DSM Nutritional Products, Research & Development, Analytical Research Center, P.O. Box 2676, CH-4002 Basel, Switzerland.
J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Apr 15;878(13-14):963-8. doi: 10.1016/j.jchromb.2010.02.026. Epub 2010 Feb 26.
A rapid method for quantification of 25-hydroxy vitamin D3 in different swine tissues based on isotope dilution HPLC-MS has been developed and validated. Six times deuterated analyte is used as internal standard. The method is fast and can be performed with only 1g sample. Sample preparation for kidney, liver, muscle and spleen requires only homogenisation and extraction with methanol. An additional enzymatic digest is required for skin, and clean-up of the extract by solid-phase extraction (SPE) is used for adipose tissue and skin. The lower limit of detection varies from 1 ng/g (muscle) to 5 ng/g (adipose and skin). The method has been successfully applied to various tissue samples of pigs fed for 119 days either 2000 IU of vitamin D3 or 50 microg of 25-hydroxy vitamin D3 per kg feed. For animals ingesting 25-OH-D3 supplements the highest tissue contents were observed in the skin (24.8+/-3.5 ng/g), followed by kidney (14.2+/-1.5 ng/g), liver and muscle (5.7+/-0.6 ng/g). The 25-OH-D3 content in the skin was significantly higher in animals ingesting 2000 IU/kg of vitamin D3 (39.5+/-13.4 ng/g). Levels in selected tissues of some animals were below the lower limit of quantification. No measurable amounts of 25-OH-D3 were found in spleen, abdominal fat and subcutaneous fat of the animals of both groups as well as in the liver, kidney and muscle of the animals ingesting 2000 IU/kg of vitamin D3.
已经开发并验证了一种基于同位素稀释 HPLC-MS 的快速定量不同猪组织中 25-羟基维生素 D3 的方法。六氘代分析物用作内标。该方法快速，仅需 1g 样品即可进行。肾脏、肝脏、肌肉和脾脏的样品制备仅需均质化和甲醇提取。皮肤需要额外的酶消化，脂肪组织和皮肤采用固相萃取（SPE）进行提取净化。检测下限从 1ng/g（肌肉）到 5ng/g（脂肪和皮肤）不等。该方法已成功应用于 119 天龄的猪的各种组织样本，这些猪分别用 2000IU 维生素 D3 或 50μg 25-羟基维生素 D3/kg 饲料喂养。对于摄入 25-OH-D3 补充剂的动物，皮肤中的组织含量最高（24.8+/-3.5ng/g），其次是肾脏（14.2+/-1.5ng/g）、肝脏和肌肉（5.7+/-0.6ng/g）。摄入 2000IU/kg 维生素 D3 的动物皮肤中的 25-OH-D3 含量明显更高（39.5+/-13.4ng/g）。两组动物的某些组织中的 25-OH-D3 含量低于定量下限。在两组动物的脾脏、腹部脂肪和皮下脂肪以及摄入 2000IU/kg 维生素 D3 的动物的肝脏、肾脏和肌肉中均未发现可测量量的 25-OH-D3。
