We use cookies to understand how you use our site and to improve your experience. This includes personalizing content and advertising. To learn more, click here. By continuing to use our site, you accept our use of cookies. Cookie Policy.

Features Partner Sites Information LinkXpress hp
Sign In
Advertise with Us
PURITAN MEDICAL

THERMO FISHER SCIENTIFIC

Thermo Fisher Scientific provides analytical instruments, lab equipment, specialty diagnostics, reagents and integrat... read more Featured Products: More products

Download Mobile App




HPLC Method Developed for Vitamin B6 Quantitation

By LabMedica International staff writers
Posted on 21 Apr 2020
Symptoms of vitamin B6 deficiency can include dermatitis, glossitis, depression, confusion, convulsions, and anemia. More...
Inborn errors affecting vitamin B6 metabolism have been associated with vitamin B6-dependent epilepsy.

While vitamin B6 status is primarily assessed by measuring the biologically active form of the vitamin, pyridoxal 5-phosphate (PLP), concurrent measurement of the final metabolite 4-pyridoxic acid (PA) can provide additional information regarding supplement intake and hypophosphatasia.

Laboratory Scientists at the Cleveland Clinic (Cleveland, OH, USA) developed a one-step reverse phase High-Performance Liquid Chromatography (HPLC) method with fluorescence detection by evaluating different derivatization conditions, the use of an internal standard and different calibration strategies. The assay analytical performance was evaluated. A Thermal Vanquish UHPLC system equipped with fluorescence detector (Thermo Fisher Scientific, Waltham, MA, USA) was used in the analysis.

The team reported that pre-column derivatization with semicarbazide showed the best overall performance in terms of signal to noise ratio, retention time and peak shape when compared to pre- or post-column derivatization with chlorite, pre-column or in-mobile phase derivatization using sodium bisulfite. The final method provided an analytical measurement range from 7.7-300 nmol/L for PLP and 3.7–300 nmol/L for PA, total imprecision <15% and <5% for PLP and PA respectively. Calibration against the National Institute of Standards and Technology (NIST) standard produced measured values within 3% of NIST assigned PLP values. The use of 4-deoxypyridoxine as internal standard did not improve precision or accuracy when compared to calibration using 5-level external standards.

The authors concluded that they had developed a simple one-step HPLC method for evaluation of vitamin B6 status by measuring the bioactive PLP and final metabolite PA simultaneously. The assay procedure was simplified into a one-step approach by combining the protein precipitation and derivatization procedure without compromising analytical performance. The method is fast, precise and sensitive enough to detect vitamin B6 concentrations below 2.5th percentile among the general population in the USA.



Gold Member
Antipsychotic TDM Assays
Saladax Antipsychotic Assays
Serological Pipet Controller
PIPETBOY GENIUS
New
Gold Member
Quality Control Material
iPLEX Pro Exome QC Panel
New
Staining Management Software
DakoLink
Read the full article by registering today, it's FREE! It's Free!
Register now for FREE to LabMedica.com and get access to news and events that shape the world of Clinical Laboratory Medicine.
  • Free digital version edition of LabMedica International sent by email on regular basis
  • Free print version of LabMedica International magazine (available only outside USA and Canada).
  • Free and unlimited access to back issues of LabMedica International in digital format
  • Free LabMedica International Newsletter sent every week containing the latest news
  • Free breaking news sent via email
  • Free access to Events Calendar
  • Free access to LinkXpress new product services
  • REGISTRATION IS FREE AND EASY!
Click here to Register








DIASOURCE (A Biovendor Company)

Channels

Hematology

view channel
Image: CitoCBC is the world first cartridge-based CBC to be granted CLIA Waived status by FDA (Photo courtesy of CytoChip)

Disposable Cartridge-Based Test Delivers Rapid and Accurate CBC Results

Complete Blood Count (CBC) is one of the most commonly ordered lab tests, crucial for diagnosing diseases, monitoring therapies, and conducting routine health screenings. However, more than 90% of physician... Read more

Immunology

view channel
Image: A simple blood test could replace surgical biopsies for early detecion of heart transplant rejection (Photo courtesy of Shutterstock)

Blood Test Detects Organ Rejection in Heart Transplant Patients

Following a heart transplant, patients are required to undergo surgical biopsies so that physicians can assess the possibility of organ rejection. Rejection happens when the recipient’s immune system identifies... Read more

Pathology

view channel
These images illustrate how precision oncology Organ Chips recapitulate individual patients’ responses to chemotherapy (Photo courtesy of Wyss Institute at Harvard University)

Cancer Chip Accurately Predicts Patient-Specific Chemotherapy Response

Esophageal adenocarcinoma (EAC), one of the two primary types of esophageal cancer, ranks as the sixth leading cause of cancer-related deaths worldwide and currently lacks effective targeted therapies.... Read more
Copyright © 2000-2025 Globetech Media. All rights reserved.