Skip to content

JASCO JASCO

  • News
  • Events
  • E-Store
  • My Account
  • Contact Us
  • Worldwide
Search
Click to view menu
MENUMENU
  • Products
    • Chromatography
      • HPLC
      • RHPLC
      • UHPLC
      • LC-MS
      • Preparative LC
      • Analytical SFC
      • Semi-Preparative SFC
      • Hybrid SFC
      • Fuel Analysis by SFC-FID
      • Preparative SFC
      • Supercritical Fluid Extraction
      • Chromatography Software
    • Molecular Spectroscopy
      • Circular Dichroism
      • High-Throughput CD
      • Vibrational CD
      • Circularly Polarized Luminescence (CPL)
      • Polarimeters
      • FTIR Spectrometers
      • FTIR Microscopy
      • FTIR Portable
      • Raman Microscopy
      • Palmtop Raman Spectrometer
      • Probe Raman
      • UV-Visible/NIR Spectrophotometers
      • UV-Visible/NIR Microscopy
      • Fluorescence Spectrophotometers
      • Film Thickness
      • Spectra Manager™ Suite
    • Refurbished
      • Refurbished HPLC Systems
      • HPLC Switching Valves
      • FTIR Accessories
  • Service
    • Service and Support Plans
    • Service Request Form
  • Applications
  • KnowledgeBase
  • Learning Center
    • Best Practice
      • Circular Dichroism Tips & Tricks for Biological Samples
      • CD Scale Calibration with ACS
      • Fluorescence Tips & Tricks
      • Raman Spectroscopy Tips & Tricks
    • Training Videos
      • ChromNAV
      • SF-NAV
      • Circular Dichroism
      • UV-Visible/NIR
      • Fluorescence
    • Training Seminars
      • Training Registration Form
    • Webinars
    • eBooks
    • Theory
      • Theory of Molecular Spectroscopy
      • Chromatography
  • About Us
    • President’s Message
    • Contact
    • History
    • Careers
  • News
  • Events
  • Worldwide
  • Shop
  • My Account
  • Contact Us

Home / Applications / Speed Separation of ATP and its Degradation Products By X-LC

  • Industry

  • Technique

Speed Separation of ATP and its Degradation Products By X-LC

By Satoko Suzuki

PDF IconDownload This Application

January 5, 2024

Introduction

ATP (adenosine triphosphate) in fish meat decomposes in a following process:

ATP -> ADP (adenosine diphosphate) -> AMP (adenosine monophosphate) -> Inosinic acid -> Inosine -> Hypoxanthine

A parameter “K value” indicates freshness of fish meat and is defined as:

K value(%) = [(Inosine + Hypoxanthine) / (ATP + ADP + AMP + Inosine + Hypoxanthine)] x 100

When K value is below 20 %, fish meat can be used for Sashimi (raw use) and 20% through 60% for cooked and processed use. We applied X-LC to an analysis of K value and evaluated its performance in comparison with results of conventional HPLC.

Experimental

Jasco X-LC system for ATP and degradation products separation

The chromatography system utilized in this experiment consists of JASCO X-LC series: two of 3185PU solvent delivery pump, 3080DG degasser, 3180MX high pressure mixing device, 3067CO column oven, 3070UV UV/Vis detector, 3159AS autosampler, and ChromNAV chromatography data system. Separation column used was a X-PressPak AQ-C18-W (3.0 mm ID x 50 mmL, 2µm). K value was obtained by a program calculation function in ChromNAV.

Pretreatment of fish meat was conducted in the following procedure:

  1. Add 0.4 M perchloric acid solution (20 mL) to fish meat (2.5 g) to homogenize.
  2. Centrifuge it at 3500 rpm for 10 min.
  3. Add 2 M potassium carbonate solution (1 mL) to the supernatant (5 mL) and centrifuge it at 3500 rpm for 5 min.
  4. Filter supernatant obtained at (3) with 0.45 µm membrane filter.

Keywords

420008XRE

Results

Figure 1 shows an X-LC chromatogram of standard mixture. X-LC succeeded in providing an analysis time eight times shorter than conventional HPLC. Figure 2 shows an X-LC chromatogram of tuna preserved for two days. As shown in this figure, six kinds of components provide a good separation without interfering with unknown components. K value of this sample was 8%. Figure 3 shows relationship between preservation days and K values of tuna and sea bream.

Figure 1. X-LC chromatogram of a standard mixture
Peaks: 1=adenosine triphosphate (ATP), 2=adenosine diphosphaete (ADP), 3=inosinic acid (IMP), 4=hypoxanthine (Hypo),
5=adenosine monophosphate(AMP), 6=inosine (Ino) Chromatographic conditions:
Column=X-Presspak AQ-C18-W (3.0 mm ID x 50 mmL, 2.0 µm) Mobile phase: A=100mM phosphate buffer (pH 4.2),
B=100 mM phosphate buffer (pH 4.2)/ acetonitrile (50/50)
Gradient profile: 0min, A/B (100/0); 2.2 min, A/B (50/50); 7 min, A/B (50/50); 8.2 min, A/B (0/100); 8.3 min, A/B(100/0)
Flow rate: 0.6 mL/min Column temperature: 30°C Detection wavelength: 260 nm Injection volume: 1 µL (1nmol each)
Figure 2. X-LC chromatogram of tuna sample
The pretreatment is describe in Experimental. The other conditions are the same as in Figure 1 caption.
Figure 3. Relationship between preservation days and K values.
This document has been prepared based on information available at the time of publication and is subject to revision without notice. Although the contents are checked with the utmost care, we do not guarantee their accuracy or completeness. JASCO Corporation assumes no responsibility or liability for any loss or damage incurred as a result of the use of any information contained in this document. Copyright and other intellectual property rights in this document remain the property of JASCO Corporation. Please do not attempt to copy, modify, redistribute, or sell etc. in whole or in part without prior written permission.

