Ascorbic Acid

CAS Number 62624-30-0
Molecular Formula C6H8O6
Molecular Weight 176.124 g/mol
InChI Key CIWBSHSKHKDKBQ-UHFFFAOYSA-N
LogP -2.24
Synonyms
  • Ascorbic acid
  • (5S)-5-[(1R)-1,2-Dihydroxyethyl]-3,4-dihydroxyfuran-2(5H)-one (non-preferred name)
  • 62624-30-0
  • DL-Ascorbic acid
  • (5S)-5-[(1R)-1,2-Dihydroxyethyl]-3,4-dihydroxyfuran-2(5H)-one (non-preferred name)
  • EINECS 263-644-3

Applications:


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.



Application Analytes:

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

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.



Application Analytes:

Ascorbic Acid
Methylmalonic Acid
Organic Acids
Succinic Acid

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.



Application Analytes:

Ascorbic Acid
Dehydroascorbic Acid

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.



Application Analytes:

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

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.)



Application Analytes:

Ascorbic Acid
Vitamin E Acetate

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.



Application Analytes:

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

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.



Application Analytes:

Ascorbic Acid
EDTA (Ethylenediaminetetraacetic Acid)
Vitamin C

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).

Condition

Column Primesep S2, 4.6x150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O - 85/15%
Buffer AmAc pH 5.0 15 mM
Flow Rate 1.0 ml/min
Detection ELSD, 50C UV 250 nm
 

Description

Class of Compounds Nucleosides,  Hydrophilic, Ionizable
Analyzing Compounds Toluenesulfonic acid, Naphthalenedisulfonic acid, 3.5 DHBA, Ascorbic acid,
 

Application Analytes:

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

HPLC Method for Analysis of Ascorbic Acid and Sodium Metabisulfite

*The metabisulfite ion (S2O52−) is hydrolyzed to bisulfite (HSO3) in water. 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 anesthetic solutions to prevent oxidation to improve the shelf life of the solution. 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. Primesep SB, a reverse-phase column, contains embedded basic ionizable groups and can retain Ascorbic acid and Sodium bisulfite. The method is UV compatible and can be used as a general approach for analyzing similar compounds.

 

Condition

Column Primesep SB, 4.6x150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Buffer H3PO4
Flow Rate 1.0 ml/min
Detection UV, 270 nm
 

Description

Class of Compounds Acid, Hydrophilic, Ionizable, Disinfectant, Antioxidant, Preservative agent.
Analyzing Compounds Ascorbic Acid, Sodium Metabisulfite
 

Application Analytes:

Ascorbic Acid
Sodium bisulfite
Sodium metabisulfite

HPLC Determination of Ascorbic Acid in Strawberry Juice

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.

Application Analytes:

Ascorbic Acid