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 / High Speed Separation of Isoflavones in Soybean Milk Using UHPLC

  • Industry

  • Technique

High Speed Separation of Isoflavones in Soybean Milk Using UHPLC

By Kohei Tamura

PDF IconDownload This Application

August 18, 2022

Introduction

Isoflavones are polyphenolic compounds produced in Fabaceae such as soybeans. They act as phytoestrogens in mammals. They are also strong antioxidants and considered useful in treating cancer. Common isoflavones include daizin, genistin, glycitin, daidzein, genistein, and glycitein.

We have demonstrated, using extreme high-pressure liquid chromatography ( X-LC ) the analysis of isoflavones. The results were obtained using an X-PressPak C18S (2.1 mm I.D. x 50 mmL., 2 µm) column combined with a high-pressure gradient method described below. Total analysis time was approximately 4 minutes.

Jasco XLC-3000

Experimental

The system used for the measurement consisted of two 3185PU pumps, 3080DG degasser, 3180MX mixing unit, 3067CO column oven, 3070UV UV/Vis detector 3059AS autosampler and a chromatography data system.

Soymilk (2 mL) is mixed with methanol (2 mL), incubated for 90 minutes at 60°C, mixed with hexane (1 mL), and centrifuged. The supernatant is filtered with 0.45 µm membrane filter. A portion of 1 µL is injected.

Results

Figure 1 shows the chromatogram of a standard mixture of isoflavones. Each component is sufficiently separated; the analysis time is 4 minutes and the time from injection to injection is 7.5 minutes.

Figure 2 shows the chromatogram of soymilk. Each component is well separated from unknown peaks.

Figure 1.X-LC chromatogram of a standard mixture of isoflavone.
Conditions: column = X-PressPak C18S (2.1 mm I.D. x 50 mmL.), column temperature = 40°C, Mobile phase: A = acetonitrile/water/acetic acid (10/90/0.1), B = acetonitrile/water/acetic acid (40/60/0.1), 0 min = A/B(100/0) — 5 min = A/B(0/100) — 5.5 min = A/B(0/100) — 5.55 min = A/B(100/0), flow rate = 0.6 mL/min, detection wavelength = 254 nm, injection volume = 1mL
Figure 2Chromatogram of soybean milk.
The conditions are the same as in figure 1. Pretreatment: 1. Add methanol (2 mL) to soybean milk (2 mL), 2. Stand at 60°C for 1.5 hours, 3. Add Hexane (1 mL) and mix, 4. The hexane layer is filtered.
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:

  • Single Quadrupole LC-MS

  • Column Ovens

  • UV-Visible Detectors

About the Author

Mr. Tamura is a member of the applications team with a specialty in vibrational spectroscopy and particularly Raman and IR Imaging. He is also one of our experts in gas analysis by FTIR.

JASCO Application Note

High Speed Separation of Isoflavones in Soybean Milk Using UHPLC

Introduction

Isoflavones are polyphenolic compounds produced in Fabaceae such as soybeans. They act as phytoestrogens in mammals. They are also strong antioxidants and considered useful in treating cancer. Common isoflavones include daizin, genistin, glycitin, daidzein, genistein, and glycitein.

We have demonstrated, using extreme high-pressure liquid chromatography ( X-LC ) the analysis of isoflavones. The results were obtained using an X-PressPak C18S (2.1 mm I.D. x 50 mmL., 2 µm) column combined with a high-pressure gradient method described below. Total analysis time was approximately 4 minutes.

Jasco XLC-3000

Experimental

The system used for the measurement consisted of two 3185PU pumps, 3080DG degasser, 3180MX mixing unit, 3067CO column oven, 3070UV UV/Vis detector 3059AS autosampler and a chromatography data system.

Soymilk (2 mL) is mixed with methanol (2 mL), incubated for 90 minutes at 60°C, mixed with hexane (1 mL), and centrifuged. The supernatant is filtered with 0.45 µm membrane filter. A portion of 1 µL is injected.

Results

Figure 1 shows the chromatogram of a standard mixture of isoflavones. Each component is sufficiently separated; the analysis time is 4 minutes and the time from injection to injection is 7.5 minutes.

Figure 2 shows the chromatogram of soymilk. Each component is well separated from unknown peaks.

Figure 1.X-LC chromatogram of a standard mixture of isoflavone.
Conditions: column = X-PressPak C18S (2.1 mm I.D. x 50 mmL.), column temperature = 40°C, Mobile phase: A = acetonitrile/water/acetic acid (10/90/0.1), B = acetonitrile/water/acetic acid (40/60/0.1), 0 min = A/B(100/0) — 5 min = A/B(0/100) — 5.5 min = A/B(0/100) — 5.55 min = A/B(100/0), flow rate = 0.6 mL/min, detection wavelength = 254 nm, injection volume = 1mL
Figure 2Chromatogram of soybean milk.
The conditions are the same as in figure 1. Pretreatment: 1. Add methanol (2 mL) to soybean milk (2 mL), 2. Stand at 60°C for 1.5 hours, 3. Add Hexane (1 mL) and mix, 4. The hexane layer is filtered.
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.