Article(id=1256263562440356063, tenantId=1146029695717560320, journalId=1255847803461844995, issueId=1256263559323967535, articleNumber=null, orderNo=null, doi=10.13346/j.mycosystema.250135, pmid=null, cstr=32115.14.j.mycosystema.250135, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1746633600000, receivedDateStr=2025-05-08, revisedDate=null, revisedDateStr=null, acceptedDate=1749312000000, acceptedDateStr=2025-06-08, onlineDate=1777446173533, onlineDateStr=2026-04-29, pubDate=1771689600000, pubDateStr=2026-02-22, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1777446173533, onlineIssueDateStr=2026-04-29, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1777446173533, creator=13701087609, updateTime=1777446173533, updator=13701087609, issue=Issue{id=1256263559323967535, tenantId=1146029695717560320, journalId=1255847803461844995, year='2026', volume='45', issue='2', pageStart='250058', pageEnd='250280', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1777446172791, creator=13701087609, updateTime=1777447435276, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1256268854674710546, tenantId=1146029695717560320, journalId=1255847803461844995, issueId=1256263559323967535, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1256268854678904851, tenantId=1146029695717560320, journalId=1255847803461844995, issueId=1256263559323967535, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=250135, endPage=, ext={EN=ArticleExt(id=1256263564155826408, articleId=1256263562440356063, tenantId=1146029695717560320, journalId=1255847803461844995, language=EN, title=Chemical composition of Morchella eximia and ergosterol content of its closely related species, columnId=1256263562373226548, journalTitle=Mycosystema, columnName=Research paper, runingTitle=null, highlight=null, articleAbstract=

The chemical constituents of Morchella eximia were isolated by modern chromatographic separation techniques, and the structures were elucidated by nuclear magnetic resonance (NMR), mass spectrometry (MS), etc. Subsequently, a HPLC method was established for determining the ergosterol content in the fruiting bodies of different species of morels. This method was applied to comparatively analyze the ergosterol content in different growth stages (juvenile, mature, and aging stages) of the fruiting bodies of three cultivated species of morels (Morchella importuna, M. sextelata and M. eximia). In total, 13 compounds were isolated and identified from the fruiting bodies of M. eximia, including 5 ergosterol-related compounds. Among these, compounds 2, 9, 10 and 12 were isolated from the genus Morchella for the first time. HPLC analysis revealed significant differences in ergosterol content in the fruiting bodies of different species and growth stages of morels. The highest content was found in the juvenile fruiting bodies of the three species [(1.647±0.013)-(2.289±0.011) mg/g], however the content comparatively decreased in the mature stage. M. sextelata had the highest content [(1.796± 0.008) mg/g], while M. importuna had the lowest [(1.250±0.004) mg/g] in the mature stage. These findings are significant for the further development, utilization, and quality evaluation of morels.

, correspAuthors=Guangbo XIE, authorNote=null, correspAuthorsNote=
*E-mail:
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ORCID: XIE Guangbo (0000-0001-7600-3214)

, authorsList=Yanran YANG, Dinghui LIU, Guangbo XIE, Liyuan XIE), CN=ArticleExt(id=1256263575560139048, articleId=1256263562440356063, tenantId=1146029695717560320, journalId=1255847803461844995, language=CN, title=七妹羊肚菌化学成分及其近缘种麦角甾醇的含量, columnId=1256263563312771301, journalTitle=菌物学报, columnName=研究论文, runingTitle=null, highlight=null, articleAbstract=

首先以七妹羊肚菌Morchella eximia子实体为研究对象,采用硅胶柱色谱、葡聚糖凝胶柱色谱等多种色谱分离方法进行化学成分的分离纯化,并通过核磁共振(NMR)、质谱(MS)等技术鉴定化合物结构。然后以麦角甾醇为测定指标,建立羊肚菌子实体中含量测定的HPLC方法,并采用该方法对3个羊肚菌栽培品种(梯棱羊肚菌、六妹羊肚菌和七妹羊肚菌)及其不同生长期(幼年期、成熟期和老化期)的子实体中麦角甾醇的含量进行比较分析。从七妹羊肚菌子实体中共分离鉴定出13个化合物,包括5个麦角甾类化合物,其中化合物291012为首次从羊肚菌属中分离得到。高效液相色谱分析结果显示不同品种、不同生长期的羊肚菌子实体中麦角甾醇含量差异明显,3个品种的幼年期子实体中的含量最高[(1.647±0.013)-(2.289±0.011) mg/g],成熟期子实体中的含量相应降低,其中六妹羊肚菌最高[(1.796±0.008) mg/g],梯棱羊肚菌最低[(1.250±0.004) mg/g]。本文首次对七妹羊肚菌的化学成分及羊肚菌中麦角甾醇的含量进行了测定,该研究对羊肚菌的进一步开发利用和质量评价都具有重要意义。

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A: Morchella importuna; B: M. sextelata; C: M. eximia. a: Juvenile stage; b: Mature stage; c: Aging stage.

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A:梯棱羊肚菌;B:六妹羊肚菌;C:七妹羊肚菌;a:幼年期;b:成熟期;c:老化期

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A: Ergosterol; B: Test sample.

