 1 as sample, efficient liquid phase chromatographic analysis is carried out；
The chromatographic condition of the high performance liquid chromatography are as follows:
Using octadecylsilane chemically bonded silica as filler, Sinochrom ODS-BP is chromatographic column, is washed using gradient
De-, flow velocity 2.0ml/min, column temperature: room temperature, Detection wavelength 220nm, sample volume are 10 μ L, and the gradient elution is according to following table
It executes:
| Time/min | Mobile phase A | Mobile phase B |
| 0.01 | 50% | 50% |
| 20.0 | 0% | 100% |
The mobile phase A is 0.1% trifluoroacetic acid solution；The Mobile phase B is 0.1% trifluoroacetic acid acetonitrile solution.
Embodiment 3:
A kind of analysis method of Fmoc amino acid, the Fmoc amino acid are Fmoc-Arg (Pbf)-OH, the analysis side
Method the following steps are included:
Step 1, Fmoc-Arg (Pbf)-OH is dissolved in acetonitrile, the concentration of Fmoc-Arg (the Pbf)-OH is
10mg/mL；
Step 2, using (the Pbf)-OH solution of Fmoc-Arg made from step 1 as sample, efficient liquid phase chromatographic analysis is carried out；
The chromatographic condition of the high performance liquid chromatography are as follows:
Using octadecylsilane chemically bonded silica as filler, Sinochrom ODS-BP is chromatographic column, is washed using gradient
De-, flow velocity 1.0ml/min, column temperature: room temperature, Detection wavelength 220nm, sample volume are 10 μ L, and the gradient elution is according to following table
It executes:
| Time/min | Mobile phase A | Mobile phase B |
| 0.01 | 50% | 50% |
| 20.0 | 0% | 100% |
The mobile phase A is 0.1% trifluoroacetic acid solution；The Mobile phase B is 0.1% trifluoroacetic acid acetonitrile solution.
It should be noted that above-described embodiment is only presently preferred embodiments of the present invention, there is no for the purpose of limiting the invention
Protection scope, the equivalent substitution or substitution made on the basis of the above all belong to the scope of protection of the present invention.
Claims (5)
1. a kind of analysis method of Fmoc amino acid, the Fmoc amino acid is Fmoc-Arg (Pbf)-OH, which is characterized in that institute
State analysis method the following steps are included:
Step 1, Fmoc-Arg (Pbf)-OH is dissolved in acetonitrile, the concentration of Fmoc-Arg (the Pbf)-OH is 2-10mg/
mL；
Step 2, using (the Pbf)-OH solution of Fmoc-Arg made from step 1 as sample, efficient liquid phase chromatographic analysis is carried out；
The chromatographic condition of the high performance liquid chromatography are as follows:
Using octadecylsilane chemically bonded silica as filler, Sinochrom ODS-BP is chromatographic column, using gradient elution, stream
Fast 1.0-2.0ml/min, column temperature: room temperature, Detection wavelength 220nm, the gradient elution are executed according to following table:
2. the analysis method of Fmoc amino acid as described in claim 1, which is characterized in that the mobile phase A is 0.1% trifluoro
Acetic acid solution.
3. the analysis method of Fmoc amino acid as described in claim 1, which is characterized in that the Mobile phase B is 0.1% trifluoro
Acetic acid acetonitrile solution.
4. the analysis method of Fmoc amino acid as described in claim 1, which is characterized in that the flow velocity is 1.0ml/min.
5. the analysis method of Fmoc amino acid as described in claim 1, which is characterized in that the efficient liquid phase chromatographic analysis
Sample volume is 10 μ L.
