目的 建立HPLC法對不同培養(yǎng)階段及不同培養(yǎng)基培養(yǎng)的蟬花培養(yǎng)物中麥角甾醇水平進行測定。方法 建立麥角甾醇HPLC檢測方法，應(yīng)用Agilent Eclipse XDB-C₁₈色譜柱（4.6 mm×250 mm，5 μm）；流動相為100%甲醇，等度洗脫；體積流量1.0 mL/min；柱溫35℃；進樣量10 μL；檢測波長281 nm；外標法測定麥角甾醇水平。對液體、固體培養(yǎng)基不同培養(yǎng)階段，假單胞菌選擇培養(yǎng)基（PSA）、沙氏葡萄糖瓊脂培養(yǎng)基（SDAY，1 000 mL中含葡萄糖量分別為20、30、40 g），以及包括江蘇農(nóng)墾、煙農(nóng)24、煙農(nóng)15和濟麥22在內(nèi)的固體培養(yǎng)基培養(yǎng)的蟬花培養(yǎng)物中麥角甾醇的水平進行測定，利用SPSS 16.0統(tǒng)計軟件對測定結(jié)果進行多重統(tǒng)計分析，驗證并找出引起差異的原因。結(jié)果 建立的HPLC法，麥角甾醇分離度良好，精密度、穩(wěn)定性、重復(fù)性的RSD分別為0.30%、0.07%、1.37%，加樣回收率為104.3%，實驗重現(xiàn)性、耐用性良好，均符合要求，可用于蟬花培養(yǎng)物中麥角甾醇的檢測；液-固不同培養(yǎng)階段麥角甾醇生長曲線的表現(xiàn)特征不同；不同培養(yǎng)基培養(yǎng)的蟬花培養(yǎng)物中麥角甾醇水平差異顯著，斜面培養(yǎng)基中，隨著葡萄糖量的增加，麥角甾醇水平增加；固體培養(yǎng)基濟麥22和煙農(nóng)15的麥角甾醇的水平顯著高于煙農(nóng)24和江蘇農(nóng)墾。結(jié)論 液-固培育階段麥角甾醇的生長曲線特征不同，且對于培養(yǎng)基具有一定的選擇性，為人工蟬花培養(yǎng)工藝的確定及優(yōu)化提供了依據(jù)，也為針對性的藥物開發(fā)奠定基礎(chǔ)。

Objective The measurement method was established to determine the content of ergosterol in culture of Cordyceps cicadae, and the content of ergosterol was measured in cicada fruiting bodies for different culture medium and different stages. Methods The HPLC method was applied to determine the content of ergosterol in C. cicadae. Chromatography was carried out on an Agilent Eclipse XDB-C₁₈ column (4.6 mm×250 mm, 5 μm) using isocratic elution program at the volume flow rate of 1.0 mL/min. Mobile phase was 100% methanol, column temperature was 35℃, the sample weight was 10 μL and detection wave length was 281 nm. External standard method was used to calculate the content of ergosterol. The content of ergosterol was measured in C. cicadae fruiting bodies for different culture medium including different slope medium such as Pseudomonas Selection Medium (PSA) and Sabouraud's Glucose Agar Medium (SDAY) containing 20, 30, or 40 g glucose sugar in 1000 mL medium; The different solid medium were Jiangsu state farms, Yannong 24, Yannong 15, and Jimai 22 and different stages in liquid and solid medium. The SPSS 16.0 statistical software was applied to verify and find out the reason causing the difference. Results The separation degree of ergosterol was good, RSD of precision, stability, and repetition was 0.30%, 0.07%, and 1.37% respectly, sample recovery rate (104.3%), reproducibility, and durability of HPLC method have been verified to meet the requirements. It could be used for the detection of ergosterol in C. cicadae. The growth curves of ergosterol in different stages of liquid-solid cultures had different characteristics. The ergosterol level was significantly different in different cultures. The ergosterol levels increased with the increase of amount of glucose in slant culture medium. The ergosterol levels in solid culture medium of Jimai 22 and Yannong 15 were significantly higher than those of Yannong 24 and Jiangsu Agribusiness. Conclusion It has been confirmed that ergosterol has selectivity for the medium. The growth curves of ergosterol in different stages of liquid-solid cultures have different characteristics. This experiment is conducive to optimization of the process of C. cicadae.

藥用蟲生真菌中試及產(chǎn)業(yè)化平臺的建立