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A:麦角甾醇;B:供试品

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Theoretical plate number, resolution, and tailoring factor

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编号
Number
保留时间
Retention time/min
峰面积
Peak area
理论塔板数
Theoretical plate number
分离度
Resolution
拖尾因子
Tailoring factor
1 7.416 477 454 6 680 2.719 1.082
2 7.395 476 741 6 701 2.665 1.082
3 7.443 476 236 6 676 2.686 1.082
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理论塔板数、分离度和拖尾因子

, figureFileSmall=null, figureFileBig=null, tableContent=
编号
Number
保留时间
Retention time/min
峰面积
Peak area
理论塔板数
Theoretical plate number
分离度
Resolution
拖尾因子
Tailoring factor
1 7.416 477 454 6 680 2.719 1.082
2 7.395 476 741 6 701 2.665 1.082
3 7.443 476 236 6 676 2.686 1.082
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Accuracy (spiked recovery) test results

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编号
Number
样品称取量
Sample
weight/g
理论含量
Theoretical
content/mg
对照品加入量
Reference standard
weight/mg
实测值
Measured
value/mg
回收率
Recovery
/%
平均回收率
Mean
recovery/%
相对标准偏差
RSD/%
1 0.201 0.332 2 0.43 0.759 6 99.39 99.93 0.91
2 0.202 0.333 9 0.43 0.767 3 100.80
3 0.206 0.340 5 0.43 0.773 7 100.74
4 0.205 0.338 9 0.43 0.764 2 98.93
5 0.207 0.342 2 0.43 0.768 0 99.04
6 0.208 0.343 8 0.43 0.776 9 100.71
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准确度(加样回收率)试验结果

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编号
Number
样品称取量
Sample
weight/g
理论含量
Theoretical
content/mg
对照品加入量
Reference standard
weight/mg
实测值
Measured
value/mg
回收率
Recovery
/%
平均回收率
Mean
recovery/%
相对标准偏差
RSD/%
1 0.201 0.332 2 0.43 0.759 6 99.39 99.93 0.91
2 0.202 0.333 9 0.43 0.767 3 100.80
3 0.206 0.340 5 0.43 0.773 7 100.74
4 0.205 0.338 9 0.43 0.764 2 98.93
5 0.207 0.342 2 0.43 0.768 0 99.04
6 0.208 0.343 8 0.43 0.776 9 100.71
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七妹羊肚菌化学成分及其近缘种麦角甾醇的含量
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杨鄢然 1 , 刘顶慧 1 , 谢光波 1, * , 谢丽源 2
菌物学报 | 研究论文 2026,45(2): 250135
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菌物学报 | 研究论文 2026, 45(2): 250135
七妹羊肚菌化学成分及其近缘种麦角甾醇的含量
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杨鄢然1, 刘顶慧1, 谢光波1, * , 谢丽源2
作者信息
  • 1 电子科技大学生命科学与技术学院,四川 成都 610054
  • 2 四川省食用菌研究所,四川 成都 610066
Chemical composition of Morchella eximia and ergosterol content of its closely related species
Yanran YANG1, Dinghui LIU1, Guangbo XIE1, * , Liyuan XIE2
Affiliations
  • 1 School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu 610054, Sichuan, China
  • 2 Sichuan Institute of Edible Fungi, Chengdu 610066, Sichuan, China
  • ORCID: XIE Guangbo (0000-0001-7600-3214)

出版时间: 2026-02-22 doi: 10.13346/j.mycosystema.250135
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首先以七妹羊肚菌Morchella eximia子实体为研究对象,采用硅胶柱色谱、葡聚糖凝胶柱色谱等多种色谱分离方法进行化学成分的分离纯化,并通过核磁共振(NMR)、质谱(MS)等技术鉴定化合物结构。然后以麦角甾醇为测定指标,建立羊肚菌子实体中含量测定的HPLC方法,并采用该方法对3个羊肚菌栽培品种(梯棱羊肚菌、六妹羊肚菌和七妹羊肚菌)及其不同生长期(幼年期、成熟期和老化期)的子实体中麦角甾醇的含量进行比较分析。从七妹羊肚菌子实体中共分离鉴定出13个化合物,包括5个麦角甾类化合物,其中化合物291012为首次从羊肚菌属中分离得到。高效液相色谱分析结果显示不同品种、不同生长期的羊肚菌子实体中麦角甾醇含量差异明显,3个品种的幼年期子实体中的含量最高[(1.647±0.013)-(2.289±0.011) mg/g],成熟期子实体中的含量相应降低,其中六妹羊肚菌最高[(1.796±0.008) mg/g],梯棱羊肚菌最低[(1.250±0.004) mg/g]。本文首次对七妹羊肚菌的化学成分及羊肚菌中麦角甾醇的含量进行了测定,该研究对羊肚菌的进一步开发利用和质量评价都具有重要意义。

羊肚菌  /  七妹羊肚菌  /  次级代谢产物  /  麦角甾醇  /  含量测定

The chemical constituents of Morchella eximia were isolated by modern chromatographic separation techniques, and the structures were elucidated by nuclear magnetic resonance (NMR), mass spectrometry (MS), etc. Subsequently, a HPLC method was established for determining the ergosterol content in the fruiting bodies of different species of morels. This method was applied to comparatively analyze the ergosterol content in different growth stages (juvenile, mature, and aging stages) of the fruiting bodies of three cultivated species of morels (Morchella importuna, M. sextelata and M. eximia). In total, 13 compounds were isolated and identified from the fruiting bodies of M. eximia, including 5 ergosterol-related compounds. Among these, compounds 2, 9, 10 and 12 were isolated from the genus Morchella for the first time. HPLC analysis revealed significant differences in ergosterol content in the fruiting bodies of different species and growth stages of morels. The highest content was found in the juvenile fruiting bodies of the three species [(1.647±0.013)-(2.289±0.011) mg/g], however the content comparatively decreased in the mature stage. M. sextelata had the highest content [(1.796± 0.008) mg/g], while M. importuna had the lowest [(1.250±0.004) mg/g] in the mature stage. These findings are significant for the further development, utilization, and quality evaluation of morels.

