CN110297050A - A kind of discrimination method of insulin mass spectrum peptide figure and application - Google Patents

A kind of discrimination method of insulin mass spectrum peptide figure and application Download PDF

Info

Publication number
CN110297050A
CN110297050A CN201910663742.5A CN201910663742A CN110297050A CN 110297050 A CN110297050 A CN 110297050A CN 201910663742 A CN201910663742 A CN 201910663742A CN 110297050 A CN110297050 A CN 110297050A
Authority
CN
China
Prior art keywords
insulin
enzyme
peptide
injection
mass spectrum
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201910663742.5A
Other languages
Chinese (zh)
Other versions
CN110297050B (en
Inventor
白海娇
韩晓捷
伏圣青
邵建强
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Tianjin Institute Of Drug Control
Original Assignee
Tianjin Institute Of Drug Control
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Tianjin Institute Of Drug Control filed Critical Tianjin Institute Of Drug Control
Priority to CN201910663742.5A priority Critical patent/CN110297050B/en
Publication of CN110297050A publication Critical patent/CN110297050A/en
Application granted granted Critical
Publication of CN110297050B publication Critical patent/CN110297050B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • G01N30/04Preparation or injection of sample to be analysed
    • G01N30/06Preparation
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • G01N30/26Conditioning of the fluid carrier; Flow patterns
    • G01N30/28Control of physical parameters of the fluid carrier
    • G01N30/34Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • G01N30/62Detectors specially adapted therefor
    • G01N30/72Mass spectrometers
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • G01N30/88Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • G01N30/04Preparation or injection of sample to be analysed
    • G01N30/06Preparation
    • G01N2030/067Preparation by reaction, e.g. derivatising the sample
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02Column chromatography
    • G01N30/88Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
    • G01N2030/8809Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
    • G01N2030/884Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample organic compounds

Landscapes

  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
  • Peptides Or Proteins (AREA)

Abstract

The invention discloses a kind of discrimination method of insulin mass spectrum peptide figure and applications.The present invention is combined using V8 enzyme combination V8 enzyme and lysyl endopeptidase, specific for hydrolysis insulin, establishes the peptide figure containing insulin characteristic fragments by LC-MS instrument.Method is established: providing the enzymatic hydrolysis environment of pH7.5~9.5 using buffer, substrate is with enzyme by insulin: V8 enzyme: lysyl endopeptidase ≈ (100~1000): (100~500): 1 mass ratio react overnight, reaction solution dilutes 10~30 times, it is detected using liquid phase separation mass detector, establishes the feature peptide mapping of each kind insulin.Using mass spectrum peptide mapping of the invention, various different insulin can be accurately determined, method is accurate and reliable.