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| CN201810632009.2A CN109115899A (en) | 2018-06-19 | 2018-06-19 | A kind of analysis method of Fmoc amino acid |
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| CN201810632009.2A CN109115899A (en) | 2018-06-19 | 2018-06-19 | A kind of analysis method of Fmoc amino acid |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111505160A (en) * | 2020-05-11 | 2020-08-07 | 成都市科隆化学品有限公司 | Fmoc-protected amino acid purity and related substance analysis method |
| CN112526051A (en) * | 2020-12-18 | 2021-03-19 | 上海吉奉生物科技有限公司 | Fmoc-lysine high performance liquid chromatography determination method |
| CN112578044A (en) * | 2020-12-02 | 2021-03-30 | 成都普康生物科技有限公司 | Method for detecting Fmoc-AEEA content by high performance liquid chromatography |
| CN112946130A (en) * | 2021-02-03 | 2021-06-11 | 江苏沿海化学品检测技术服务有限公司 | Method for detecting Fmoc-Ser (tbu) -OH related substances based on high performance liquid chromatography |
| CN112986445A (en) * | 2021-03-26 | 2021-06-18 | 吉尔多肽生物制药(大连市)有限公司 | Detection method of Fmoc-Pbf-arginine related substances |
| CN112986440A (en) * | 2021-03-02 | 2021-06-18 | 吉尔多肽生物制药(大连市)有限公司 | Detection method of Fmoc-Cys (trt) -OH related substances |
| CN113092648A (en) * | 2021-04-09 | 2021-07-09 | 江苏沿海化学品检测技术服务有限公司 | Separation and purification method of amino acid |
| CN113655131A (en) * | 2021-04-23 | 2021-11-16 | 吉尔多肽生物制药(大连市)有限公司 | Fmoc-Ile-OH related substance detection method |
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| WO2004096814A1 (en) * | 2003-04-30 | 2004-11-11 | Yanaihara Institute Inc. | Polychlorobiphenyl derivatives and assay method using the same |
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| C. SCHUMANN ET AL: "Synthesis of different types of dipeptide building units containing N- or C-terminal arginine for the assembly of backbone cyclic peptides", 《J. PEPTIDE RES.》 * |
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111505160A (en) * | 2020-05-11 | 2020-08-07 | 成都市科隆化学品有限公司 | Fmoc-protected amino acid purity and related substance analysis method |
| CN111505160B (en) * | 2020-05-11 | 2021-04-06 | 成都市科隆化学品有限公司 | Fmoc-protected amino acid purity and related substance analysis method |
| CN112578044A (en) * | 2020-12-02 | 2021-03-30 | 成都普康生物科技有限公司 | Method for detecting Fmoc-AEEA content by high performance liquid chromatography |
| CN112578044B (en) * | 2020-12-02 | 2022-09-02 | 成都普康生物科技有限公司 | Method for detecting Fmoc-AEEA content by high performance liquid chromatography |
| CN112526051A (en) * | 2020-12-18 | 2021-03-19 | 上海吉奉生物科技有限公司 | Fmoc-lysine high performance liquid chromatography determination method |
| CN112526051B (en) * | 2020-12-18 | 2022-08-09 | 上海吉奉生物科技有限公司 | Fmoc-lysine high performance liquid chromatography determination method |
| CN112946130A (en) * | 2021-02-03 | 2021-06-11 | 江苏沿海化学品检测技术服务有限公司 | Method for detecting Fmoc-Ser (tbu) -OH related substances based on high performance liquid chromatography |
| CN112986440A (en) * | 2021-03-02 | 2021-06-18 | 吉尔多肽生物制药(大连市)有限公司 | Detection method of Fmoc-Cys (trt) -OH related substances |
| CN112986445A (en) * | 2021-03-26 | 2021-06-18 | 吉尔多肽生物制药(大连市)有限公司 | Detection method of Fmoc-Pbf-arginine related substances |
| CN113092648A (en) * | 2021-04-09 | 2021-07-09 | 江苏沿海化学品检测技术服务有限公司 | Separation and purification method of amino acid |
| CN113655131A (en) * | 2021-04-23 | 2021-11-16 | 吉尔多肽生物制药(大连市)有限公司 | Fmoc-Ile-OH related substance detection method |
| CN113655131B (en) * | 2021-04-23 | 2023-09-29 | 吉尔多肽生物制药(大连市)有限公司 | Fmoc-Ile-OH related substance detection method |
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