Succinic Acid

Separation type: Bridge Ion Separation Technology, or BIST™ by SIELC Technologies

High Performance Liquid Chromatography (HPLC) Method for Analyses of Succinic Acid

Methanesulfonic acid is a popular non-volatile catalyst used in organic reactions due to it being a strong acid. Other sulfonic acids, like 1-Pentanesulfonic acid, 1-Heptanesulfonic acid, 1-Decanesulfonic acid, and 1=Dodecanesulfonic acid are typically used in ion chromatography and for organic syntheses. Using SIELC’s newly introduced BIST™ method, a mixture of these Sulfonic acids can be separated on a negatively-charged, cation-exchange BIST™ A+ column, contrary to conventional chromatographic wisdom. There are two keys to this retention method: 1) a multi-charged, positive buffer, such as N,N,N’,N’-Tetramethyl-1,3-propanediamine (TMDAP), which acts as a bridge, linking the negatively-charged anion analytes to the negatively-charged column surface and 2) a mobile phase consisting mostly of organic solvent (such as MeCN) to minimize the formation of a solvation layer around the charged analytes. Other positively-charged buffers that can generate BIST™ include Calcium acetate and Magnesium acetate. Using this new and unique analysis method, these Sulfonic acids can be separated, retained, and detected through ELSD. This method is also compatible with Mass Spectrometry (LC-MS) and CAD.

Condition

Description

Separation type: Liquid Chromatography Mixed-mode

Succinic acid is a key intermediary (in the form of its anion, succinate) in the electron transport chain, a process that is the key power source for our cells. It also has a few limited functions outside of the mitochondria. Glutaric acid is a common plasticizer precursor and often used in the production of polyester polyols and polyamides. Interestingly, it is significantly more water soluble than either Succinic or Adipic acid. Adipic acid, also known as hexanedioic acid, is a key precursor to nylon, and more than 2.5 billion kilograms are produced every year. Methylmalonic acid is a key intermediary in the anaplerotic reactions of the Krebs cycle. These Dibasic acids can be separated, retained, and analyzed on a mixed-mode Newcrom B column with a mobile phase consisting of water, Acetonitrile (MeCN), and Sulfuric acid (FA). This analytical method can be UV detected at 200 nm with high resolution and peak symmetry.

High Performance Liquid Chromatography (HPLC) Method for Analysis of Adipic Acid, Succinic Acid, Glutaric Acid, Methylmalonic acid

Separation type: Liquid Chromatography HILIC

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High Performance Liquid Chromatography (HPLC) Method for Analysis of Succinic Anhydride and Succinic Acid

Succinic acid is a key intermediary (in the form of its anion, succinate) in the electron transport chain, a process that is the key power source for our cells. It also has a few limited functions outside of the mitochondria. Succinic anhydride, its acid anhyrdie, is a colorless, crystalline compound used often in the production of polymer resins, often of the alkyd variety. It also has the ability to increase the solubility of various proteins and has other benefits for the pharmaceutical, flavor, and fragrance industries.
Succinic Anhydride and Succinic Acid can be detected in the low UV regime. Using a SHARC 1 hydrogen-bond column and a mobile phase consisting of only acetonitrile (and without a buffer), these two compounds can be separated, detected, and analyzed. This analysis method can be UV detected at 210 nm.

*Before running, wash the column using 0.5 % phosphoric acid in water and run this MP for a couple of hours. Then switch to pure MeCN.

Separation type: Liquid Chromatography HILIC

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High Performance Liquid Chromatography (HPLC) Method for Analysis of Succinic Anhydride and Succinic Acid

Succinic acid is a key intermediary (in the form of its anion, succinate) in the electron transport chain, a process that is the key power source for our cells. It also has a few limited functions outside of the mitochondria. Succinic anhydride, its acid anhydride, is a colorless, crystalline compound used often in the production of polymer resins, often of the alkyd variety. It also has the ability to increase the solubility of various proteins and has other benefits for the pharmaceutical, flavor, and fragrance industries.
These two related compounds can be detected in the low UV regime. Using a Primsep S2 HILIC column, which uses hydrogen-bonding as a separation mechanism, and a mobile phase consisting of water and an acetonitrile (ACN) gradient with no buffer, both of these compounds can be separated and retained. This analysis method can be UV detected at 225 nm.

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The citric acid cycle (tricarboxylic acid cycle, Krebs cycle) is a key process in the metabolic pathway by which all aerobic organisms generate energy. Compounds generated during TCA are hydrophilic, acidic compounds. Some of the hydrophilic acids are very strong while others are relatively weaker. All compounds have very low hydrophobicity and do not retain by the reversed-phase mechanism on the C18 or C8 HPLC columns. Citric acid and related products were separated in reversed-phase, anion-exchange mode on the Primesep D mixed-mode column. All hydrophilic, acidic compounds are well separated and demonstrate good shape. This robust method can be used for analysis of components of the TCA cycle and other highly hydrophilic compounds. This method is fully compatible with LC/MS and prep chromatography.

This is an alternative approach for the separation of the TCA cycle intermediates that was developed using novel hydrogen-bonding HPLC column. The organic nature of the mobile phase helps obtain a highly sensitive LC/MS compatible method.

Primesep D mixed-mode column separates organic acids such as succinic acid, malic acid, MPS and butanesulfonate by a mixture of anion exchange and reversed phase mechanisms. Retention times can be changed by adjusting the percentage of acetonitrile in the mobile phase. This can not be done by traditional ion-exchange and ion-exclusion chromatography. The HPLC separation uses a mobile phase of water, acetonitrile (MeCN, ACN) and ammonium formate as a buffer, making the method MS-compatible. Can also use UV detection at 250 nm.

Organic and inorganic acids can be retained and separated on mixed-mode columns based on weak reversed-phase and weak/medium anion-exchange mechanisms. Amount of ACN, buffer concentration and buffer pH will affect retention time of organic and inorganic acids. Acids can be monitored by low UV, ELSD or LC/MS. Presence of ions is required to facilitate ion-exchange mechanism. Method can be used as a general approach for analysis of acidic hydrophilic and acidic hydrophobic compounds. Carboxylic acids along with inorganic acid can be retained and separated without ion-pairing reagent.

Organic and inorganic acids can be separated by reversed-phase anion-exchange Primesep D column. Succinic and methylmalonic acids were separated based on their hydrophobic and ionic properties. Method can be used for analysis of other acids by HPLC.

In mixed-mode HILIC chromatography, selectivity of separation can be adjusted by amount of acetonitrile, amount of buffer and buffer pH. Buffer concentration and pH will affect retention of ionizable compounds to a different degree. Retention of neutral compounds can be adjusted by the amount of acetonitrile. Carboxylic acid, three amino acids and two sugars are separated by combination of HILIC and ion-exchange mechanisms. Compounds can be monitored by ELSD, Corona (CAD), LC/MS or low UV. UV-transparent mobile phase /buffer is required for UV monitoring of this mixed-mode separation. This HPLC method can be adopted as general approach for analysis of sugars, amino acids and carboxylic acids.

The majority of drugs in the pharmaceutical industry are administered in salt form. The presence of two counter-ions very often necessitates the use of two methods. The nature of these counterparts in drugs can be an inorganic cation and organic acid, inorganic anion and organic base, and organic cation and organic anion. Furthermore, the properties of the molecules will result in a differing stoichiometry. The task of simultaneous quantitation of counter-ions can be achieved by using mixed-mode columns. The general approach for analysis is based on properties of corresponding counter-ions. Hydrophobic basic drugs, like dextromethorphan, verapamil, trimipramine, and corresponding acidic counter-ions (chloride, chlorate, bromide, bromate, perchlorate, maleate, fumarate,tartrate, succinate, phosphate, citrate, benzosulfonate, toleuensulfonate) can be separated and quantitated in the same run on reversed-phase anion-exchange column. Basic hydrophobic drugs are retained by the reversed-phase mechanism, and counter-ions are retained by the reversed-phase and anion-exchange mechanism. Some polar counter-ions are retained only by the anion-exchange mechanism. Retention time and selectivity of HPLC separation of drugs and counter-ions can be achieved by changing the amount of acetonitrile and the amount of ions in the mobile phase. The detection technique depends on the properties of the counter-ions. In case of low or no UV activity, ELSD can be employed if the counter-ion forms a non-volatile salt with the mobile phase additive (ammonium formate). This HPLC method can be used for simultaneous quantitation of other basic drugs and counter-ions. The presence of two mechanisms of retention allows control over retention times of drug and counter-ion independently, and even allows a change of order of elution when necessary.

Hydrophilic acids are separated on Obelisc N mixed-mode HILIC column. Seven carboxylic acids are separated based on their polarity and pKa values. Changes in ionization states of acids and stationary phase can be used to control elution order of organic and inorganic acids.

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.

Succinic acid is a dicarboxylic acid that plays important role in citric acid cycle. Succinic acid is byproduct of sugar fermentation, and is present in wines and beer. In one of the forms it exists as a salt with sodium. Both sodium and succinic acid can be retained and separated on Primesep 100 column. Sodium is retained by cation-exchange mechanism, while succinic acid is separated by reverse phase mechanism. Method can be used for HPLC analysis of metal ions (sodium, potassium, lithium, calcium, etc) and carboxylic acids (hydrophobic carboxylic mono and diacids) in one chromatographic run. ELSD is detection of choice for non-UV active non-volatile compounds.

Primesep 200 retains and separates the organic diacids (malic, succinic, fumaric, and maleic) by a combination hydrophobic, reversed-phase interactions and ion exclusion. The separation uses a mobile phase of water, acetonitrile (MeCN, ACN) and trifluoracetic acid (TFA) with UV detection at 210 nm.

Primesep 100 separates a mixture of dicarboxylic acids in ion-exclusion mode with a mobile phase of water, acetonitrile (MeCN, ACN), and sulfuric acid (H2SO4) with UV detection at 210 nm. Baseline resolution of fumaric, maleic, malic, and succinic acids is obtained in less than 8 minutes. The separation combines ion-exclusion and reversed-phase mechanisms in one method.