Description

A kind of discrimination method of insulin mass spectrum peptide figure and application
Technical field
The invention belongs to Pharmaceutical Analysis technical field, it is related to a kind of method for building up of insulin enzymatic hydrolysis mass spectrum peptide mapping and answers With.
Background technique
Insulin type recombinant product is at present for the most effective and safest treatment means of diabetes.Population base of China Greatly, diabetic's quantity is more, and insulin usage amount is big.Although recombinant technique content is high, production process is complicated, in abundant benefit Under profit driving, there are the risks that illegal retailer is mixed the spurious with the genuine using animal derived insulin.In present quality standard, for The identification of the Recombulin true and false is identified using retention time consistency, and variety classes Recombulin is intermolecular Structure and its similar, retention time is very close, practical to identify in operating process that there is certain difficulties.
Summary of the invention
The object of the present invention is to provide a kind of discrimination methods of insulin mass spectrum peptide figure.The present invention is using suitable enzymatic hydrolysis side Method is aided with Mass Spectrometer Method, and the enzymatic hydrolysis mass spectrum peptide figure for establishing each para-insulin will greatly improve the accurate reliability of identification, improves pancreas The distinguishing ability of island element provides strong technical support for the work of cracking down on the fake of insulin.
To achieve the above object, the invention discloses following technology contents:
A kind of discrimination method of insulin mass spectrum peptide figure, it is characterised in that carried out by following step:
(1) identify various para-insulins using V8 enzyme combination V8 enzyme and lysyl endopeptidase combination enzyme solution;
(2) V8 enzyme and lysyl endopeptidase combination enzyme solution identify insulin lispro;
(3) hydrolysising condition: the enzymatic hydrolysis environment of pH7.5~9.5 is provided using Tris-Hcl buffer;
(4) preparation of reaction solution: according to insulin: V8 enzyme: lysyl endopeptidase ≈ (100~1000): (100~500): 1 mass ratio prepares reaction solution respectively;
(5) chromatographic condition: using octadecylsilane chemically bonded silica as filler, using (0.1%~1%) aqueous formic acid as mobile phase A, acetonitrile are Mobile phase B, and using gradient elution, flow velocity is (0.2~0.6) ml/min, 50~60 DEG C of column temperature, mass detector;
1 eluent gradient elution requirement (volume ratio) of table
(6) measuring method: taking enzymolysis sample to inject LC-MS instrument, carries out data acquisition, measures the mass spectrogram of enzymolysis polypeptide segment.
Discrimination method of the present invention, it is characterised in that establish feature matter of a variety of insulin under two kinds of enzymatic hydrolysis conditions Peptide figure is composed, is actrapid monotard, insulin aspart, insulin glargine, paddy bad insulin, insulin lispro, pork insulin, ox pancreas islet Element etc..
The present invention uses mass detector continuous scanning within the scope of 50 ~ 2000Da, formulates the enzymatic hydrolysis matter of a variety of insulin Compose peptide figure.
According to the present invention measurement insulin mass spectrum peptide figure, part insulin characterising mass spectrometry theoretical fragment result such as the following table 2, Table 3:
2 insulin V8 enzyme hydrolysis theoretical fragment conclusive table of table
3 insulin of table hydrolyzes theoretical fragment conclusive table under the conditions of V8 enzyme and lysyl endopeptidase are combined
It includes raw material and preparation that insulin mass spectrum peptide mapping of the present invention, which is established,.Wherein Mass Spectrometer Method obtains 4 feature peptides Section.It is peptide fragment I, peptide fragment II, peptide fragment III, peptide fragment IV respectively.
The present invention further discloses the discrimination methods of insulin mass spectrum peptide figure quickly to identify the insulin true and false (example accurate Insulin aspart injection is such as pretended to be with pork insulin injection), identify insulin specific category and (passes through the feature of each insulin Mass spectrum peptide figure to insulin type realize identify) in terms of application.Experimental result is shown: other using mass spectrum peptide illustrated handbook of the invention Method can accurately identify the true and false and specific category of insulin (see attached drawing 1~4).
It is all kinds of under two kinds of enzymatic hydrolysis conditions of high spot reviews present invention mainly solves the identification of the insulin true and false and classification The stability and specificity of insulin peptide hydrolysis, main difficult point are the control of enzymatic hydrolysis condition and the inspection of feature peptide hydrolysis Survey analysis.
Possessed good effect exists the discrimination method of insulin mass spectrum peptide figure disclosed by the invention compared with prior art In:
(1) present invention establishes related special mass spectrum peptide figure for a variety of different insulin, can in insulin discrimination process Quickly and accurately to identify the true and false of insulin, while identifying the specific category of insulin.
(2) present invention will no longer compare the retention time of test sample Yu reference substance enzymolysis polypeptide segment, examine using mass spectrum After surveying device substitution ultraviolet-visible detector progress polypeptide fragment measurement, the feature matter of each insulin peptide hydrolysis will be directly relied on Spectrum peptide illustrated handbook determines each insulin.
Detailed description of the invention
Fig. 1 recombinant human insulin injection V8 enzyme hydrolysis mass spectrogram
Fig. 2 pork insulin injection V8 enzyme hydrolysis mass spectrogram
Fig. 3 recombinant human insulin injection V8 enzyme and lysyl endopeptidase combination enzymatic hydrolysis mass spectrogram
Fig. 4 Insulin Lispro Injection V8 enzyme and lysyl endopeptidase combination enzymatic hydrolysis mass spectrogram
Fig. 5 insulin aspart injection V8 enzyme hydrolysis mass spectrum peptide figure;
Fig. 6 palms off insulin aspart injection V8 enzyme hydrolysis mass spectrum peptide figure;
Fig. 7 rh-insulin's chromatogram
Fig. 8 pork insulin chromatogram.
Specific embodiment
