Rapid analysis of cobalamin coenzymes and related corrinoid analogs by high-performance liquid chromatography

Jacobsen, Donald W.; Green, Ralph; Quadros, Edward V.; Montejano, Yolanda D.

doi:10.1016/0003-2697(82)90363-3

Cited by 56 publications

(14 citation statements)

References 21 publications

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“…Incorporation of label from [ 14 C]methyltetrahydrofolate (methylTHF) and [ 14 C]propionate into cellular macromolecules by cultured fibroblasts, indirect measures of methionine synthase and methylmalonyl-CoA mutase function, respectively, were measured as previously described [5]. Synthesis of cobalamin coenzyme forms in fibroblasts incubated with [ 57 Co]CNCbl bound to transcobalamin in human serum was determined by hot ethanol extraction of cell extracts followed by high performance liquid chromatography [6]. Enzyme-bound and free cobalamin levels in fibroblasts cultured in the presence of transcobalamin-bound [ 57 Co]CNCbl were determined by molecular sieve chromatography on a fast protein liquid chromatography system [7].…”

Section: Studies In Cultured Fibroblastsmentioning

confidence: 99%

Late onset of symptoms in an atypical patient with the cblJ inborn error of vitamin B12 metabolism: Diagnosis and novel mutation revealed by exome sequencing

Kim

Lee

Hwu

et al. 2012

Molecular Genetics and Metabolism

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Section: Studies In Cultured Fibroblastsmentioning

confidence: 99%

Late onset of symptoms in an atypical patient with the cblJ inborn error of vitamin B12 metabolism: Diagnosis and novel mutation revealed by exome sequencing

Kim

Lee

Hwu

et al. 2012

Molecular Genetics and Metabolism

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“…The Cbl forms were separated by HPLC, as described by Jacobsen et al (15 ). The HPLC apparatus (HP 1100; Agilent Technologies) was fitted with a precolumn (SecurityGuard cartridge C18, 4 ϫ 3 mm; Phenomenex) followed by a reversed-phase column (Luna 3 m C18, 150 mm ϫ 4.6 mm; Phenomenex).…”

Section: Hplc Of Released Cblmentioning

confidence: 99%

Assessment of Vitamin B12 Absorption Based on the Accumulation of Orally Administered Cyanocobalamin on Transcobalamin

et al. 2010

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Background: Vitamin B12, or cobalamin (Cbl), is absorbed in the intestine and transported to the cells bound to transcobalamin (TC). We hypothesize that cyanocobalamin (CNCbl) is absorbed unchanged, thereby allowing measurement of the complex of CNCbl bound to TC (TC–CNCbl) to be used for studying the absorption of the vitamin. Methods: TC was immunoprecipitated from serum samples obtained from healthy donors at baseline and at 24 h after oral administration of three 9-μg CNCbl doses over 1 day. Cbl was released by treatment with subtilisin Carlsberg. The different forms of Cbl were isolated by HPLC and subsequently quantified with an ELISA-based Cbl assay. Results: At baseline, the median TC–CNCbl concentration was 1 pmol/L (range, 0–10 pmol/L); the intraindividual variation (SD) was 1.6 pmol/L (n = 31). After CNCbl administration, the TC–CNCbl concentration increased significantly (P = 0.0003, paired t-test), whereas no major changes were observed in any of the other Cbl forms bound to TC (n = 10). Only a moderate additional increase in TC–CNCbl was observed with prolonged (5 days) CNCbl administration (n = 10). We designed an absorption test based on measuring TC–CNCbl at baseline and 24 h after CNCbl intake and established a reference interval for the increase in TC–CNCbl (n = 78). The median absolute increase was 23 pmol/L (range, 6–64 pmol/L), and the relative increase was >3-fold. Conclusions: Our data demonstrate that CNCbl is absorbed unchanged and accumulates on circulating TC. We suggest that measuring TC–CNCbl will improve the assessment of vitamin B12 absorption.

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“…The corrinoids present in the samples or standards were separated with HPLC essentially as described by Jacobsen et al (18 ). The HPLC apparatus (HP 1100, Agilent Technologies) was fitted with a precolumn (SecurityGuard Cartridge, Phenomenex) followed by a reversed-phase column (Luna 3u reversed-phase C18 150 mm ϫ 4.6 mm, Phenomenex), which was maintained at 20°C.…”

Section: Separation Of Kcn-exposed Corrinoidsmentioning

confidence: 99%

A New Principle for Measurement of Cobalamin and Corrinoids, Used for Studies of Cobalamin Analogs on Serum Haptocorrin

Hardlei

Nexø

2009

Clinical Chemistry

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Rapid analysis of cobalamin coenzymes and related corrinoid analogs by high-performance liquid chromatography

Cited by 56 publications

References 21 publications

Late onset of symptoms in an atypical patient with the cblJ inborn error of vitamin B12 metabolism: Diagnosis and novel mutation revealed by exome sequencing

Late onset of symptoms in an atypical patient with the cblJ inborn error of vitamin B12 metabolism: Diagnosis and novel mutation revealed by exome sequencing

Assessment of Vitamin B12 Absorption Based on the Accumulation of Orally Administered Cyanocobalamin on Transcobalamin

A New Principle for Measurement of Cobalamin and Corrinoids, Used for Studies of Cobalamin Analogs on Serum Haptocorrin

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