灵硒康口服液中灵芝的鉴别及其灵芝多糖中单糖组成的测定

目的鉴别灵硒康口服液中的灵芝成分及测定灵芝多糖中单糖的组成。方法样品经沉淀、离心、固相萃取(solid-phase extraction, SPE)小柱分离、富集和纯化处理。使用福林酚试剂法、薄层色谱法(thin-layer chromatography, TLC)及高效液相色谱法(high performance liquid chromatography, HPLC)鉴别和测定灵硒康口服液中灵芝及灵芝多糖中单糖的组成。结果福林酚试剂法鉴别了灵芝中水溶性蛋白质。样品及对照药材显蓝色,阴性不显色。薄层色谱法鉴别了灵芝中水溶性糖。样品及对照药材在相同的位置上有2个斑点, 1个橘黄色,1个蓝紫色,阴性不显色。以1-苯基-3-甲基-5-吡唑啉酮(1-phenyl-3-methyl-5-pyrazolone,PMP)试剂柱前衍生化、LichrospherC_(18)柱,紫外检测,等度或梯度洗脱的高效液相色谱法测定灵芝多糖中4种单糖。其中等度洗脱含量(mg/100 mL)分别为:甘露糖1.78,葡萄糖124.45,半乳糖3.86,木糖2.68。梯度洗脱分别为:甘露糖1.63,葡萄糖128.65,半乳糖3.52,木糖2.53。两者测定结果基本相同。结论上述鉴别和含量测定方法专属性强、重现性好、灵敏度高,可管控产品质量。

Identification of Ganoderma Lucidum and determination of monosaccharide composition from Ganoderma Lucidum polysaccharide in Lingxikang oral solution

Objective To identify the Ganoderma Lucidum in Lingxikang oral solution and determine the monosaccharide composition in Ganoderma Lucidum polysaccharide. Methods The samples were treated by precipitation, centrifugation, solid phase extraction column separation, enrichment and purification. The Ganoderma Lucidum ingredient and monosaccharide composition of Ganoderma Lucidum polysaccharide in Lingxikang oral solution were identified and determined by folin-phenol reagent method, thin-layer chromatography (TLC) and high performance liquid chromatography (HPLC). Results Folin-phenol reagent method effectively identified the water soluble protein of Ganoderma Lucidum. The sample and the control medicinal material showed blue color, while negative sample did not show color. TLC method effectively identified water soluble sugar. The sample and the control medicinal material showed 2 spots at the same position, one orange and one blue-purple, while the negative sample did not show color. The content of 4 monosaccharide from Ganoderma Lucidum polysaccharide was determined by HPLC with pre-column derivatization (1-phenyL-3-methyL-5-pyrazoLone, PMP), Lichrospher C18 column, UV detection, isocratic or gradient elution methods. The content (mg/100 mL) of mannose, glucose, galactose and xylose was 1.77, 124.45, 3.67 and 2.66 respectively by isocratic elution, while the content was 1.63, 128.65, 3.62 and 2.53 respectively by gradient elution. The results of the 2 tests were basically the same. Conclusion The above methods have strong specificity, good reproducibility, and high sensitivity, which can be used to ensure the quality of the products.