The present invention is described below by specific embodiment.Unless stated otherwise, technological means used in the present invention It is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, it is not intended to limit the present invention Range, the spirit and scope of the invention are limited only by the claims that follow.To those skilled in the art, without departing substantially from this Under the premise of invention spirit and scope, to the various changes or change of material component and dosage progress in these embodiments It belongs to the scope of protection of the present invention.The raw materials used in the present invention and reagent are commercially available.
Tris-Hcl buffer voluntarily configure (weigh trishydroxymethylaminomethane 12.14g, add water 800ml stirring and dissolving, And it is diluted to 1000ml, pH value is adjusted with 1mol/L hydrochloric acid solution.)
Insulin source is commercially available
V8 enzyme source is commercially available
Lysyl endopeptidase source is commercially available
Experiment uses instrument for UPLC-XEVO G2-XsQTof LC-MS instrument, is equipped with ESI ion source.
Embodiment 1
Rh-insulin, insulin aspart, insulin lispro and pork insulin injection V8 enzyme hydrolysis mass spectrum peptide figure and V8 enzyme With lysyl endopeptidase combination mass spectrum peptide figure measurement
Respectively take injection liquid samples 1 respectively, be transferred to 15ml centrifuge tube, be suitable for Tris-Hcl buffer adjust sample pH to 8.5, dilution volume to 15ml.
(1) 600 μ l of dilute sample is taken, 200 μ l(1mg/ml of V8 enzyme is added), then 400 μ l Tris- are added in ambient temperature overnight Hcl buffer;
(2) 600 μ l of dilute sample is taken, 200 μ l(1mg/ml of V8 enzyme is added) and 50 μ l(20 μ g/ml of lysyl endopeptidase), Then 400 μ l Tris-Hcl buffers are added in ambient temperature overnight.Sample dilutes 20 times before sample introduction.Using Waters ACQUITY UPLC®CSHTMC18,1.7 μm, 50 × 2.1mm(I.D) chromatographic column according to following table gradient condition is with 0.1% aqueous formic acid Mobile phase A, acetonitrile are Mobile phase B, 0.4ml/min flow velocity, 60 DEG C of column temperatures, 5 μ l of sample introduction measurement, acquisition ion flow graph;
4 eluent gradient elution requirement (volume ratio) of table
Four characteristic polypeptide fragments that sample digests generation under two kinds of enzymatic hydrolysis conditions are shown in Table 5 and table 6:
54 kinds of insulin V8 enzyme hydrolysis peptide segment tables of table
64 kinds of insulin V8 enzymes of table and lysyl endopeptidase are combined range of hydrolysed peptides segment table
Under the conditions of V8 enzyme and V8 enzyme and lysyl endopeptidase are combined, according to the feature for digesting generation under two kinds of enzymatic hydrolysis conditions Property peptide fragment can accurately identify the true and false and the specific classification of insulin of various insulin.
Embodiment 2
Rh-insulin and the combination enzymatic hydrolysis mass spectrum peptide figure measurement of Insulin Lispro Injection V8 enzyme and lysyl endopeptidase
Injection liquid samples 1 is respectively taken, 15ml centrifuge tube is transferred to, it is dilute to be suitable for Tris-Hcl buffer adjusting sample pH to 9 Volume is released to 15ml.1000 μ l of dilute sample is taken, 100 μ l(1mg/ml of V8 enzyme is added) and 20 μ l(20 μ of lysyl endopeptidase G/ml), then 400 μ l Tris-Hcl buffers are added in ambient temperature overnight.Sample dilutes 10 times before sample introduction.According to 1 ladder of embodiment The measurement of degree condition, using 0.7% aqueous formic acid as mobile phase A, acetonitrile is Mobile phase B, 0.2ml/min flow velocity, 55 DEG C of column temperatures, into 5 μ l of sample acquires ion flow graph;The four characteristic polypeptide fragments generated after sample enzymatic hydrolysis are shown in Table 7:
7 rh-insulin of table and insulin lispro V8 enzyme and lysyl endopeptidase are combined Peptides segment table
Under V8 enzyme and V8 enzyme and lysyl endopeptidase combination enzymatic hydrolysis condition, rh-insulin and insulin lispro can be with Effectively identified and is distinguished.
Embodiment 3
Insulin aspart injection and personation insulin aspart injection V8 enzyme hydrolysis mass spectrum peptide figure measurement
Injection liquid samples 1 is respectively taken, 15ml centrifuge tube is transferred to, sample pH is adjusted to 7.5 with Tris-Hcl buffer, is diluted Volume is to 15ml.1000 μ l of dilute sample is taken, 200 μ l(1mg/ml of V8 enzyme is added), then 400 μ l are added in ambient temperature overnight Tris-Hcl buffer.Sample dilutes 30 times before sample introduction.It is flowing with 0.5% aqueous formic acid according to 1 gradient condition of embodiment Phase A, acetonitrile are Mobile phase B, 0.6ml/min flow velocity, 50 DEG C of column temperatures, 5 μ l of sample introduction measurement, acquisition ion flow graph.After sample enzymatic hydrolysis The four characteristic polypeptide fragments generated are shown in Table 8:
8 insulin aspart of table and personation insulin aspart V8 enzyme Peptides segment table
Under V8 enzyme enzymatic hydrolysis condition, insulin aspart and the available effective identification of personation insulin aspart and differentiation.
Recombinant human insulin injection and pork insulin injection chromatogram under 4 liquid phase chromatogram condition of embodiment
One, sample is respectively taken, and each accurate measurement is appropriate, and it is single that about insulin-containing 10 is made in every 1ml with 0.01mol/L hydrochloric acid solution Position solution, using C18 chromatographic column, with 0.2mol/L sulfate buffer (pH2.3)-acetonitrile (74:26) for mobile phase, 1.0ml/min flow velocity detects under the conditions of 214nm Detection wavelength.Rh-insulin's chromatogram and pork insulin chromatogram are shown in The following figure Fig. 7 and Fig. 8.
Through embodiment 4 as can be seen that under identical chromatographic conditions, rh-insulin and pork insulin retention time ratio It is closer to, by embodiment 1 to embodiment 3 it can be seen that the foundation of enzymatic hydrolysis characterising mass spectrometry peptide figure can be identified accurately and effectively The true and false of insulin and the specific category of insulin, this method avoid due to insulin molecule amount it is close, in identical chromatography Under the conditions of variety classes insulin retention time it is close, thus a possibility that bringing mistake for result judgement, the side of substantially increasing The specificity of method.

