A method for the simultaneous determination of vitamins D2,D3 and their metabolites in plasma and its application to plasma samples obtained from normal subjects and patients

Masuda, Sonoko; Okano, Toshio; Kobayashi, Tadashi

doi:10.1016/0308-8146(92)90117-k

Cited by 15 publications

(9 citation statements)

References 23 publications

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“…4) Correlation between the assayed values obtained by the proposed method and a previously reported method. Figure 7 shows the correlation between the assayed values obtained by the presently proposed method using calf thymus receptor and the previously reported method using chick intestinal receptor with three steps of HPLC (13). A significant correlation was observed between the two (p<0.01).…”

Section: Methodsmentioning

confidence: 63%

“…A plasma sample containing 2,000 dpm of [3H]-1,25(CH)2D3 was applied to the procedure described in a previous paper (13) and the 1,25(OH)2D fraction purified by application to the 3 steps of HPLC was applied to RRA using either chick intestinal or calf thymus receptor to determine concentrations of 1,25(OH)2D (upper panel). In contrast, the same plasma sample was applied to the procedure described in the text and the 1,25(OH)2D fraction eluted from a C180H column was successively applied to the 3 steps of HPLC described in a previous paper (13). Each 1,25(OH)2D fraction obtained by each step was applied to RRA using either chick intestinal or calf thymus receptor (lower panel).…”

Section: Methodsmentioning

confidence: 99%

“…The results suggest that the amounts of unknown components nonspecifically bound to interfering concomitants besides 1,25(OH)2D in the calf thymus receptor were smaller than those in the chick intestinal receptor. Therefore, a complete clean-up procedure using at least two steps of HPLC (or a Sep-Pak silica cartridge and one step of HPLC) is necessary to eliminate interfering substances in the assay using chick intestinal receptor, A plasma sample containing 2,000dpm of [3H]m1,25(OH)2D3 was applied to the proposed procedure described in the text and the 1,25 (OH)2D fraction eluted from a C18OH column was successively applied to the 3 steps of HPLC described in a previous paper (13). Each profile at each step was described by UV absorption at 265 nm, radioactivity and binding affinity with chick intestinal receptor.…”

Section: Methodsmentioning

confidence: 99%

“…Figure 4 shows comparison of chick embryonal intestinal (9) and calf thymus receptors. A lipid extract from a plasma sample was purified by the three steps of HPLC described in our previous report (13) and applied to both chick intestinal and calf thymus receptors. The assayed values of 1,25 (OH)2D by the two methods were 49.6 and 59.8pg/ml, respectively, which were close to one another.…”

Section: Methodsmentioning

confidence: 99%

“…However, since plasma levels of 1,25(OH)2D are extremely low (normal range: 20-80pg/ml) , it is very difficult to establish a simplified and precise method. Since high-performance liquid chromatography (HPLC) using an ultraviolet (UV) detector and gas-liquid chro matography/mass spectrometry (GC-MS) cannot be applied to the determination of 1,25(OH)2D in plasma because of their low sensitivity (5), radioreceptor assay (RRA) is now most popular for the purpose (6)(7)(8)(9)(10)(11)(12)(13). Cytosol-chromatin receptors (6-13) have been used for RRA.…”

mentioning

confidence: 99%

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Gross and Separative Determination of 1.ALPHA.,25-Dihydroxyvitamin D2 and D3 in Plasma Using Calf Thymus Receptor.

