Plant Cell Biotechnology Department, Central Food Technological Research Institute, Mysore, India.
J Agric Food Chem. 2010 Sep 22;58(18):9925-30. doi: 10.1021/jf102159j.
The present study reports methylcobalamin in Spirulina platensis using high-performance liquid chromatography (HPLC), thin-layer chromatography (TLC), microbiological assay, chemiluminescence assay, liquid chromatography-mass spectrometry (LC-MS), and tandem mass spectrometry (MS/MS). Extraction of vitamin B12 from S. platensis was carried out without using cyanide. Partial purification was achieved using Amberlite XAD-2 followed by elution with 80% (v/v) methanol. Activated charcoal facilitated removal of impurities in S. platensis extract and in further purification of vitamin B12. The purified fraction was identified to contain methylcobalamin as analyzed by HPLC and TLC. Authenticity of methylcobalamin was further confirmed by LC-MS and MS/MS. Quantitation of methylcobalamin in a test sample of S. platensis biomass was performed using microbiological assay and chemiluminescence assay and was found to be 38.5±2 and 35.7±2 μg/100 g of dry biomass, respectively.
本研究采用高效液相色谱法（HPLC）、薄层层析法（TLC）、微生物测定法、化学发光法、液相色谱-质谱法（LC-MS）和串联质谱法（MS/MS）对螺旋藻中的甲钴胺进行了报告。从螺旋藻中提取维生素 B12 时未使用氰化物。使用 Amberlite XAD-2 进行部分纯化，然后用 80%（v/v）甲醇洗脱。活性炭有助于去除螺旋藻提取物中的杂质，并进一步纯化维生素 B12。通过 HPLC 和 TLC 分析，鉴定纯化部分含有甲钴胺。LC-MS 和 MS/MS 进一步证实了甲钴胺的真实性。使用微生物测定法和化学发光法对螺旋藻生物质的测试样品中的甲钴胺进行定量，结果分别为 38.5±2 和 35.7±2 μg/100 g 干生物质。