Claims (5)

1. a kind of discrimination method of insulin mass spectrum peptide figure, it is characterised in that carried out by following step:
(1) identify various para-insulins using V8 enzyme combination V8 enzyme and lysyl endopeptidase combination enzyme solution;
(2) V8 enzyme and lysyl endopeptidase combination enzyme solution identify insulin lispro;
(2) hydrolysising condition: using the enzymatic hydrolysis environment of pH of buffer 7.5~9.5;The buffer refers to that Tris-Hcl is buffered Liquid;
(3) preparation of reaction solution: according to insulin: V8 enzyme: lysyl endopeptidase ≈ (100~1000): (100~500): 1 mass ratio prepares reaction solution respectively;
(4) chromatographic condition: being stream with 0.1%~1% aqueous formic acid of volume ratio using octadecylsilane chemically bonded silica as filler Dynamic phase A, acetonitrile is Mobile phase B, and using gradient elution, flow velocity is 0.2~0.6ml/min, 50~60 DEG C of column temperature, Mass Spectrometer Method Device;
(5) chromatography gradient condition is as follows:
1 eluent gradient elution requirement (volume ratio) of table
(6) measuring method: taking enzymolysis sample to inject LC-MS instrument, carries out data acquisition, measures the mass spectrogram of enzymolysis polypeptide segment.
2. discrimination method described in claims 1, it is characterised in that feature matter of a variety of insulin under two kinds of enzymatic hydrolysis conditions Peptide figure is composed, is actrapid monotard, insulin aspart, insulin glargine, paddy bad insulin, insulin lispro, pork insulin, ox pancreas islet Element.
3. discrimination method described in claims 1, it is characterised in that it includes raw material and preparation that insulin mass spectrum peptide mapping, which is established,; The raw material and preparation refers to: recombinant insulinum primary material, biosynthetic human insulin injection, protamine biosynthesis Human insulin injection, protamine biosynthetic human insulin injection (premix 30R), protamine biosynthetic human insulin note Penetrate liquid (premix 50R), insulin aspart raw material, insulin aspart injection, 30 injection of insulin aspart, insulin aspart 50 Injection.
4. discrimination method described in claims 1, it is characterised in that Mass Spectrometer Method obtains feature peptide fragment.
5. the discrimination method of insulin mass spectrum peptide figure described in claims 1 quickly identifies the insulin true and false accurate, identify pancreas Application in terms of the element specific category of island.
CN201910663742.5A 2019-07-23 2019-07-23 Identification method and application of insulin mass spectrum peptide map Active CN110297050B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201910663742.5A CN110297050B (en) 2019-07-23 2019-07-23 Identification method and application of insulin mass spectrum peptide map