morels  /  Morchella eximia  /  secondary metabolites  /  ergosterol  /  content determination
杨鄢然, 刘顶慧, 谢光波, 谢丽源. 七妹羊肚菌化学成分及其近缘种麦角甾醇的含量. 菌物学报, 2026 , 45 (2) : 250135 - . DOI: 10.13346/j.mycosystema.250135
Yanran YANG, Dinghui LIU, Guangbo XIE, Liyuan XIE. Chemical composition of Morchella eximia and ergosterol content of its closely related species[J]. Mycosystema, 2026 , 45 (2) : 250135 - . DOI: 10.13346/j.mycosystema.250135
羊肚菌Morchella spp. (羊肚菌科Morchellaceae)是一种珍贵的药食两用真菌,其味道鲜美,风味独特,一直被奉为宴席上的美味佳肴(杜习慧等 2014)。羊肚菌营养丰富,研究表明其含有丰富的蛋白质、氨基酸、肽类、脂肪酸、矿物质、维生素和膳食纤维等多种营养成分,特别是钾、锌和硒含量高于多数常见的蘑菇(Gursoy et al. 2009;Tietel & Masaphy 2018)。除了营养价值高以外,羊肚菌还富含多糖、酚类、萜类及甾醇类等多种生物活性化合物,具有机体免疫力调节、抗疲劳、抑制肿瘤、降血脂、抗氧化等多种活性,使其成为一种潜在的功能食品及营养保健品(尚千涵等 2021;Li et al. 2023;Xu et al. 2025)。因此,羊肚菌在国内外市场上备受欢迎,在欧美、东南亚及我国都有较大需求。
羊肚菌野生资源分布较为稀少,采集难度大,难以满足消费者的需求,现有市场供给的羊肚菌主要以人工栽培品种为主(仲启祥等 2024)。我国是羊肚菌人工栽培大国,据统计,2022年我国羊肚菌栽培面积达22 680 hm2,总产量达到了23.95万t (唐禹婷等 2024;魏子涵等 2024)。目前我国羊肚菌规模化、产业化栽培的品种主要包括黑色羊肚菌支系的梯棱羊肚菌Morchella importuna M.Kuo, OʹDonnell & T.J. Volk、六妹羊肚菌M. sextelata M. Kuo和七妹羊肚菌M. eximia Boud. (Liu et al. 2018)。七妹羊肚菌作为重要的羊肚菌栽培品种之一,相关的研究较少,目前主要集中在育种、栽培、菌丝体发酵等方面,关于其化学成分的研究未见报道(马政等 2024;唐禹婷等 2024;Xie L et al. 2024;Zhang et al. 2024;韦春某等 2025)。麦角甾醇是真菌的特征甾醇,存在于绝大多数真菌细胞膜中,它既是真菌生物量的重要标志物之一,也是维生素D2的合成前体(Mattila et al. 2002;Barajas-Aceves et al. 2002;Reeslev et al. 2003)。同时,它自身还具有抗炎、降血脂、免疫调节、抑菌和抗肿瘤等多种生物活性(Rodrigues 2018;程洋洋等 2021;Rangsinth et al. 2023)。
前期我们对人工栽培羊肚菌子实体的化学成分进行了系统的研究(Tu et al. 2021;涂小曼等 2021;Xie G et al. 2024),作为羊肚菌化学成分研究工作的延续,本文对人工栽培的七妹羊肚菌子实体的化学成分进行了研究,同时以麦角甾醇为指标性成分,建立了基于高效液相色谱的含量测定方法,分析比较了人工栽培的梯棱羊肚菌、六妹羊肚菌和七妹羊肚菌在不同生长阶段的麦角甾醇含量。
不同生长阶段的(包括幼年期,成熟期和老化期)梯棱羊肚菌、六妹羊肚菌和七妹羊肚菌子实体均采集自四川省农业科学院现代农业科技创新示范园羊肚菌种植基地(成都市新都区,海拔480 m,104°12′26″E,30°47′13″N),菌种由四川省农业科学院土壤肥料研究所刘理旭助理研究员鉴定。
葡聚糖凝胶(Sephadex LH-20,GE Healthcare),硅胶(100-200目,200-300目,硅胶H,硅胶GF254,青岛海洋化工厂),麦角甾醇标准品(PS011677,成都普思生物科技股份有限公司),甲醇(色谱纯,TEDIA公司),其他化学试剂为分析纯(成都科隆化学品有限公司)。
AV Ⅱ-400核磁共振仪(TMS作为内标, Bruker),Impact Ⅱ ESI-Q-TOF质谱仪(Bruker),Lyovapor L-200冷冻干燥机(Buchi),LC-2050高效液相色谱仪(配DAD检测器、液相处理软件LabSolutions Version 5.110, Shimadzu),BDS Hypersil C18色谱柱(3 μm,100 mm × 3 mm, Thermo),BSA124S电子天平(Sartorius),KQ3200DA超声波清洗机(昆山舒美超声仪器有限公司),XFB-500高速中药粉碎机(吉首中诚制药机械厂)。
将2.8 kg干燥的七妹羊肚菌子实体粉碎后,用95%乙醇室温浸提3次(7 d/次),合并乙醇提取液后减压浓缩至无醇味,得到乙醇浸膏4 L。加入5 L纯净水混合后,用乙酸乙酯萃取3次(9 L/次),合并乙酸乙酯萃取液,减压浓缩回收溶剂后得到黑褐色乙酸乙酯浸膏92 g。
