WO2023082512A1 - Polypeptide marqueur de ver à soie rigide et procédé d'identification de vers à soie rigides, de produits d'extrait aqueux de ver à soie rigide et d'autres produits de ver à soie rigide - Google Patents

Polypeptide marqueur de ver à soie rigide et procédé d'identification de vers à soie rigides, de produits d'extrait aqueux de ver à soie rigide et d'autres produits de ver à soie rigide Download PDF

Info

Publication number
WO2023082512A1
WO2023082512A1 PCT/CN2022/080707 CN2022080707W WO2023082512A1 WO 2023082512 A1 WO2023082512 A1 WO 2023082512A1 CN 2022080707 W CN2022080707 W CN 2022080707W WO 2023082512 A1 WO2023082512 A1 WO 2023082512A1
Authority
WO
WIPO (PCT)
Prior art keywords
silkworm
products
stiff
silkworms
mobile phase
Prior art date
Application number
PCT/CN2022/080707
Other languages
English (en)
Chinese (zh)
Inventor
李国卫
胡绮萍
何民友
童培珍
邱韵静
程学仁
魏梅
曾荟
陈向东
Original Assignee
广东一方制药有限公司
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 广东一方制药有限公司 filed Critical 广东一方制药有限公司
Publication of WO2023082512A1 publication Critical patent/WO2023082512A1/fr

Links

Images

Classifications

    • 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
    • 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
    • G01N30/62—Detectors specially adapted therefor
    • G01N30/72—Mass spectrometers
    • 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
    • G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
    • G01N30/8679—Target compound analysis, i.e. whereby a limited number of peaks is analysed
    • 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
    • G01N2030/065—Preparation using different phases to separate parts of sample
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the amino acid sequence is the peptide segment shown in SEQ ID No.1, the m/z of the parent ion is 823.36, and the m/z of the product ions are 1070.47 and 1345.57.
  • a method for identifying the silkworm, silkworm water extract products and other silkworm products includes the step of detecting the characteristic components of the silkworm in the sample to be identified, and the characteristic components of the silkworm are the described Bombyx mori characteristic polypeptide.
  • Fig. 9 is the MRM mass spectrogram (m/ z 823);
  • Fig. 11 is the MRM mass spectrogram (m/z 823) of the prescription preparation containing silkworm;
  • Fig. 12 is the MRM mass spectrogram (m/z 637) of the prescription preparation containing silkworm.
  • the technical features described in open form include closed technical solutions consisting of the enumerated features, as well as open technical solutions including the enumerated features.
  • Technical scheme of the present invention comprises:
  • the present invention provides a method for identifying the silkworm, the water extract product of the silkworm and other silkworm products, the identification method includes the step of detecting the characteristic components of the silkworm in the sample to be identified, and the silkworm
  • the characteristic component is the above-mentioned characteristic polypeptide of Bombyx mori.
  • the gradient elution conditions include: 0min-3min, the volume percentage of the mobile phase A is 3%; 3min-8min, the volume percentage of the mobile phase A increases from 3% to 5%; 8min ⁇ 10min, the volume percentage of the mobile phase A is 5%; 10min ⁇ 18min, the volume percentage of the mobile phase A rises from 5% to 7%; 18min ⁇ 19min, the volume percentage of the mobile phase A increases from 7% Increase to 90%; 19min-21min, the volume percentage of the mobile phase A is 90%.
  • Adopting the above-mentioned detection conditions has the following advantages: shortening the detection time and saving the detection cost; optimization of the mass spectrometry conditions increases its response, etc.
  • suitable preparation conditions can be selected according to the type of the sample to be identified, such as a suitable extraction method (for example, the silkworm medicinal material and decoction pieces can adopt the method of reflux extraction, The standard decoction of silkworm and the standard decoction of fried silkworm can adopt the mode of ultrasonic extraction, etc.), an appropriate amount of trypsin.
  • a suitable extraction method for example, the silkworm medicinal material and decoction pieces can adopt the method of reflux extraction, The standard decoction of silkworm and the standard decoction of fried silkworm can adopt the mode of ultrasonic extraction, etc.
  • the other silkworm products include, but are not limited to: Zhongfeng Huichun Pills, Rupi Sanjie Capsules, Yixian Pills and Xiaoer Qizhen Pills. It should be noted that there is no sequence limitation for the steps involved in the identification method of the present invention.
  • Transfer the decoction to a round-bottomed flask use a rotary evaporator to concentrate under reduced pressure and low temperature (temperature: 65°C; vacuum degree: -0.08MPa ⁇ -0.1MPa), rotate at a speed of 50 ⁇ 90 rpm, and concentrate to a volume of about 100ml ; Under magnetic stirring, precisely absorb 2ml of the liquid extract and evenly distribute it in 10ml vials, transfer to a vacuum freeze dryer to freeze-dry, take it out, and roll an aluminum cap to get it.
  • test solution (1) Refer to the following methods to prepare silkworm medicinal materials, silkworm decoction pieces, fried silkworm decoction pieces, chafer medicinal materials, mealworm medicinal materials, tortoise shell medicinal materials, gallinaceous gold medicinal materials, leech medicinal materials, cicada slough medicinal materials, earthworm medicinal materials, and wood beetle medicinal materials.
  • Chromatographic conditions Chromatographic column: Phenomenes Titank C18 chromatographic column (1.7 ⁇ m, 2.1mm ⁇ 150mm); mobile phase: acetonitrile-0.1% formic acid aqueous solution, gradient elution according to the table below; column temperature: 30°C; flow rate: 0.30 mL/min; injection volume: 5 ⁇ L.
  • test solution was injected at 0, 2, 5, 8, 10, and 12 hours respectively, and the RSD values of the two pairs of characteristic ion pair peak areas were in the range of 1.27% to 4.41%, both less than 5%, indicating that the test product The solution has good stability within 12 hours.
  • 2# chromatographic column is Waters BEH C18 chromatographic column
  • 3# chromatographic column is Shimadzu Shim-pack C18-AQ chromatographic column
  • test solution Take an appropriate amount of fried silkworm Chinese medicine formula granules, prepare the test solution according to the preparation method of the test solution determined under "1 Preparation of the test solution", accurately draw 5 ⁇ l and inject it into the liquid mass spectrometer, press “3 Chromatography and Mass spectrometry conditions” under the conditions for determination, the experimental results are shown in Table 11.

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)
  • Engineering & Computer Science (AREA)
  • Library & Information Science (AREA)
  • Peptides Or Proteins (AREA)
  • Other Investigation Or Analysis Of Materials By Electrical Means (AREA)

Abstract

L'invention concerne un polypeptide marqueur de ver à soie rigide et un procédé d'identification de vers à soie rigides, de produits d'extrait aqueux de ver à soie rigide et d'autres produits de ver à soie rigide. Le polypeptide marqueur de ver à soie rigide comprend un fragment peptidique ayant une séquence d'acides aminés telle que représentée dans SEQ ID No 1 et/ou un fragment peptidique ayant une séquence d'acides aminés telle que représentée dans SEQ ID No 2. Le procédé d'identification de vers à soie rigides, des produits d'extrait aqueux de ver à soie rigide et d'autres produits de ver à soie rigide comprend une étape d'utilisation d'un procédé de spectrométrie de masse de chromatographie liquide à ultra-haute performance pour détecter des composants de marqueur de ver à soie rigide dans un échantillon à identifier. Selon le procédé, le polypeptide marqueur de ver à soie rigide dans l'échantillon est détecté pour identifier des vers à soie rigides et des produits de ceux-ci, ce qui permet de résoudre le problème de difficulté d'identification de décoction classique de médecine chinoise traditionnelle, des granulés de formule de médecine chinoise traditionnelle, ainsi que la formulation et la préparation à partir de vers à soie rigides en raison de caractères d'apparence manquants et la fourniture d'une base pour le contrôle de qualité et l'évaluation de vers à soie rigides et de produits de ceux-ci, en particulier des produits d'extrait aqueux de ver à soie rigide.
PCT/CN2022/080707 2021-11-10 2022-03-14 Polypeptide marqueur de ver à soie rigide et procédé d'identification de vers à soie rigides, de produits d'extrait aqueux de ver à soie rigide et d'autres produits de ver à soie rigide WO2023082512A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202111327905.6 2021-11-10
CN202111327905.6A CN114113376B (zh) 2021-11-10 2021-11-10 僵蚕特征多肽以及僵蚕、僵蚕水提物制品和其他僵蚕制品的鉴别方法

Publications (1)

Publication Number Publication Date
WO2023082512A1 true WO2023082512A1 (fr) 2023-05-19