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201910663742.5A CN110297050B (en) 2019-07-23 2019-07-23 Identification method and application of insulin mass spectrum peptide map

Publications (2)

Publication Number Publication Date
CN110297050A true CN110297050A (en) 2019-10-01
CN110297050B CN110297050B (en) 2022-04-12

Family

ID=68031625

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201910663742.5A Active CN110297050B (en) 2019-07-23 2019-07-23 Identification method and application of insulin mass spectrum peptide map

Country Status (1)

Country Link
CN (1) CN110297050B (en)

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111351880A (en) * 2020-04-01 2020-06-30 上海中科新生命生物科技有限公司 Mass spectrum detection method for recombinant human insulin biological analysis based on immunoaffinity
CN111778231A (en) * 2020-07-29 2020-10-16 珠海冀百康生物科技有限公司 Purification method of lysyl endopeptidase
CN111948315A (en) * 2020-08-13 2020-11-17 中国科学院长春应用化学研究所 Sequence analysis method and identification method of insulin or insulin analogue
CN112763636A (en) * 2020-12-07 2021-05-07 佛山汉腾生物科技有限公司 Peptide map analysis method
CN115808484A (en) * 2023-01-18 2023-03-17 北京惠之衡生物科技有限公司 Method for detecting residual lysyl endonuclease in insulin or insulin analogue
CN116046947A (en) * 2023-01-28 2023-05-02 北京惠之衡生物科技有限公司 Method for detecting residual lysyl endonuclease in insulin diglucer
CN116908321A (en) * 2023-06-21 2023-10-20 南京汉欣医药科技有限公司 Protamine sulfate peptide map detection method
CN116908321B (en) * 2023-06-21 2024-05-31 南京汉欣医药科技有限公司 Protamine sulfate peptide map detection method

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103392219A (en) * 2010-12-28 2013-11-13 探索诊断投资公司 Quantitation of insulin by mass spectrometry
CN105301125A (en) * 2015-09-11 2016-02-03 东阿阿胶股份有限公司 Method for identifying main deamination product site of recombinant human insulin or similar matter thereof
CN108896682A (en) * 2018-07-18 2018-11-27 杭州汇健科技有限公司 A kind of quick mass spectral analysis of peptide fingerprinting spectrum and spectrogram method of discrimination

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103392219A (en) * 2010-12-28 2013-11-13 探索诊断投资公司 Quantitation of insulin by mass spectrometry
CN105301125A (en) * 2015-09-11 2016-02-03 东阿阿胶股份有限公司 Method for identifying main deamination product site of recombinant human insulin or similar matter thereof
CN108896682A (en) * 2018-07-18 2018-11-27 杭州汇健科技有限公司 A kind of quick mass spectral analysis of peptide fingerprinting spectrum and spectrogram method of discrimination

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
JIA-CHUAN HSU ET AL.: "Identification of Recombinant Insulin Analogues by Peptide Mapping Method", 《JOURNAL OF FOOD AND DRUG ANALYSIS》 *
张培培 等: "胰岛素的HPLC肽图谱分析", 《药物生物技术》 *