乙酸乙酯浸膏(92 g)经硅胶柱色谱(200-300目,1.6 kg),石油醚-乙酸乙酯(体积比:100:1-1:1)梯度洗脱,得25个组分(Fr.1-Fr.25)。Fr.3 (1.11 g)经硅胶柱色谱(环己烷:乙酸乙酯=70:1)得到5个组分Fr.3.1-Fr.3.5,Fr.3.3 (45 mg)经葡聚糖凝胶柱色谱(Sephadax LH-20,氯仿:甲醇= 2:1,下同)得到化合物1 (40 mg,无色油状物)。Fr.5 (1.37 g)经硅胶柱色谱(环己烷:丙酮=80:1)得3个组分Fr.5.1-Fr.5.3,Fr.5.3 (582 mg)经连续硅胶柱色谱分离及葡聚糖凝胶柱色谱纯化得化合物2 (23 mg,橘黄色固体)。Fr.6 (717.5 mg)经硅胶柱色谱(环己烷:乙酸乙酯=80:1)得到5个组分Fr.6.1-Fr.6.5,Fr.6.3经葡聚糖凝胶柱色谱纯化得到化合物3 (93 mg,无色油状物)。Fr.9有白色固体析出,过滤后以乙酸乙酯为溶剂进行重结晶,得到化合物4 (20 mg,无色针状晶体)。Fr.11也有白色固体析出,过滤后同样采用乙酸乙酯为溶剂进行重结晶,得到化合物5 (20 mg,无色针状晶体)。Fr.14 (2.3 g)经硅胶柱色谱(石油醚:乙酸乙酯=20:1)得5个组分Fr.14.1-Fr.14.5,Fr.14.3 (395 mg)经硅胶柱色谱(二氯甲烷:丙酮=200:1)分离及葡聚糖凝胶柱色谱纯化得化合物6 (5 mg,白色固体);Fr.14.5 (153 mg)经连续性硅胶柱色谱分离及葡聚糖凝胶柱色谱纯化得化合物7 (13 mg,白色固体)。Fr.15 (717.5 mg)经硅胶柱色谱(二氯甲烷:丙酮=200:1)得8个组分Fr.15.1-Fr.15.8,Fr.15.4 (336 mg)经葡聚糖凝胶柱色谱(Sephadax LH-20,氯仿:甲醇=2:1)得到3个组分Fr.15.4.1-Fr.15.4.3,Fr.15.4.1 (191 mg)经硅胶柱色谱(二氯甲烷:丙酮=200:1)分离及葡聚糖凝胶柱色谱纯化得化合物8 (15 mg,无色油状物);Fr.15.7 (76 mg)经硅胶柱色谱(二氯甲烷:丙酮=30:1)及葡聚糖凝胶柱色谱纯化得化合物9 (7 mg,白色固体)。Fr.17 (260.2 mg)经硅胶柱色谱(石油醚:乙酸乙酯=5:1)得3个组分Fr.17.1-Fr.17.3,Fr.17.2 (36 mg)经连续葡聚糖凝胶柱色谱纯化得化合物10 (5 mg,无色胶状物)。Fr.18中有固体析出,过滤后分别收集固体(Fr.18.1)和滤液(Fr.18.2),Fr.18.1 (23 mg)经葡聚糖凝胶柱色谱(Sephadax LH-20,氯仿:甲醇=2:1)纯化分别得化合物11 (15 mg,白色固体)和12 (3 mg,白色固体);Fr.18.2 (717.5mg)经葡聚糖凝胶柱色谱(Sephadax LH-20,氯仿:甲醇=2:1)得4个组分Fr.18.2.1-Fr.18.2.4,Fr.18.2.3 (395 mg)经连续性硅胶柱色谱分离及葡聚糖凝胶柱色谱纯化得化合物13 (5 mg,白色固体)。
色谱柱:Thermo BDS Hypersil C18 (100 mm × 3 mm, 3 μm);柱温:30 ℃;检测波长:282 nm;流动相:甲醇:水(95:5);流速:0.6 mL/min;进样量:5 μL。
精密称取麦角甾醇标准品2.0 mg于10 mL容量瓶中,加入适量甲醇后超声使其充分溶解,加入甲醇定容至10 mL,得浓度为0.2 mg/mL的麦角甾醇对照品溶液,置于4 ℃下避光保存。
梯棱羊肚菌、六妹羊肚菌和七妹羊肚菌不同生长阶段(幼年期、成熟期和老化期)的子实体分别冷冻干燥后,粉碎,过50目筛。分别精密称取不同品种、不同生长阶段的羊肚菌子实体粉末0.4 g,加入16 mL甲醇后称重,然后超声提取45 min,冷却至室温后,加甲醇补足损失的溶剂,过0.45 μm有机滤膜,得供试品溶液,备用。
(1) 理论塔板数、分离度和拖尾因子:精密吸取供试品溶液按1.3.1的色谱条件进行分析,计算理论塔板数、分离度和拖尾因子。
(2) 灵敏度试验:取麦角甾醇对照品母液,稀释配制一系列低浓度的麦角甾醇对照品溶液(250、125、62.5、31.25、15.625、7.812 5 ng/mL),按1.3.1的色谱条件进行分析,测定相应浓度下的信噪比(S/N)。
(3) 重复性试验:取同一浓度的麦角甾醇对照品溶液,按1.3.1的色谱条件下重复进样6次,记录目标峰面积,计算RSD值。
(4) 标准曲线的绘制:取1.3.2项下制备得到的对照品溶液,用甲醇稀释配成系列浓度梯度(5、10、20、40、80、160、200 μg/mL),过0.45 μm有机滤膜后,按1.3.1的色谱条件进行分析,测定峰面积,以麦角甾醇浓度(x,μg/mL)对色谱峰面积(y)绘制标准曲线,得回归方程、相关系数与线性关系。
(5) 精密度试验:按1.3.3所述方法平行制备6份供试品溶液,按1.3.1的色谱条件分别进样分析,记录麦角甾醇峰面积并计算RSD值。
(6) 溶液稳定性试验:按1.3.3所述方法制备供试品溶液,按1.3.1的色谱条件,分别在第0、1、2、4、8、16和24 h时进样分析,记录麦角甾醇峰面积并计算RSD值。
(7) 准确度试验:称取0.2 g左右的羊肚菌子实体粉末6份,向其中加入0.43 mg/mL的麦角甾醇对照品溶液1 mL,按1.3.3所述方法制备供试品溶液,按1.3.1的色谱条件依次进样分析,记录麦角甾醇峰面积并计算回收率及RSD值。
(8) 系统耐用性试验:按照1.3.3所述方法平行制备供试品溶液两份,采用1.3.1的色谱条件,分别改变其柱温(25、30和35 ℃)和流速(0.5、0.6和0.7 mL/min),然后进行供试品溶液的分析,记录理论塔板数、分离度、拖尾因子及峰面积,并计算供试品中麦角甾醇含量及RSD值。对于每次色谱条件的改变(柱温和流速)都绘制相应的标准曲线,用于定量。
分别取梯棱羊肚菌、六妹羊肚菌和七妹羊肚菌不同生长阶段(幼年期、成熟期和老化期)的子实体粉末,按照1.3.3所述方法制备供试品溶液,每组样品设置3个平行试验,按照1.3.1的色谱条件进行分析,记录麦角甾醇峰面积并计算羊肚菌子实体中麦角甾醇的含量。
化合物1,无色油状物,C57H98O6,ESI-MS: 896.765 8 m/z [M+NH4]+1H NMR (400 MHz,CDCl3) δ: 5.37 (12H, m, H-9' × 2, 9'', 10' × 2, 10'', 12' × 2, 12'', 13' × 2, 13''), 5.29 - 5.23 (1H, m, Glycerol H-2), 4.29 (2H, dd, J = 11.9, 4.3 Hz, Glycerol H-1a, H-3a), 4.14 (2H, dd, J = 11.9, 6.0 Hz, Glycero H-1b, H-3b), 2.77 (6H, t, J = 6.5 Hz, H-11' × 2, 11''), 2.31 (2H, t, J = 7.5 Hz, H-2'), 2.31 (4H, t, J = 7.4 Hz, H-2 × 2), 2.08 - 1.97 (12H, m, H-8' × 2, 8'', 14' × 2, 14''), 1.61 (6H, m, H-3' × 2, 3''), 1.21 - 1.40 (42H, m, 3 × 7 × CH2), 0.85 - 0.92 (9H, m, H-18' × 2, 18''); 13C NMR (100 MHz, CDCl3) δ: 173.3, 173.2, 172.8 (C-1' × 2, 1''), 130.2 × 3, 130.0 × 2, 129.9 (C-9' × 2, 9'', 13' × 2, 13''), 128.0 × 3, 127.9 × 3 (C-10' × 2, 10'', 12' × 2, 12''), 68.8 (Glycerol C-2), 62.1 (Glycerol C-1, 3), 34.2, 34.0 × 2 (C-2'', 2' × 2), 31.9, 31.5 × 2 (C-16'', 16' × 2), 29.0 - 29.7 (3 × 6 × CH2), 27.2 × 3 (C-8' × 2, 8''), 25.6 × 3 (C-11' × 2, 11''), 24.8 × 3 (C-3' × 2, 3''), 22.7, 22.5 × 2 (C-17'', 17' × 2), 14.1, 14.0 × 2 (C-18'', 18' × 2)。以上数据与邹建华和戴均贵(2009)的报道一致,故鉴定为三亚油酸甘油酯(glycerol trilinoleate)。
化合物2,橘黄色固体,C54H82O4,ESI-MS: 817.6105 m/z [M+Na]+1H NMR (400 MHz,CDCl3) δ: 5.15 - 5.02 (8H, m, H-6', 10', 14', 18', 22', 26', 30', 34'), 4.93 (1H, m, H-2'), 3.99 (3H, s, 2-OCH3), 3.97 (3H, s, 3-OCH3), 3.18 (2H, d, J = 7.0 Hz, H-1'), 2.11 - 2.02 (16H, m, H-5', 9', 13', 17', 21', 25', 29', 33'), 2.01 (3H, s, 5-CH3), 2.00 - 1.90 (16H, m, H-4', 8', 12', 16', 20', 24', 28', 32'), 1.73 (3H, s, 3'-CH3), 1.67 (3H, s, 35'-CH3), 1.59 × 7, 1.57 (24H, s, 8 × CH3); 13C NMR (100 MHz, CDCl3) δ: 184.7 (C-1), 183.9 (C-4), 144.3 (C-3), 144,2 (C-2), 141.7 (C-6), 138.8 (C-5), 137.6 (C-3'), 135.2, 135.0, 134.9 × 4, 134.8 (C-7', 11', 15', 19', 23', 27', 31'), 124.4, 124.2 × 5, 124.1, 123.8 (C-6', 10', 14', 18', 22', 26', 30', 34'), 118.8 (C-2'), 61.1 × 2 (2-OCH3, 3-OCH3), 39.7 × 8 (C-4', 8', 12', 16', 20', 24', 28', 32'), 26.7 × 6, 26.6, 26.5 (C-5', 9', 13', 17', 21', 25', 29', 33'), 25.7 (35'-CH3), 25.3 (C-1'), 17.6 (C-36'), 16.3 (3'-CH3), 16.0 × 7 (7 × CH3), 11.9 (5-CH3)。以上数据与Kuznetsova et al. (2002)的报道一致,故鉴定为辅酶Q9 (ubiquinone Q9)。
化合物3,无色油状物,C18H32O2,ESI-MS: 279.234 9 m/z [M-H]-1H NMR (400 MHz,CDCl3) δ: 5.44 - 5.27 (4H, m, H-9, 10, 12, 13), 2.77 (2H, t, J = 6.4 Hz, H-11), 2.34 (2H, t, J = 7.4 Hz, H-2), 2.1 - 1.94 (4H, m, H-8, 14), 1.63 (2H, m, H-3), 1.20 - 1.40 (14H, m, H-4, 5, 6, 7, 15, 16, 17), 0.88 (3H, m, H-18); 13C NMR (100 MHz, CDCl3) δ: 180.3 (C-1), 130.2, 130.0 (C-9, 13), 128.0, 127.9 (C-10, 12), 34.1 (C-2), 31.5 (C-3), 29.6, 29.3, 29.2, 29.1, 29.0 (C-4, 5, 6, 7, 15), 27.2, 27.1 (C-8, 14), 25.6 (C-3), 24.6 (C-16), 22.5 (C-17), 14.0 (C-18)。以上数据与Kim et al. (2016)的报道一致,故鉴定为亚油酸(linoleic acid)。
化合物4,无色针状晶体(乙酸乙酯),C28H46O,ESI-MS: 437.182 7 m/z [M+K]+1H NMR (400 MHz,CDCl3) δ: 5.34 (1H, m, H-6), 5.25 - 5.10 (2H, m, H-22, 23), 3.52 (1H, m, H-3), 1.01 (3H, d, J = 6.4 Hz, H-21), 1.00 (3H, s, H-19), 0.91 (3H, d, J = 6.8 Hz, H-28), 0.83 (3H, d, J = 6.4 Hz, H-27), 0.81 (3H, d, J = 6.5 Hz, H-26), 0.69 (3H, s, H-18); 13C NMR (100 MHz, CDCl3) δ: 140.7 (C-5), 135.8 (C-22), 131.7 (C-23), 121.7 (C-6), 71.8 (C-3), 56.8 (C-14), 56.0 (C-17), 50.1 (C-9), 42.8 (C-24), 42.3 (C-13), 42.2 (C-4), 40.1 (C-20), 39.6 (C-12), 37.2 (C-1), 36.5 (C-10), 33.1 (C-25), 31.9 × 2 (C-2, 8), 31.6 (C-7), 28.5 (C-16), 24.2 (C-15), 21.0 (C-11), 20.9 (C-21), 19.9 (C-26), 19.6 (C-27), 19.4 (C-19), 17.6 (C-28), 12.0 (C-18)。以上数据与Şavkıncı et al. (2024)的报道一致,故鉴定为麦角甾-5,22-二烯-3β-醇(ergosta-5,22- dien-3β-ol)。
化合物5,无色针状晶体(乙酸乙酯),C28H44O,ESI-MS: 397.346 5 m/z [M+H]+1H NMR (400 MHz,CDCl3) δ: 5.57 (1H, dd, J = 5.5, 2.1 Hz, H-6), 5.38 (1H, m, H-7), 5.19 (2H, m, H-22, 23), 3.63 (1H, m, H-3), 1.03 (3H, d, J = 6.6 Hz, H-21), 0.94 (3H, s, H-19), 0.91 (3H, d, J = 6.8 Hz, H-28), 0.83 (3H, d, J = 6.4 Hz, H-26), 0.82 (3H, d, J = 6.4 Hz, H-27), 0.63 (3H, s, H-18); 13C NMR (100 MHz, CDCl3) δ: 141.3 (C-8), 139.7 (C-5), 135.5 (C-22), 131.9 (C-23), 119.5 (C-6), 116.2 (C-7), 70.4 (C-3), 55.7 (C-17), 54.5 (C-14), 46.2 (C-9), 42.8 × 2 (C-13, 24), 40.7 (C-4), 40.4 (C-20), 39.0 (C-12), 38.3 (C-1), 37.0 (C-10), 33.0 (C-25), 31.9 (C-2), 28.3 (C-16), 23.0 (C-15), 21.1 × 2 (C-11, 21), 19.9, 19.6 (C-26, 27), 17.6 (C-28), 16.2 (C-19), 12.0 (C-18)。以上数据与Shirane et al. (1996)的报道一致,故鉴定为麦角甾醇(ergosterol)。
化合物6,白色固体,C28H46O,ESI-MS: 416.376 6 m/z [M+NH4]+1H NMR (400 MHz,CDCl3) δ: 5.16 (1H, m, H-7), 4.71, 4.65 (each 1H, br s, H-28), 3.59 (1H, m, H-3), 2.23 (1H, m, H-25), 1.02 (3H, d, J = 6.8 Hz, H-26), 1.02 (3H, d, J = 6.8 Hz, H-27), 0.95 (3H, d, J = 6.4 Hz, H-28), 0.79 (3H, s, H-19), 0.54 (3H, s, H-18); 13C NMR (100 MHz, CDCl3) δ: 156.9 (C-24), 139.5 (C-8), 117.5 (C-7), 105.9 (C-28), 71.1 (C-3), 56.0 (C-17), 55.0 (C-14), 49.4 (C-9), 43.4 (C-13), 40.2 (C-5), 39.5 (C-12), 38.0 (C-4), 37.1 (C-1), 36.2 (C-20), 34.6 (C-22), 34.2 (C-10), 33.8 (C-25), 31.5 (C-2), 31.1 (C-23), 29.6 (C-6), 27.9 (C-16), 22.9 (C-15), 22.0 (C-26), 21.8 (C-27), 21.5 (C-11), 18.8 (C-21), 13.0 (C-19), 11.8 (C-18)。以上数据与 Shirane et al. (1996)的报道一致,故鉴定为麦角甾-7,24(28)-二烯-3β-醇[ergosta-7,24(28)-dien-3β-ol]。
化合物7,白色固体,C28H44O3,ESI-MS: 451.303 9 m/z [M+Na]+1H NMR (400 MHz,CDCl3) δ: 6.50 (1H, d, J = 8.4 Hz, H-7), 6.24 (1H, d, J = 8.5 Hz, H-6), 5.22 (1H, dd, J = 15.2, 7.4 Hz, H-23), 5.14 (1H, dd, J = 15.3, 8.0 Hz, H-22), 3.96 (1H, m, H-3), 0.99 (3H, d, J = 6.6 Hz, H-21), 0.90 (3H, d, J = 6.8 Hz, H-28), 0.88 (3H, s, H-19), 0.83 (3H, d, J = 6.6 Hz, H-27), 0.81 (3H, d, J = 6.6 Hz, H-26), 0.81 (3H, s, H-18); 13C NMR (100 MHz, CDCl3) δ: 135.4 (C-6), 135.2 (C-22), 132.3 (C-23), 130.7 (C-7), 82.1 (C-5), 79.4 (C-8), 66.4 (C-3), 56.2 (C-17), 51.7 (C-14), 51.1 (C-9), 44.5 (C-13), 42.7 (C-24), 39.7 (C-20), 39.3 (C-12), 36.9 (C-4), 36.9 (C-10), 34.7 (C-1), 33.0 (C-25), 30.1 (C-2), 28.6 (C-16), 23.4 (C-11), 20.9 (C-21), 20.6 (C-15), 19.9 (C-27), 19.6 (C-26), 18.2 (C-19), 17.5 (C-28), 12.8 (C-18)。以上数据与Hybelbauerová et al. (2008)的报道一致,故鉴定为过氧麦角甾醇(ergosterol peroxide)。
化合物8,无色油状物,C21H38O4,ESI-MS: 372.308 7 m/z [M+NH4]+1H NMR (400 MHz,CDCl3) δ: 5.41 - 5.28 (4H, m, H-9', 10', 12', 13'), 4.21 - 4.04 (5H, m, Glycerol H-1, 2, 3), 2.77 (2H, t, J = 6.2 Hz, H-11'), 2.34 (2H, t, J = 7.5 Hz, H-2'), 2.04 (4H, m, H-8', 14'), 1.62 (2H, m, H-3'), 1.40 - 1.20 (14H, m, 7 × CH2), 0.89 (3H, t, J = 6.7 Hz, H-18'); 13C NMR (100 MHz, CDCl3) δ: 173.9 (C-1'), 130.2, 130.0 (C-9', 13'), 128.0, 127.9 (C-10', 12'), 68.4 (Glycerol C-2), 65.0 × 2 (Glycerol C-1, 3), 34.1 (C-2'), 31.5 (C-16'), 29.7, 29.6, 29.3, 29.1, 29.1 (C-4', 5', 6', 7', 15'), 27.2 (C-8'), 27.2 (C-14'), 25.6 (C-11'), 24.9 (C-3'), 22.6 (C-17'), 14.1 (C-18')。以上数据与王玉莉等 (2007)的报道一致,故鉴定为2-亚油酸甘油酯(2-linoleoyl glycerol)。
化合物9,白色固体,C28H42O3,ESI-MS: 449.287 4 m/z [M+Na]+1H NMR (400 MHz,CDCl3) δ: 6.59 (1H, d, J = 8.5 Hz, H-7), 6.28 (1H, d, J = 8.5 Hz, H-6), 5.42 (1H, dd, J = 5.9, 1.8 Hz, H-11), 5.27 - 5.10 (2H, m, H-22, 23), 4.00 (1H, m, H-3), 1.09 (3H, s, H-19), 1.00 (3H, d, J = 6.6 Hz, H-21),, 0.91 (3H, d, J = 6.8 Hz, H-28), 0.83 (3H, d, J = 6.6 Hz, H-26), 0.82 (3H, d, J = 6.5 Hz, H-27), 0.73 (3H, s, H-18); 13C NMR (100 MHz, CDCl3) δ: 142.5 (C-9), 135.4 (C-6), 135.1 (C-22), 132.4 (C-23), 130.7 (C-7), 119.7 (C-11), 82.7 (C-5), 78.3 (C-8), 66.3 (C-3), 55.8 (C-17), 48.1 (C-14), 43.6 (C-13), 42.7 (C-24), 41.2 (C-12), 39.9 (C-20), 37.9 (C-10), 36.0 (C-4), 33.0 (C-25), 32.5 (C-1), 30.6 (C-2), 28.6 (C-16), 25.5 (C-19), 20.9 (C-15), 20.7 (C-21), 19.9 (C-26), 19.6 (C-27), 17.5 (C-28), 12.9 (C-18)。以上数据与Hybelbauerová et al. (2008)的报道一致,故鉴定为9,11-dehydroergosterol peroxide((22E,24R)-5α,8α- epidioxyergosta-6,9(11),22-trien-3β-ol)。
