Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for measuring multiple types of ATP (adenosine triphosphate) associated products in aquatic product with high performance liquid chromatography-diode array method

A high-performance liquid chromatography and diode array technology, applied in the field of analytical chemistry, can solve the problems of qualitative column contamination, poor separation effect, and limited types, and meet the requirements of simple equipment and analysis conditions, improved detection efficiency, and less contaminated columns. Effect

Active Publication Date: 2014-12-24
SHANGHAI OCEAN UNIV
View PDF5 Cites 15 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a high performance liquid chromatography-diode array method that is simple to operate, fast in separation speed, does not use ion-pair reagents, and is quantitatively and qualitatively accurate, and can simultaneously measure multiple ATP-related products in aquatic products, so as to solve the above prior art However, there are deficiencies such as poor separation effect, limited species, long analysis time, serious column contamination and inaccurate qualitative analysis.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for measuring multiple types of ATP (adenosine triphosphate) associated products in aquatic product with high performance liquid chromatography-diode array method
  • Method for measuring multiple types of ATP (adenosine triphosphate) associated products in aquatic product with high performance liquid chromatography-diode array method
  • Method for measuring multiple types of ATP (adenosine triphosphate) associated products in aquatic product with high performance liquid chromatography-diode array method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1 Separation and detection of 10 kinds of ATP related substances

[0057] (1) Preparation of standard sample solution: Use dipotassium hydrogen phosphate buffer solution (mobile phase B phase) with pH=5.9 and concentration of 15mmol / L as the solvent, and weigh 10 kinds of ATP related substance standard products ATP, GMP, IMP respectively , ADP, Hx, AMP, Ad, HxR, AdR and Xt, and prepare a series of standard solutions of ATP related substances with concentrations of 0.01-750 μg / mL.

[0058] (2) Determination of the optimal UV detection wavelength: take a standard solution with a concentration of 100 μg / mL, and use a diode array detector to scan to determine the UV absorption spectrum and optimal detection wavelength (UV Maximum absorption wavelength), determine the best UV detection wavelengths of ATP, GMP, IMP, ADP, Hx, AMP, Ad, HxR, AdR and Xt are 259.1, 253.2, 248.4, 259.1, 248.4, 259.1, 260.3, 248.4, 259.1 , 267.4nm, the result figure 1 .

[0059] (3) Prepa...

Embodiment 2

[0077] Embodiment 2 measures aquatic product (1)

[0078] The aquatic products tested include Penaeus monodon and Chinese mitten crab.

[0079] Preparation of the solution to be tested: Take 5.0 g of fresh aquatic products (shrimp meat or crab meat) in a centrifuge tube, add 10 mL of 10% perchloric acid pre-cooled to 4 °C to homogenate for 1 min, stir evenly with a glass rod, and place in the freezer Centrifuge at 10000rpm at 4°C for 10min, take the supernatant and filter, add the remaining precipitate to the pre-cooled 5% perchloric acid solution for leaching and centrifugation, repeat 2 times, combine the supernatant, and use KOH The solution neutralized the supernatant to a pH of 5.9. Potassium perchlorate was removed by filtration, and dipotassium hydrogen phosphate buffer solution (phase B of the mobile phase) with a pH of 5.9 and a concentration of 15 mmol / L was added to a 50 mL volumetric flask.

[0080]Take 2 mL of the above sample solution, filter it through a water...

Embodiment 3

[0085] Take 10 kinds of ATP-related standard substances such as ATP, GMP, IMP, ADP, Hx, Xt, AMP, Ad, HxR and AdR, and use pH=5.9 and a concentration of 15mmol / L dipotassium hydrogen phosphate buffer to make a mixed solution, The concentration of each ATP conjugate was 100 μg / mL.

[0086] The above mixed solution was detected by Waters e2695 high performance liquid chromatography (Waters, USA) (equipped with Waters 2996 diode array detector and Empower2 chromatographic software).

[0087] The chromatographic conditions are as follows: the liquid chromatographic column adopts a general-purpose type, and the filler is selected from octadecyl bonded phase silica gel, such as Waters Atlantis T3C18 (4.6×250 mm, 5 μm). The flow rate is 1.0 mL / min; the injection volume is 10 μl; the column temperature is 32°C. All the other detection methods and conditions are with embodiment 1, and the liquid phase chromatogram result is as follows Figure 6 .

[0088] The minimum detection limit,...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention belongs to the field of analytical chemistry, and discloses a method for measuring multiple types of ATP (adenosine triphosphate) associated products in an aquatic product with a high performance liquid chromatography-diode array method. A high performance liquid chromatograph provided with a diode array detector is used for detection, and a to-be-detected sample is fed into the high performance liquid chromatograph for gradient elution after extracted; which ATP associated product each chromatographic peak belongs to is determined according to the retention time and the ultraviolet absorption spectrum of the ATP associated product, the corresponding ATP associated product chromatographic peak area is detected, and the content of the ATP associated products in a to-be-detected sample extracting solution is calculated according to an ATP associated product standard work curve or regression equation under the same chromatographic condition. According to the method, requirements for equipment and analysis conditions are simple, the content of multiple types of ATP associated products in the aquatic product can be measured simultaneously without adopting an ion-pairing agent, ten types of ATP associated products can be measured simultaneously, and the separating time doesn't exceed 25 minutes.

Description

technical field [0001] The invention belongs to the field of analytical chemistry and relates to a method for detecting multiple ATP-related products in aquatic products, in particular to a method for simultaneously measuring the contents of multiple ATP-related products in aquatic products by using high performance liquid chromatography-diode array method. Background technique [0002] ATP-related products are a class of compounds composed of purine bases, pyrimidine bases, nicotinamide, etc., and sugar phosphates, such as adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine ( Adenosine Monophosphate (AMP), Inosinic acid (IMP), Inosine (HxR), Hypoxanthine (Hx), Guanylic acid (GMP), Adenosine (Adenosine , AdR), adenine (Adenine, Ad), xanthine (Xanthine, Xt), etc. They are the main components of muscle nucleotides in aquatic animals, and are closely related to the flavor and freshness of aquatic raw materials. In terms of flavor, for example, AMP has the cha...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N30/02G01N30/06G01N30/74
Inventor 邱伟强陈舜胜谢晶曲映红施文正宋雪王丹妮王帅秦晓黄鑫晋颖颖
Owner SHANGHAI OCEAN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com