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111351880A (en) * 2020-04-01 2020-06-30 上海中科新生命生物科技有限公司 Mass spectrum detection method for recombinant human insulin biological analysis based on immunoaffinity
CN111778231A (en) * 2020-07-29 2020-10-16 珠海冀百康生物科技有限公司 Purification method of lysyl endopeptidase
CN111948315A (en) * 2020-08-13 2020-11-17 中国科学院长春应用化学研究所 Sequence analysis method and identification method of insulin or insulin analogue
CN112763636A (en) * 2020-12-07 2021-05-07 佛山汉腾生物科技有限公司 Peptide map analysis method
CN112763636B (en) * 2020-12-07 2022-04-19 佛山汉腾生物科技有限公司 Peptide map analysis method
CN115808484A (en) * 2023-01-18 2023-03-17 北京惠之衡生物科技有限公司 Method for detecting residual lysyl endonuclease in insulin or insulin analogue
CN115808484B (en) * 2023-01-18 2023-04-14 北京惠之衡生物科技有限公司 Method for detecting residual lysyl endonuclease in insulin or insulin analogue
CN116046947A (en) * 2023-01-28 2023-05-02 北京惠之衡生物科技有限公司 Method for detecting residual lysyl endonuclease in insulin diglucer
CN116046947B (en) * 2023-01-28 2023-06-02 北京惠之衡生物科技有限公司 Method for detecting residual lysyl endonuclease in insulin diglucer
CN116908321A (en) * 2023-06-21 2023-10-20 南京汉欣医药科技有限公司 Protamine sulfate peptide map detection method
CN116908321B (en) * 2023-06-21 2024-05-31 南京汉欣医药科技有限公司 Protamine sulfate peptide map detection method

Also Published As

Publication number Publication date
CN110297050B (en) 2022-04-12

Similar Documents

Publication Publication Date Title
CN110297050A (en) A kind of discrimination method of insulin mass spectrum peptide figure and application
Jeppsson et al. Toward standardization of carbohydrate-deficient transferrin (CDT) measurements: I. Analyte definition and proposal of a candidate reference method
Kahsai et al. Monitoring protein conformational changes and dynamics using stable-isotope labeling and mass spectrometry
CN111896651B (en) Agkistrodon halys venom thrombin-like enzyme characteristic polypeptide and application thereof
US7807401B2 (en) Reagent for digestion of hemoglobin
CN110736802B (en) Method for quantitatively detecting α -glucosidase in honey by liquid chromatography-tandem mass spectrometry
CN110045028A (en) A kind of Preclinical metabolism and pharmacokinetics study mass spectrometric analysis method of Avastin
Vainauskas et al. Profiling of core fucosylated N-glycans using a novel bacterial lectin that specifically recognizes α1, 6 fucosylated chitobiose
Klammt et al. Functional analysis of cell‐free‐produced human endothelin B receptor reveals transmembrane segment 1 as an essential area for ET‐1 binding and homodimer formation
Thevis et al. New drugs and methods of doping and manipulation
Fang et al. Comparative analysis of intact glycopeptides from mannose receptor among different breast cancer subtypes using mass spectrometry
CN106872591A (en) A kind of composition, kit and its detection method identified for authenticity of hide glue
CN106153785A (en) A kind of online sample introduction of aflatoxin analyzes method
CN110531015A (en) The method and its application of infliximab concentration in a kind of detection serum
CN106841497A (en) For detecting colla carapacis et plastri testudinis, deer horn glue, donkey-hide gelatin and its composition of donkey hide derived components content, kit and detection method in adulterant glue
Van Der Gugten et al. Quantitation of insulin analogues in serum using immunoaffinity extraction, liquid chromatography, and tandem mass spectrometry
CN111189940B (en) Method for detecting soluble epoxide hydrolase by liquid chromatography-mass spectrometry
CN110498838B (en) Characteristic peptide segment for detecting FPGS (planar-repeats-GS) and GGH (GGH) protein expression level and application thereof
CN102175804A (en) Absolute quantitative method for biomass spectra of CYP450 enzyme hypotypes
Olivares et al. Reactivity of different LH and FSH standards and preparations in the world health organization matched reagents for enzyme-linked immunoassays of gonadotrophins
CN106841437A (en) Composition, kit and its detection method for detecting donkey hide derived components content in donkey-hide gelatin
CN108333287A (en) A kind of quantitative detecting method of Cornu Saigae Tataricae powder
CN109358131B (en) The rapid analysis method of monoclonal antibody N sugar sugar-type
CN111983043B (en) Method for detecting poloxamer residual quantity in recombinant human urokinase raw material for injection
EP3405785A1 (en) Bio-analytical method for insulin analogues

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant