Extraction pantothenic acid

For most assays, the incorporated pantothenic acid has to be Hberated en2ymatically. Usually, a combination of pantotheinase and alkaline phosphatase is used to hberate the bound pantothenic acid. The official method for pantothenic acid of the Association of Official Analytical Chemists (AOAC) is the microbiological assay that uses U. Plantarium (A.TCC 8014) as the test organism (71). Samples are extracted at 121°C at pH 5.6—5.7, proteins are precipitated at pH 4.5, and the resulting clear extracts are adjusted to pH 6.8 prior to assay. This procedure is only suitable to determine calcium pantothenate or other free forms of pantothenic acid. 

Pantothenic acid is found in extracts from nearly all plants, bacteria, and animals, and the name derives from the Greek pantos, meaning everywhere. It is required in the diet of all vertebrates, but some microorganisms produce it in the rumens of animals such as cattle and sheep. This vitamin is widely distributed in foods common to the human diet, and deficiencies are only observed in cases of severe malnutrition. The eminent German-born biochemist Fritz Lipmann was the first to show that a coenzyme was required to facilitate biological acetylation reactions. (The A in... 

Pantothenic acid occurs in foods both in the free form and bonded to coenzyme (CoA) or acyl carrier protein (ACP) therefore hydrolysis is needed to extract it totally. Since it is degraded by acid and alkaline hydrolysis, only an enzymatic digestion can be applied. Enzyme hydrolysis with papain, diastase, clarase, takadiastase, intestinal phosphatase, pigeon liver pantetheinase, or combination of them has been used. 

More recently [635], a unique extraction step in supplemented foods, by using hot water and a precipitation solution, following by HPLC-ELD/UV analysis has been performed for the simultaneous determination of pyridoxine, thiamine, riboflavin, niacin, pantothenic acid, folic acid, cyanoco-balamin, and ascorbic acid. The mobile phase consisting of phosphate buffer and methanol has been modified in order to perform ion-liquid chromatography by adding l-octanesulfonic acid sodium salt. Furthermore, triethylamine has been also added to improve peak symmetry. 

Enzyme hydrolysis, with papain, diastase, clarase, takadiastase, intestinal phosphatase, or combinations thereof is most commonly used to release pantothenate from food proteins (186). A cold perchloric acid extraction was used to release pantothenic acid from tissue samples (187). Food spoilage prior to analysis may lead to inflated pantothenic acid levels (19). 

Some itamirLS are water soluble, while others are fat soluble. This classification is valuable as it indicates whether the vitamin is likely to be absorbed similarly to lipids or like other water-soluble nutrients. The fat-soluble vitamins are A, D, E, and K. The water-soluble vitamins arc ascorbic acid, biotin, folate, niacin, pantothenic acid, riboflavin, thiamin, vitamin B i, and vitamin B 2. The classification is also valuable, as it helps chemists decide on the best way to extract and analyze a particular vitamin in foods and biological tissues. Aside from having some bearing on the path ways of absorption and distribution throughout the body, the question of whether a particular vitamin is fat soluble or water soluble has little or no relevance to its function in the body. 

A rapid, simple, and specific TLC method has been developed for the estimation of panthenol and pantothenic acid in pharmaceutical preparations containing other vitamins, amino acids, symps, enzymes, etc. " The vitamin was extracted with ethanol (from tablets and capsules) or benzyl alcohol (from liquid oral preparations) and isolated from other ingredients by TLC on silica gel 60 plates with 2-propanol-water (85 15 vol/vol) as a solvent. p-Alanine (panthothenate) or p-alanol (panthenol) was liberated by heating for 20 min at 160°C. The liberated amines were visualized with the ninhydrin reaction and estimated by spectrodensitometry at 490 nm. Recoveries for panthenol and pantothenic acid were 99.8 2.25% and 100.2 1.7%, respectively. 

It is well known that in the oxidation of pyruvate an active 2-carbon intermediate, known as active acetate, is formed (Gurin and Crandall, 1948 Bloch, 1948 Lehninger, 1950). A suggestion that pantothenic acid is involved in the conversion of acetate to citrate in yeast came from Novelli and Lipmann s (1950) experiments showing a correlation between the ability of the yeast to oxidize acetate or ethanol and the coenzyme A content of the cells. The formation of citrate from acetate and oxaloacetate in cell-free extracts from pigeon liver or yeast was also shown to require coenzyme A (Novelli and Lipmann, 1950 Stern and Ochoa, 1949, 1951). Further study of these enzyme systems led to the isolation of a crystalline condensing enzyme from pig heart (Ochoa et al., 1951) and to a formulation of the mechanism of the enzymatic synthesis of citric acid from... 

However, if the deficient animals received adrenocortical extract before the stress, the carbohydrate response was within normal limits. The injection of pantothenic acid immediately before the test also improved the response but did not restore it to normal. 

Oat (Avena sativa) extract Pantothenic acid Pyridoxine Riboflavin 5 -phosphate vitamin A determination 1,3-Dichloro-2-propanol vitamin A mfg. 

Phytochemistry Wild carrot seeds contained up to 2.5 % essential oils, which consisted of 17 substances including 1-a-pinene, myrcene, bergamotene, (3-bisabolene, caratol and asarone. Besides essential oil, the seeds contained flavonoids, coumarins, steroidal compounds and fatty oil. The roots also contain essential oils, which consisted of asarone, caratol, bisabolene and 5-9 % carotene. Large amounts of pyrrolidine and daucene were found in essential oils extracted from the herb. In the herb and flowers flavonoids, coumarins, anthocyanins, as well as large amounts of carotenoids, vitamins in the groups B and C, pantothenic acid, anthocyanidin, essential oils, umbelUferone and sugars were found (Ganunerman et al. 1990). 

Coverage includes B vitamins and folate in the context of a historical background, disease, cardiovascular effects and the importance of vitamins in biochemistry as illustrated by a single vitamin. Thereafter there are chapters on the chemistry and biochemistry of thiamine, riboflavin, niacin, pantothenic acid, pyridoxine, biotin, folate and cobalamin. Methodical aspects include characterization and assays of B vitamins and folate in foods of all kinds, dietary supplements, biological fluids and tissues. The techniques cover solid-phase extraction, spectrofluorimetry, mass spectrometry, HPLC, enzymatic assay, biosensor and chemiluminescence. In terms of fimction and effects or... 

Purification of pantothenic acid from an extract is required to measure the total pantothenic acid in foodstuffs (Pakin et al. 2004), while urine samples can... 

Gonthier, A., Fayol, V., Viollet, J., and Hartmann, D. J., 1998a. Determination of pantothenic acid in foods influence of the extraction method. Food Chemistry. 63 287-294. 

Because most B family vitamins are more stable in acidic conditions than in basic conditions, extraction of these vitamins in acidic conditions is preferred. Since pantothenic acid itself is very stable at room temperature, extraction can be kept as simple as possible. Sonication with water or buffer at pH 2-3 is sufficient, even with the extraction of other B family vitamins in mind (Chen and Wolf 2007 Chen et al. 2009). After sonication, centrifugation and/or filtration for sample clean-up is usually sufficient. More complicated sample clean-up procedures, i.e. solid-phase extraction (SPE), are not usually necessary. 

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