Metabolites of 1 alpha, 25-dihydroxyvitamin D3 in rat bile.

Metabolites of 1 alpha, 25-dihydroxyvitamin D3 in rat bile.
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大鼠胆汁中 1 α, 25-二羟基维生素 D3 的代谢物。

DOI:
10.1021/bi00558a034
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发表时间:
1980
期刊:
影响因子:
2.9
通讯作者:
DeLuca,HF
DeLuca,HF
中科院分区:
生物学3区
文献类型:
--
作者:
Onisko,BL;Esvelt,RP;Schnoes,HK;DeLuca,HF

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材料和方法质谱在AEI Model MS-902质谱仪(Associated Electric Industries,Ltd.,Manchester,England)在70 eV下使用直接探针进行样品引入,并且源温度高于环境温度110-130 ℃。使用沃茨M6000 A型液相色谱(沃茨,Inc.,Melford,MA),配备有450型可变波长检测器。对于SP-HPLC(直相高压液相色谱),使用Li-Chrosorb Si 60(Merck & Co.,股份有限公司、Rahway,NJ)柱[4.6mm(内径)× 250 mm(内径)]。对于RP-HPLC(反相高压液相色谱法),使用Zor-bax-ODS(Du蓬特Instruments,威尔明顿,DE)柱[4.6 mm(内径)× 250 mm(内径)]。除HPLC级己烷(Fischer Chemical Co.,伊塔斯卡,IL)。将用于HPLC的所有溶剂通过0.2 μ过滤器(Millipore Corp.,贝德福德,MA)。除非另有说明,否则通过用吸水器和25-35 ℃浴旋转蒸发除去溶剂。合成的la,25-(OH)2D 3是来自Hoffmann-La Roche(Nutley,NJ)的礼物。合成的柠檬酸甲酯是在本实验室制备的(R。P. Espyruvate,H. F.德卢卡,还有。K. Schnoes,unconfirmed results). 24-1- 3和25-1-0-03的混合物先前已经与另一个项目(Napoli等人,1978年)。通过RP-HPLC将混合物分级,并通过NMR(270 MHz)和质谱确定两种组分的身份。重氮甲烷是由N-甲基-N-亚硝基对甲苯磺酰胺皂化生成的。放射性化学品及放射性测量。如前所述制备la,25-(OH)2 [3a-3 H] D3(28 Ci/mmol)和la,25-(OH)r [26,27- 14 C] D3(48 Ci/mmol)(Espyruvone等人,1979年)。放射性标记的化合物的纯度和真实性通过在SP-HPLC上使用10% 2-丙醇的己烷溶液与合成la,25-(OH)2D 3的共色谱法检查(Jones & DeLuca,1975)。
Materials and MethodsGeneral. Mass spectra were obtained on an AEI Model MS-902 mass spectrometer (Associated Electrical Industries, Ltd., Manchester, England) at 70 eV using a direct probe for sample introduction and a source temperature of 110-130 C above ambient. High-pressure liquid chromatography (HPLC) was performed with a Waters Model M6000A liquid chromatography (Waters, Inc., Melford, MA) equipped with a Model 450 variable-wavelength detector. For SP-HPLC (straight-phase high-pressure liquid chromatography), a Li-Chrosorb Si 60 (Merck & Co., Inc., Rahway, NJ) column [4.6 mm (id) by 250 mm (Z,)] was used. For RP-HPLC (re-versed-phase high-pressure liquid chromatography), a Zor-bax-ODS (Du Pont Instruments, Wilmington, DE) column [4.6 mm (id) by 250 mm (Z,)] was used. All solvents were redistilled before use with the exception of HPLC-grade hexane (Fischer Chemical Co., Itasca, IL). All solvents for HPLC were filtered through 0.2-µ filters (Millipore Corp., Bedford, MA) before use. Unless otherwise noted, solvents were removed by rotary evaporation with a water aspirator and a 25-35 C bath. Synthetic la, 25-(OH) 2D3 was a gift from Hoffmann-La Roche (Nutley, NJ). Synthetic calcitroic acid methyl ester was prepared in this laboratory (R. P. Esvelt, H. F. DeLuca, and. K. Schnoes, unpublishedresults). A mixture of 24-1--3 and 25-1-0-03 had been prepared previously in connection with another project (Napoli et al., 1978). The mixture was fractionated by RP-HPLC, and the identity of the two components was established by NMR (270 MHz) and mass spectrometry. Diazomethane was generated by saponification of ZV-methyl-ZV-nitroso-ptoluenesulfonamide.Radiochemicals and Measurement of Radioactivity. la, 25-(OH) 2 [3a-3H] D3 (28 Ci/mmol) and la, 25-(OH) r [26, 27-14C] D3 (48 Ci/mmol) were prepared as previously described (Esvelt et al., 1979). Purity and authenticity of the radiolabeled compounds were checked by cochromatography with synthetic la, 25-(OH) 2D3 on SP-HPLC using 10% 2-propanol in hexane (Jones & DeLuca, 1975).
大鼠肠道 25-羟基维生素 D3- 和 1α,25-二羟基维生素 D3-24-羟化酶。
DOI: --
发表时间: 1978
影响因子: 4.8
作者:
Rajiv Kumar;H. Schnoes;H. F. Deluca
通讯作者: H. F. Deluca
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DOI: --
发表时间: 1979
期刊: Biochemistry
影响因子: 2.9
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通讯作者: H. DeLuca
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DOI: --
发表时间: 1977
影响因子: 4.8
作者:
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DOI: --
发表时间: 1976
期刊: Biochemistry
影响因子: 2.9
作者:
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通讯作者: H. F. Deluca
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DOI: 10.1042/bj0970408
发表时间: 1965
期刊: The Biochemical journal
影响因子: --
作者:
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通讯作者: E. Holdsworth