Featured Products:

  • LC-4000 Series UHPLC

  • Tandem LC-MS

  • Single Quadrupole LC-MS

About the Author

Satoko Suzuki is a senior manager of the molecular spectroscopy applications laboratory at the JASCO facility in Hachioji Japan.

JASCO Application Note

Speed Separation of ATP and its Degradation Products By X-LC

Introduction

ATP (adenosine triphosphate) in fish meat decomposes in a following process:

ATP -> ADP (adenosine diphosphate) -> AMP (adenosine monophosphate) -> Inosinic acid -> Inosine -> Hypoxanthine

A parameter “K value” indicates freshness of fish meat and is defined as:

K value(%) = [(Inosine + Hypoxanthine) / (ATP + ADP + AMP + Inosine + Hypoxanthine)] x 100

When K value is below 20 %, fish meat can be used for Sashimi (raw use) and 20% through 60% for cooked and processed use. We applied X-LC to an analysis of K value and evaluated its performance in comparison with results of conventional HPLC.

Experimental

Jasco X-LC system for ATP and degradation products separation

The chromatography system utilized in this experiment consists of JASCO X-LC series: two of 3185PU solvent delivery pump, 3080DG degasser, 3180MX high pressure mixing device, 3067CO column oven, 3070UV UV/Vis detector, 3159AS autosampler, and ChromNAV chromatography data system. Separation column used was a X-PressPak AQ-C18-W (3.0 mm ID x 50 mmL, 2µm). K value was obtained by a program calculation function in ChromNAV.

Pretreatment of fish meat was conducted in the following procedure:

  1. Add 0.4 M perchloric acid solution (20 mL) to fish meat (2.5 g) to homogenize.
  2. Centrifuge it at 3500 rpm for 10 min.
  3. Add 2 M potassium carbonate solution (1 mL) to the supernatant (5 mL) and centrifuge it at 3500 rpm for 5 min.
  4. Filter supernatant obtained at (3) with 0.45 µm membrane filter.

Results

Figure 1 shows an X-LC chromatogram of standard mixture. X-LC succeeded in providing an analysis time eight times shorter than conventional HPLC. Figure 2 shows an X-LC chromatogram of tuna preserved for two days. As shown in this figure, six kinds of components provide a good separation without interfering with unknown components. K value of this sample was 8%. Figure 3 shows relationship between preservation days and K values of tuna and sea bream.

Figure 1. X-LC chromatogram of a standard mixture
Peaks: 1=adenosine triphosphate (ATP), 2=adenosine diphosphaete (ADP), 3=inosinic acid (IMP), 4=hypoxanthine (Hypo),
5=adenosine monophosphate(AMP), 6=inosine (Ino) Chromatographic conditions:
Column=X-Presspak AQ-C18-W (3.0 mm ID x 50 mmL, 2.0 µm) Mobile phase: A=100mM phosphate buffer (pH 4.2),
B=100 mM phosphate buffer (pH 4.2)/ acetonitrile (50/50)
Gradient profile: 0min, A/B (100/0); 2.2 min, A/B (50/50); 7 min, A/B (50/50); 8.2 min, A/B (0/100); 8.3 min, A/B(100/0)
Flow rate: 0.6 mL/min Column temperature: 30°C Detection wavelength: 260 nm Injection volume: 1 µL (1nmol each)
Figure 2. X-LC chromatogram of tuna sample
The pretreatment is describe in Experimental. The other conditions are the same as in Figure 1 caption.
Figure 3. Relationship between preservation days and K values.

Keywords

420008XRE

This document has been prepared based on information available at the time of publication and is subject to revision without notice. Although the contents are checked with the utmost care, we do not guarantee their accuracy or completeness. JASCO Corporation assumes no responsibility or liability for any loss or damage incurred as a result of the use of any information contained in this document. Copyright and other intellectual property rights in this document remain the property of JASCO Corporation. Please do not attempt to copy, modify, redistribute, or sell etc. in whole or in part without prior written permission.
28600 Mary’s Court, Easton, MD 21601 USA • (800) 333-5272 • Fax: (410) 822-7526 • jascoinc.com/applications

Close

Designed in Tokyo. TRUSTED globally.

View our support plans

Connect with JASCO

  • Facebook
  • Twitter
  • LinkedIn
  • JASCO Sales
  • 800-333-5272

Receive the latest promotions and special offers

  • This field is for validation purposes and should be left unchanged.
  • Careers
  • Press Kit
  • JASCO Privacy Policy
  • Sitemap
  • Environmental Policy

© , JASCO. All Rights Reserved.