正交优化冬虫夏草菌种油混悬制剂

Optimization on Oil Suspension Formulation of Cordyceps sinensis Spawn

摘要:

利用油混悬剂独有的剂型特性可提高人工培育冬虫夏草中的接种侵染率, 因而研制高活力冬虫夏草菌种油混悬剂可推动其产业化的发展。根据油混悬剂的特殊性, 对MTT法进行改进及优化, 结果为: 反应时间120 min、反应温度37°C、以甲苯为萃取剂, 活细胞量与琥珀酸脱氢酶活力线性相关。依据该方法筛选制剂中助剂的种类及浓度, 并通过正交试验进一步优化, 获得冬虫夏草菌种油混悬剂高活力配方为: 菌种量0.15 g/mL、硬脂酸铝60 mg/mL、司班-80 50 μL/mL。稳定性试验结果表明, 该制剂在4°C下保

Abstract:

The particularities of oil suspension formulation can raise the invasive rate of Hirsutella sinensis to host of Hepialidae for the commercialization of artificial cultivation of Cordyceps sinensis. So it is important to develop a high cell viability oil suspension formulation of C. sinensis spawn. According to the char-acteristics of the oil suspension formulation, MTT assay is adapted and optimized. The result is as follows: reaction time 120 min, reaction temperature 37°C, methylbenzene as extracting agent, and a positive linear correlation established between active cell weights and cell viability. Varieties and concentrations of assis-tance agents in oil suspension formulation have been selected with the refined MTT assay, and further opti-mized together with cell concentrations through orthogonal experiment. The optimal combination project was obtained, namely, cell concentration 0.15 g/mL, aluminium stearate 60 mg/mL, and SPAN-80 50 μL/mL. Results of stability test on the oil suspension formulation indicate that cell viability can maintain above 90% at 4°C after one month.