Masuda¹,

Okano²,

Kobayashi³

1993

Journal of Nutritional Science and Vitaminology, J Nutr Sci Vit

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SummaryDetermination of la,25-dihydroxyvitamin D [1,25(OH)2D, gross amounts of 1,25(OH)2D2 and 1,25(OH)2D3] and separative deter mination of 1,25(OH)2D2 and 1,25(OH)2D3 in plasma using calf thymus receptor have been investigated. A lipid extract from 1ml of plasma is applied to a Bond Elut C180H column and an eluate corresponding to 1,25 (OH)2D including both 1,25(OH)2D2 and 1,25(OH)2D3 is applied to calf thymus receptor to assay a gross amount of the two compounds. On the other hand, when separative assay of the two compounds is performed, the 1,25 (OH)2D eluate obtained from the Bond Elut C180H column is further applied to HPLC using a Zorbax SIL column with 5% isopro panol in methylene chloride as a developing solvent to separate the two compounds from one another. The separated eluates are independently applied to the receptor to assay the two compounds. Since less amounts of unknown components non-specifically bound to interfering concomi tants besides 1,25(OH)2D exist in the calf thymus receptor, complicated purification steps to eliminate the concomitants are unnecessary. The detection limit by this method is 1.25 pg/tube which is sensitive enough for a routine method to assay 1,25(OH)2D in plasma. Key Words vitamin D, 1,25-dihydroxyvitamin D, 1,25-dihydroxyvitamin D2, 1,25-dihydroxyvitamin D3, radioreceptor assay, calf thymus receptor Vitamin D (vitamin D is used as a general name for vitamins D2 and D3 and the same nomenclature system is also applied to the metabolites) is metabolized to in the liver and subsequently to la,25-dihy droxyvitamin D (1,25(OH)2D) or 24R,25-dihydroxyvitamin D (24,25(OH)2D) in the kidney (1, 2). 1,25(OH)2D is known as an active form of vitamin D which promotes intestinal calcium absorption and remodeling in bone to keep calcium homostasis. Recent investigations have revealed that 1,25(OH)2D has the activity

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Section: Methodsmentioning

confidence: 63%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Gross and Separative Determination of 1.ALPHA.,25-Dihydroxyvitamin D2 and D3 in Plasma Using Calf Thymus Receptor.

Masuda¹,

Okano²,

Kobayashi³

1993

Journal of Nutritional Science and Vitaminology, J Nutr Sci Vit

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Identification of vitamins A, D and E by particle beam liquid chromatography-mass spectrometry

Careri

Lugari

Mangia

et al. 1995

Fresenius J Anal Chem

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Osteodystrophy in patients with chronic hepatitis and liver cirrhosis

et al. 1996

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Bone mineral density (BMD) of the lumber vertebrae and factors related to bone metabolism were determined in patients with chronic viral hepatitis and patients with liver cirrhosis to clarify correlations between hepatic dysfunction, considered to be one of the causes of hepatic osteodystrophy, and decrease in bone mass. BMD of the second to fourth lumbar vertebrae was determined with a Lunar (Madison, WI, USA) DPX, a dual-energy X-ray absorptiometry diagnostic system. BMD was significantly lowest in patients with liver cirrhosis, followed by patients with chronic hepatitis, and healthy subjects, in this order. There was a significantly positive but weak correlation between albumin and BMD. Levels of 25(OH)D and 1,25(OH)2D were significantly lower in patients with liver cirrhosis than in those with chronic hepatitis. BMD and vitamin D were decreased in all patients whose cholinesterase (ChE) was below 0.3 delta pH. Urinary pyridinoline (Upyr) was significantly higher in the patients with liver cirrhosis, in whom bone mass was decreased, than in the patients with chronic hepatitis, whereas serum osteocalcin levels were distributed in the upper normal range in patients with chronic hepatitis and those with liver cirrhosis. There was a positive correlation between 25(OH)D and serum osteocalcin levels in patients with liver cirrhosis. These results indicate that osteogenesis is decreased and suggest that the decrease in BMD which occurs in viral liver cirrhosis, probably related to decreased, bone formation and slight promotion of bone resorption, reflects deranged hepatic function. This is the first report of Upyr and urinary deoxypyridinoline (UDpyr) determination in patients with liver cirrhosis and patients with chronic hepatitis. The negative correlation of Upyr and UDpyr with ChE is a novel finding.

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A method for the simultaneous determination of vitamins D2,D3 and their metabolites in plasma and its application to plasma samples obtained from normal subjects and patients

Cited by 15 publications

References 23 publications

Gross and Separative Determination of 1.ALPHA.,25-Dihydroxyvitamin D2 and D3 in Plasma Using Calf Thymus Receptor.

Gross and Separative Determination of 1.ALPHA.,25-Dihydroxyvitamin D2 and D3 in Plasma Using Calf Thymus Receptor.

Identification of vitamins A, D and E by particle beam liquid chromatography-mass spectrometry

Osteodystrophy in patients with chronic hepatitis and liver cirrhosis

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