Family

ID=80378016

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2022/080707 WO2023082512A1 (fr) 2021-11-10 2022-03-14 Polypeptide marqueur de ver à soie rigide et procédé d'identification de vers à soie rigides, de produits d'extrait aqueux de ver à soie rigide et d'autres produits de ver à soie rigide

Country Status (2)

Country Link
CN (1) CN114113376B (fr)
WO (1) WO2023082512A1 (fr)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114113376B (zh) * 2021-11-10 2023-08-29 广东一方制药有限公司 僵蚕特征多肽以及僵蚕、僵蚕水提物制品和其他僵蚕制品的鉴别方法
CN115290782A (zh) * 2022-08-01 2022-11-04 北京振东光明药物研究院有限公司 含水蛭、全蝎、僵蚕和/或土鳖虫的制剂的鉴别方法及其应用

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105683373A (zh) * 2013-10-25 2016-06-15 国立研究开发法人农业生物资源研究所 外源性基因表达增强剂
CN106048020A (zh) * 2016-06-15 2016-10-26 中国医学科学院药用植物研究所 动物药材僵蚕的基因身份证
CN107164471A (zh) * 2017-05-18 2017-09-15 华南农业大学 一种快速鉴别中药材僵蚕中白僵菌真伪的分子鉴定方法
US20180011107A1 (en) * 2016-07-07 2018-01-11 Genysis Lab Inc. Identification of the presence of specific polypeptides by liquid chromatography and mass spectrometry
US9933416B1 (en) * 2013-07-30 2018-04-03 Pioneer Hi-Bred International, Inc. Detection and quantification of polypeptides in plants without a reference standard by mass spectrometry
CN109521132A (zh) * 2018-11-16 2019-03-26 广东省农业科学院蚕业与农产品加工研究所 一种僵蚕hplc指纹图谱的建立方法
CN109521110A (zh) * 2018-11-16 2019-03-26 广东省农业科学院蚕业与农产品加工研究所 一种基于hplc指纹图谱评价僵蚕僵化程度的方法
CN110824032A (zh) * 2019-09-26 2020-02-21 成都中医药大学 一种僵蚕及其伪品的指纹图谱鉴别方法
CN114113376A (zh) * 2021-11-10 2022-03-01 广东一方制药有限公司 僵蚕特征多肽以及僵蚕、僵蚕水提物制品和其他僵蚕制品的鉴别方法

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107118266A (zh) * 2017-04-13 2017-09-01 上海交通大学 一种具有α‑糖苷酶抑制活性的丝胶肽及其制备与应用

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9933416B1 (en) * 2013-07-30 2018-04-03 Pioneer Hi-Bred International, Inc. Detection and quantification of polypeptides in plants without a reference standard by mass spectrometry
CN105683373A (zh) * 2013-10-25 2016-06-15 国立研究开发法人农业生物资源研究所 外源性基因表达增强剂
CN106048020A (zh) * 2016-06-15 2016-10-26 中国医学科学院药用植物研究所 动物药材僵蚕的基因身份证
US20180011107A1 (en) * 2016-07-07 2018-01-11 Genysis Lab Inc. Identification of the presence of specific polypeptides by liquid chromatography and mass spectrometry
CN107164471A (zh) * 2017-05-18 2017-09-15 华南农业大学 一种快速鉴别中药材僵蚕中白僵菌真伪的分子鉴定方法
CN109521132A (zh) * 2018-11-16 2019-03-26 广东省农业科学院蚕业与农产品加工研究所 一种僵蚕hplc指纹图谱的建立方法
CN109521110A (zh) * 2018-11-16 2019-03-26 广东省农业科学院蚕业与农产品加工研究所 一种基于hplc指纹图谱评价僵蚕僵化程度的方法
CN110824032A (zh) * 2019-09-26 2020-02-21 成都中医药大学 一种僵蚕及其伪品的指纹图谱鉴别方法
CN114113376A (zh) * 2021-11-10 2022-03-01 广东一方制药有限公司 僵蚕特征多肽以及僵蚕、僵蚕水提物制品和其他僵蚕制品的鉴别方法

