HPLC双波长法测定小柄马勃子实体、液体发酵菌丝中麦角甾醇和麦角甾酮含量
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摘要
建立HPLC双波长法同时测定小柄马勃(Lycoperdon pedicellatus)子实体与液体发酵菌丝中麦角甾醇和麦角甾酮的含量,并比较2种不同药用部位中麦角甾醇和麦角甾酮的含量差异。采用岛津InertsilODS-SP C18柱(4.6 mm×250 mm, 5μm)色谱柱,流动相为甲醇,流速为0.8 mL·min~(-1),检测波长分别为282 nm、 349 nm,柱温为30℃。结果表明,子实体中麦角甾醇和麦角甾酮的进样量分别在0.02μg~0.20μg (r=1)、 6.54 ng~65.40 ng(r=1)范围内线性关系良好。液体发酵菌丝中麦角甾醇和麦角甾酮的进样量分别在0.305μg~3.05μg (r=1)、6.54 ng~65.40 ng (r=1)范围内线性关系良好。因此所建立的双波长测定法准确可靠、重复性良好,可用于小柄马勃及其液体发酵菌丝中麦角甾醇、麦角甾酮化学成分的含量测定,为控制其质量提供参考依据。
The content of ergosterol and ergosterone in the fruiting bodies and the liquid fermentation mycelium of Lycoperdon pedicellatus through an established dual-wavelength HPLC method were determinated, and the content difference of ergosterol and ergosterone in two different medicinal parts were compared. A Shimadzu Inertsil ODS-SP C18 column(4.6 mm×250 mm, 5μm) was used. The mobile phase was methanol and the flow rate was 0.8 mL·min~(-1), the detection wavelength was 282 nm and 349 nm, and the column temperature was 30°C. The injection amount of ergosterol and ergosterone in the fruiting body was linear in the range of 0.02 μg to 0.20 μg(r=1) and 6.54 ng to 65.40 ng(r=1), respectively. The injection amount of ergosterol and ergosterone in the liquid fermentation mycelium was linear in the range of 0.305 μg to 3.05 μg(r=1) and 6.54 ng to 65.40 ng(r=1), respectively. The established dual-wavelength method was accurate and reliable, and it had a good repeatability. It could be used for the determination of ergosterol and ergosterone in the L. pedicellatus and its liquid fermentation mycelium,which provided a reference for the control of its quality.
The content of ergosterol and ergosterone in the fruiting bodies and the liquid fermentation mycelium of Lycoperdon pedicellatus through an established dual-wavelength HPLC method were determinated, and the content difference of ergosterol and ergosterone in two different medicinal parts were compared. A Shimadzu Inertsil ODS-SP C18 column(4.6 mm×250 mm, 5μm) was used. The mobile phase was methanol and the flow rate was 0.8 mL·min~(-1), the detection wavelength was 282 nm and 349 nm, and the column temperature was 30°C. The injection amount of ergosterol and ergosterone in the fruiting body was linear in the range of 0.02 μg to 0.20 μg(r=1) and 6.54 ng to 65.40 ng(r=1), respectively. The injection amount of ergosterol and ergosterone in the liquid fermentation mycelium was linear in the range of 0.305 μg to 3.05 μg(r=1) and 6.54 ng to 65.40 ng(r=1), respectively. The established dual-wavelength method was accurate and reliable, and it had a good repeatability. It could be used for the determination of ergosterol and ergosterone in the L. pedicellatus and its liquid fermentation mycelium,which provided a reference for the control of its quality.
引文
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[10]李思明,冯怡,曾星. HPLC-APCI-MS/M法同时测定猪苓颗粒中麦角甾酮与麦角甾醇的含量[J].药物分析杂志,2014, 34(4):649-653.
[11]赵英永,程显隆,张萍,等. HPLC法测定猪苓中麦角甾酮的含量[J].药物分析杂志, 2009, 29(9):1579-1581.
[12]王丽莹,胡德,汤磊,等. HPLC双波长法同时测定桦褐孔菌子实体中麦角甾醇和麦角甾酮的含量[J].药学研究,2017, 36(9):509-511, 515.
[13] Reeslev M, Miller M, Nielsen KF. Quantifying mold biomass on gypsum board:comparison of ergosterol and beta-Nacetylhexosaminidase as mold biomass parameters[J]. Appl Environ Microbiol, 2003, 69(7):3996-4003.
[14] Martha BA, Mainul H, Raunel T, et al. Effect of pollutants on the ergosterol content as indicator of fungal biomass[J]. J Microbiol Methods, 2002, 50(12):227-236.
[2]邓志鹏,孙隆儒.中药马勃的研究进展[J].中药材, 2006,28(9):996.
[3]宫诗林,徐祉临,张慧洁,等.小柄马勃对慢性肺损伤的治疗及机理研究[J].药学研究, 2017, 3(8):435-438.
[4]左文英,尚孟坤,揣辛桂.脱皮马勃的抗炎、止咳作用观察[J].河南大学学报:医学版, 2004, 23(3):65.
[5]王小沙,杨志旭,李春生,等.复方马勃冲剂的解热降温及抗炎作用研究[J].中国中医急症, 1998, 7(1):32-33.
[6]高云佳,赵庆春,闵鹏,等.脱皮马勃止血有效部位的实验研究[J].解放军药学学报, 2010, 26(6):548-550.
[7]苏明智,罗舟,颜鸣,等.脱皮马勃化学成分的研究[J].中草药, 2012, 43(4):664-666.
[8]崔磊.脱皮马勃化学成分及抗肿瘤活性研究[D].济南:山东大学, 2006.
[9]相聪坤,宿树兰,关胜江,等. HPLC法测定不同产地不同品种马勃药材中麦角甾醇[J].中草药, 2016, 47(8):1397-1400.
[10]李思明,冯怡,曾星. HPLC-APCI-MS/M法同时测定猪苓颗粒中麦角甾酮与麦角甾醇的含量[J].药物分析杂志,2014, 34(4):649-653.
[11]赵英永,程显隆,张萍,等. HPLC法测定猪苓中麦角甾酮的含量[J].药物分析杂志, 2009, 29(9):1579-1581.
[12]王丽莹,胡德,汤磊,等. HPLC双波长法同时测定桦褐孔菌子实体中麦角甾醇和麦角甾酮的含量[J].药学研究,2017, 36(9):509-511, 515.
[13] Reeslev M, Miller M, Nielsen KF. Quantifying mold biomass on gypsum board:comparison of ergosterol and beta-Nacetylhexosaminidase as mold biomass parameters[J]. Appl Environ Microbiol, 2003, 69(7):3996-4003.
[14] Martha BA, Mainul H, Raunel T, et al. Effect of pollutants on the ergosterol content as indicator of fungal biomass[J]. J Microbiol Methods, 2002, 50(12):227-236.