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A blog on the latest applications, articles, & research on chromatography solutions in sample preparation, Ion Chromatography (IC),
High Pressure Liquid Chromatography (HPLC), Ion Chromatography-Mass Spectrometry (IC-MS), Gas Chromatography (GC),
Gas Chromatography-Mass Spectrometry (GC-MS and GC-MS/MS), and software (Chromeleon CDS, LIMS, and ProteinCenter).

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Chromatography Analysis for Vitamins (Part I)


chromatography analysis of Vitamin Waters 1This is the first of three blog posts on analytical solutions for vitamins in nutritional supplements and functional waters to ensure product quality and regulatory compliance (US FDA dietary supplement regulatory information) as well as to monitor vitamin intake. The market for both areas continues to grow rapidly around the globe. Even though the United States continues to be the largest market for nutritional and dietary supplements (Business Wire story on a comprehensive report), according to this story, the nutritional supplement market in India is expected to double by 2013. (In case you are interested, the story also provides an overview of the regulatory situation in India for nutritional supplements.)

As a part of our analytical solutions for food and beverage safety, our application labs have been paying close attention to nutritional supplements and have developed a large number of analytical methods. Here we feature two application notes on testing vitamins. 

Integrated, Efficient HPLC-UV Method for Water- and Fat-Soluble Vitamins in Nutritional Supplements

Typically, ensuring supplements contain the labeled amounts of vitamins can be time-consuming and difficult. Application Note 251, Determination of Water- and Fat-Soluble Vitamins in Nutritional Supplements by HPLC with UV Detection, (downloadable PDF) provides an integrated and efficient HPLC method using UV detection and Thermo Scientific Acclaim Reversed-Phase LC columns for determining water- and fat-soluble vitamins in nutritional supplements. Water-soluble vitamins, including seven B-group vitamins and Vitamin C, were well-resolved on the Acclaim Polar Advantage (PA) II column using a phosphate buffer. In this approach, the highly polar Vitamin C is well-retained without using an ion-pairing reagent. Fat-soluble vitamins, including Vitamins A, A acetate, E, D2, D3, K1, and β-carotene, were well resolved on the Acclaim 120 C18 column using an organic-solvent mobile phase. Method performance, including linearity, detection limits, reproducibility, and analyte recovery, was evaluated for both methods.

Determination of Vitamins in Functional Waters by HPLC-UV-PDA Detection

Functional beverages are vitamin-enhanced waters that have gained consumer popularity for convenience, perceived health benefits, and improved flavor over tap water. These beverages are typically enriched with A, B, B-complex, C, and E vitamins, with advertised benefits of increased energy from B vitamins and antioxidant benefits from A, C, and E vitamins. Industry forecasts predict increased sales of these beverages in the coming years. The US FDA requires these beverages to be labeled with nutritional content, thus creating the need for robust vitamin assays. Application 216, Determination of Water- and Fat-Soluble Vitamins in Functional Waters by HPLC with UV-PDA Detection, (downloadable PDF) discusses a method that provides a rapid means of analyzing a sample for both water- and fat-soluble vitamins in a single injection using a low flow rate that minimizes mobile phase preparation time and reduces waste.

In case you are interesting in testing for other compounds in nutritional supplements, visit our website for analytical applications for a range of dietary and herbal supplements.

Next blog post on measurement of fat-soluble vitamins and antioxidants, and separation of water and fat soluble vitamins in a single analysis, and separation of Vitamin K isomers!

Also, don't hesitate to comment below if you would like more details or found this information useful. We look forward to hearing from you!


Thanks for the info on the AP advantage. I need a clearification on the extraction of water soluble vitamins in water or alkaline medium.
Posted @ Sunday, April 14, 2013 2:51 PM by Kehinde Moyib
Hi Kehinde 
I will check with an expert and let you know. Thanks for your question. 
best regards 
Posted @ Monday, April 15, 2013 2:25 PM by Sonya Pelia
Hi Kehinde: 
Just heard back from the expert (he was traveling). 
The water soluble vitamins were not extracted in alkaline media. The vitamin B9 (folic acid) standard was prepared in 25 mM potassium hydrogen carbonate (alkaline) to make a concentrated standard. 
best regards 
Posted @ Monday, April 22, 2013 1:00 PM by Sonya Pelia
Dear Sonya, 
Do you have a similar application for quantify B-vitamin content in Fish meat?
Posted @ Wednesday, March 26, 2014 7:42 AM by Pat
Hi Patrick, 
My apologies for the delay in responding to your comment. It just appeared in my mailbox. (I have opened a tech support ticket to see why it took so long.) 
I have forwarded your question to our experts and expect to hear back shortly from them. 
best regards  
Posted @ Monday, May 19, 2014 11:17 AM by Sonya Pelia
Hi Patrick, 
We do not have an application for the analysis you mention. 
best regards 
Posted @ Monday, May 19, 2014 11:49 AM by Sonya Pelia
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