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
BAI YUNPENG; ZHAO QIANG; HE MEIXI; YE XUETING; ZHANG XIAOZHE: "Extensive characterization and differential analysis of endogenous peptides fromBombyx batryticatususing mass spectrometric approach", JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS, ELSEVIER B.V., AMSTERDAM, NL, vol. 163, 18 September 2018 (2018-09-18), AMSTERDAM, NL , pages 78 - 87, XP085515552, ISSN: 0731-7085, DOI: 10.1016/j.jpba.2018.09.033 *
CHENG XIAN-LONG; CHEN JIA; LI MING-HUA; ZHANG QIAN-QIAN; LIU WEI; WEI FENG; MA SHUANG-CHENG: "Identification Study of Glue Medicines", CHINESE PHARMACEUTICAL JOURNAL, vol. 50, no. 02, 22 January 2015 (2015-01-22), pages 104 - 108, XP009545636, ISSN: 1001-2494, DOI: 10.11669/cpj.2015.02.003 *
DATABASE Protein 02-12-2020, ANONYMOUS : "sericin 1 isoform X1 [Bombyx mori]", XP093067180, retrieved from NCBI Database accession no. XP_037869538.1 *
LIU YUWEN; ZHI YANG; YU CHEN; ZHILING LU: "Identification Study of Trionycis-Shell Glue by UPLC-MS/MS Marker Peptide", CHINESE JOURNAL OF MODERN APPLIED PHARMACY,20200229CHINESE PHARMACEUTICAL ASSOCIATION, CN, vol. 30, no. 04, 25 February 2020 (2020-02-25), pages 3061 - 3063, XP055922069, ISSN: 1007-7693 *

Also Published As

Publication number Publication date
CN114113376B (zh) 2023-08-29
CN114113376A (zh) 2022-03-01

Similar Documents

Publication Publication Date Title
WO2023082512A1 (fr) Polypeptide marqueur de ver à soie rigide et procédé d'identification de vers à soie rigides, de produits d'extrait aqueux de ver à soie rigide et d'autres produits de ver à soie rigide
Cox et al. The utility of metabolomics in natural product and biomarker characterization
CN103760269B (zh) 一种兽药残留的检测方法
Park et al. Metabolomic approach for discrimination of processed ginseng genus (Panax ginseng and Panax quinquefolius) using UPLC-QTOF MS
CN105572212A (zh) 一种基于可视化质谱信息的生晒参和红参快速识别方法
CN116284247B (zh) 一种鉴别鹿角胶混淆品的特征多肽及其组合方法、应用
Kohler et al. The rise of hydrophilic interaction chromatography in untargeted clinical metabolomics
Hu et al. Determination and analysis of monosaccharides in Polygonatum cyrtonema Hua polysaccharides from different areas by ultra‐high‐performance liquid chromatography quadrupole trap tandem mass spectrometry
Guo et al. Identification of velvet antler and its mixed varieties by UPLC-QTOF-MS combined with principal component analysis
WO2022078020A1 (fr) Procédé de criblage de combinaison de polypeptides caractéristiques à des fins de détection de carapace et d'écaille de tortue, et son application
CN114113381B (zh) 一种舒氏海龙特征多肽及其应用和鉴别舒适海龙的方法
CN113896770B (zh) 一种鳖甲特征肽段及其检测方法
CN109374785B (zh) 淡竹叶药材uplc特征图谱的构建方法和检测方法
Qi et al. Rapid screening of illegal additives in functional food using desorption electrospray ionization mass spectrometry imaging
CN109100461B (zh) 一种利用蛋白组学技术区分有机大米和非有机大米的方法
CN110596277B (zh) 一种女贞子生品和酒制品的鉴别方法
Hui et al. A novel approach to characterize chemical consistency of traditional Chinese medicine Fuzi Lizhong pills by GC-MS and RRLC-Q-TOFMS
Cheng et al. Identification of four cornua by ultra‐performance liquid chromatography with time‐of‐flight mass spectrometry coupled with principal component analysis
CN107607654B (zh) 一种核桃花中黄酮类化学成分的分析方法
WO2022016809A1 (fr) Polypeptide spécifique d'endothelium corneum gigeriae galli et son application dans l'identification d'endothelium corneum gigeriae galli
CN115541756A (zh) 车前科中药药材或制剂的指纹图谱的构建方法、检测方法和鉴别方法
CN113759011B (zh) 一种银柴胡及其制剂特征图谱的建立方法
CN114563497A (zh) 一种小柴胡胶囊成分的定量指纹图谱检测方法
CN107037139B (zh) 免疫亲和柱净化-超高效液相色谱-串联质谱检测鱼虾中3-甲基-喹噁啉-2-羧酸的方法
CN114395014B (zh) 一种牛角特征肽段及其检测方法

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 22891324

Country of ref document: EP

Kind code of ref document: A1