Ascorbic Acid

Applications:

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HPLC Separation of Two Vitamins: Different Polarity - Isocratic Methods

Primesep B and B2 columns separate Vitamins C (ascorbic acid) and B2 (riboflavin) with tunable selectivity. Peak order reversal is exhibited on these two columns with the same mobile phases due to their different polarity. The HPLC separation uses a mobile phase of water, acetonitrile (MeCN, ACN) and either formic or acetic acid (HOAc) with UV detection at 250 nm.

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Ascorbic Acid
Riboflavin (Vitamin B2)
Vitamin B2 (Riboflavin)
Vitamin C

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HPLC Separation of Organic Acids in HILIC Mode on Primesep N Column

Ascorbic, methylmalonic and succinic are weak organic acids. Retention of these three acids is achieved on Primesep N column in HILIC mode using acetonitrile/water and ammonium acetate. Compounds are monitored by ELSD. Method can be used for determination of ascorbic acid (Vitamin C), methylmalonic acid and succinic acid in various matrices. Other polar organic acids can be analyzed on this HILIC column.

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Ascorbic Acid
Methylmalonic Acid
Organic Acids
Succinic Acid

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HPLC Separation of Ascorbic and Dehydroascorbic Acids

Ascorbic acid is a vital vitamin for humans. It also serves as an antioxidant. Dehydroascorbic acid is an oxidation product of ascorbic acid. Both molecules are very polar, and cannot be retained by reverse-phase mechanism. Generic method for HPLC separation of ascorbic and dehydroascorbic acid was developed on a Primesep SB mixed-mode HPLC column. Both compounds are well separated and produce excellent peak shape. Method can be used for analysis of both compounds in biofluids. Proteins from biofluids will not retain due to repulsion effect of the stationary phase. Compounds can be monitored by common detection techniques like UV, ELSD, CAD, and LC/MS.

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Ascorbic Acid
Dehydroascorbic Acid

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HPLC Analysis of Active Drug in a Formulation

HPLC method for separation of active ingredients of drug/supplemental composition was developed on an Obelisc R trimodal HPLC column. Compounds are retained by combination of reversed-phase, cation-exchange and anion-exchange mechanisms. Compounds are well separated, and method can be used for quantitation of pyridoxine, ascorbic acid, niacinamide, pantothenic acid, caffeine and riboflavin in a mixture or as separate compounds in various complex mixtures. Various detection techniques can be applied for quantitation (ELSD, UV, LC/MS, Corona). This HPLC method can be adopted as general approach for analysis of active drug components in various formulations.

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Ascorbic Acid
Caffeine
Niacinamide
Pantothenic Acid
Pyridoxine
Riboflavin (Vitamin B2)

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HPLC Analysis of ACE Serum Active Ingredients

Mixed-mode HPLC columns allow for separation of compounds with drastically different properties. In this application, highly hydrophilic ascorbic acid is retained and separated from highly hydrophobic retinol and Vitamin E Acetate. Ascorbic acid is retained by anion-exchange mechanism and hydrophobic compounds are retained by reversed-phase mechanism. Compounds are monitored by UV detection. Mobile phase is compatible with LC/MS and method can be used to monitor basic and acidic compounds in bio-fluids (serum, blood, urine, saliva, etc.)

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Ascorbic Acid
Vitamin E Acetate

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HPLC Separation of Vitamin C, Vitamin Group B, and Related Impurities

Vitamin C (ascorbic acid) and Vitamins Group B are separated on Obelisc N mixed-mode column. Method can be used in quantitation and determination of polar vitamins in various formulations and dietary supplements. HPLC method can be based on UV, Evaporative Light Scattering Detection (ESLD), RI or MS detection. Effect of sample matrix can be eliminated by changing mobile phase conditions. Buffer concentration, buffer pH and amount of ACN will affect every vitamin differently due to difference in polar and ionic properties.

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3-Aminopyridine
4-Aminopyridine/Fampridine
Ascorbic Acid
Barbituric Acid
Isonicotinic Acid
Nicotinic Acid
Pyridoxine
Vitamin B2 (Riboflavin)
Vitamin B6 (Pyridoxine)

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HILIC Separation of Common Preservatives - Citric Acid, Ascorbic Acid and EDTA

Citric acid, ascorbic acid and EDTA are commonly used in food and pharmaceutical industry as preservatives. These compounds are very polar in nature. They are weak organic acids with limited UV activity. Retention and separation is achieved on HILIC mixed-mode Obelisc N column. All three compounds are retained by combination of strong HILIC and strong anion-exchange mechanisms. Separation can be monitored by ELSD, LC/MS, UV or Corona CAD. In contrast to other HILIC column, Obelisc N has two ionizable groups basic and acidic which provide ion-exchange interaction in addition to hydrophilic interaction. This allows to use less acetonitrile for HILIC separation.

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Ascorbic Acid
EDTA (Ethylenediaminetetraacetic Acid)
Vitamin C

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HPLC Separation of Organic Acids in HILIC and Anion-Exclusion Mode on Primesep S2 Column

Organic acids were separated on a HILIC/cation-exchange column in HILIC/anion-exclusion mode. This column can be used for analysis of polar compounds in HILIC mode. If compounds are ionizable, additional mode of interaction can be added (cation-exchange or anion-exclusion).

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1,5-Naphthalenedisulfonic Acid
3,5-Dihydroxybenzoic Acid
Ascorbic Acid
Organic Acids
p-Toluenesulfonic Acid (PTSA)

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HPLC Method for Analysis of Ascorbic Acid and Sodium Metabisulfite

Ascorbic is found naturally in citrus fruits and many vegetables. As a medication it is used to prevent or treat low levels of vitamin C, since it is that vitamin. Vitamin C is needed to maintain the health of skin, cartilage, teeth, bone, and blood vessels. Sodium metabisulfite is a white crystalline or powder solid. It has many uses, but some of it’s more prominent are: as the source of SO2 in wine, as a bleaching agent in the production of Coconut cream, and added to anaesthetic solutions to prevent oxidation to improve the shelf life of the solution. Primesep SB, a reverse-phase column, contains embedded basic ionizable groups and can retain Ascorbic acid and Sodium metabisulfite. The method is UV compatible and can be used as a general approach for analyzing similar compounds.

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Ascorbic Acid
Sodium metabisulfite

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HPLC Determination of Ascorbic Acid in Strawberry Juice

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FlipLC™ is an alternative method to avoid the interference of most of the contaminants by the use of an isolation column and a high pressure switching valve before the separation column. This method allows sample cleaning and analyte separation in one automated process. The isolation column and the separation column should have orthogonal retention characteristics to operate efficiently in this setup. Mixed-mode columns with reverse phase and ion-exchange characteristics were used in this analysis.

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Ascorbic Acid

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