Isolation and identification of vitamin D3, 25-hydroxyvitamin D3, 1,25-dihydroxyvitamin D3 and 1,24,25-trihydroxyvitamin D3 in Solanum malacoxylon incubated with ruminal fluid.
It has been shown that Solanum malacoxylon contains 1 alpha,25-dihydroxyvitamin D3-glycoside. The presence of vitamin D3 and 25-hydroxyvitamin D3 has also been suggested. In the present study vitamin D3 and three of its metabolites, including 1 alpha,25-dihydroxyvitamin D3, were detected in plant leaf extracts preincubated with ruminal fluid (SMRF). Extraction of SMRF with non-polar organic solvents and purification of the lipid extract by TLC followed by HPLC yielded nine ultraviolet-absorbing (264 nm) peaks. Four of them comigrated on a Zorbax-Sil HPLC column with synthetic standards of vitamin D3, 25-hydroxyvitamin D3, 1 alpha,25-dihydroxyvitamin D3 and 1,24R,25-trihydroxyvitamin D3, respectively. These compounds were unequivocally identified by means of mass spectrometry. The results confirm that Solanum malacoxylon contains, in addition to 1 alpha,25-dihydroxyvitamin D3, vitamin D3, 25-hydroxyvitamin D3 and possibly other as yet unidentified derivatives. As 1,24,25-trihydroxyvitamin D3 is absent in plant extracts not incubated with ruminal fluid, the data also indicate that rumen microbes may convert 1 alpha,25-dihydroxyvitamin D3 into 1,24,25-trihydroxyvitamin D3.
Skliar
M I
MI
Departamento de Biologia, Universidad Nacional del Sur, Bahia Blanca, Argentina.
Boland
R L
RL
Mourino
A
A
Tojo
G
G
D000818:Animals; D002117:Calcitriol / Q000302:isolation & purification*; D002762:Cholecalciferol / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid; D006887:Hydroxycholecalciferols / Q000302:isolation & purification*; D013058:Mass Spectrometry; D010944:Plants / Q000737:chemistry*; D012417:Rumen; D012756:Sheep
D006887:Hydroxycholecalciferols; D002762:Cholecalciferol; C030049:1,25,26-trihydroxyvitamin D3; D002117:Calcitriol