化合物10,无色胶状物;1H NMR (400 MHz,CD3OD) δ: 9.70 (1H, s, 1-CHO), 6.90 (2H, s, H-2, 6), 3.88 (3H, s, 4-OCH3); 13C NMR (100 MHz, CD3OD) δ: 192.0 (1-CHO), 151.1 (C-3, 5), 141.2 (C-4), 132.3 (C-1), 108.6 (C-2, 6), 59.3 (4-OCH3)。以上数据与Node et al. (2006)的报道一致,故鉴定为3,5-二羟基-4-甲氧基苯甲醛(3,5-dihydroxy-4-methoxybenzaldehyde)。
化合物11,白色固体,C21H42O4,ESI-MS: 376.340 0 m/z [M+NH4]+1H NMR (400 MHz,CD3OD) δ: 4.16 (1H, dd, J = 11.3, 4.4 Hz, Glycerol H-1a), 4.07 (1H, dd, J = 11.3, 6.2 Hz, Glycerol H-1b), 3.83 (1H, m, Glycerol H-2), 3.56 (2H, m, Glycerol H-3), 2.36 (2H, t, J = 7.4 Hz, H-2'), 1.63 (2H, m, H-3'), 1.30 (28H, br s, 14 × CH2), 0.91 (3H, t, J = 6.8 Hz, H-18'); 13C NMR (100 MHz, CD3OD) δ: 174.1 (C-1'), 69.7 (Glycerol C-2), 65.0 (Glycerol C-1), 62.6 (Glycerol C-3), 33.5 (C-2'), 31.6 (C-16'), 29.4 - 28.8 (12 × CH2), 24.6 (C-3'), 22.3 (C-17'), 13.0 (C-18')。以上数据与Degenhardt & Hofmann (2010)的报道一致,故鉴定为1-硬脂酸甘油酯(1-O-stearoyl glycerol)。
化合物12,白色固体,C8H6O4,ESI-MS: 189.008 9 m/z [M+Na]+1H NMR (400 MHz,CD3OD) δ: 6.41 (1H, br s, H-4), 6.30 (1H, br s, H-6 ), 5.19 (2H, s, H-3); 13C NMR (100 MHz, CD3OD) δ: 171.8 (C-1), 166.0 (C-5), 158.7 (C-7), 151.7 (C-3a), 103.3 (C-7a), 102.3 (C-6), 100.5 (C-4), 69.5 (C-3)。以上数据与刘晓秋等(2003)的报道一致,故鉴定为5,7-二羟基-1(3H)-异苯并呋喃酮(5,7-dihydroxy-1(3H)-isobenzofuranone)。
化合物13,白色固体,C21H38O4,ESI-MS: 372.307 9 m/z [M+NH4]+1H NMR (400 MHz,CDCl3) δ: 5.35 (4H, m, H-9', 10', 12', 13'), 4.20 (1H, dd, J = 11.6, 4.6 Hz, Glycerol H-1a), 4.15 (1H, dd, J = 11.6, 6.1 Hz, Glycerol H-1b), 3.93 (1H, m, Glycerol H-2), 3.69 (1H, dd, J = 11.4, 3.9 Hz, Glycerol H-3a), 3.60 (1H, dd, J = 11.4, 5.7 Hz, Glycerol H-3b), 2.77 (2H, t, J = 6.3 Hz, H-11'), 2.35 (2H, t, J = 7.5 Hz, H-2'), 2.04 (4H, q, J = 6.6 Hz, H-8', 14'), 1.63 (2H, m, H-3'), 1.40 - 1.22 (14H, m, 7 × CH2), 0.89 (3H, t, J = 6.4 Hz, H-18'); 13C NMR (100 MHz, CDCl3) δ: 174.1 (C-1'), 130.2, 130.0 (C-9', 13'), 128.1, 127.9 (C-10', 12'), 70.3 (Glycerol C-2), 65.2 (Glycerol C-1), 63.3 (Glycerol C-3), 34.1 (C-2'), 31.5 (C-16'), 29.7, 29.6, 29.3, 29.1, 29.1 (C-4', 5', 6', 7', 15'), 27.2 (C-8'), 27.2 (C-14'), 25.6 (C-11'), 24.9 (C-3'), 22.6 (C-17'), 14.1 (C-18')。以上数据与Murata et al. (2011)的报道一致,故鉴定为1-亚油酸甘油酯(1-linoleoyl glycerol)。
化合物1-13的结构见图1
不同品种、不同生长阶段的羊肚菌子实体,它们的外部形态特征存在着明显的差异(图2)。(1)幼年期的羊肚菌菌盖长约2 cm,表面呈较浅的羊肚状凹陷,菌柄长约1.5 cm,呈圆柱形,白色或浅黄白色。3个不同品种的羊肚菌菌盖外形及颜色有一定差异,六妹羊肚菌菌盖呈淡黄褐色,顶部呈尖状,七妹羊肚菌菌盖呈浅灰褐色,顶部较圆,梯棱羊肚菌菌盖颜色和七妹羊肚菌相似,但稍深,菌盖顶部钝尖状。(2)成熟期的羊肚菌可上市销售,菌盖长6-7 cm,外观呈明显的羊肚状,凹陷明显,菌柄长约5-6 cm,呈圆柱状,光滑,白色或淡黄白色,下部稍膨大,有褶皱。3个不同品种的羊肚菌菌盖外形及颜色有一定差异,六妹羊肚菌和梯棱羊肚菌菌盖灰褐色,呈锥形,顶部钝尖状,而七妹羊肚菌菌盖整体呈椭圆形,浅灰褐色。(3)老化期的羊肚菌开始产生孢子,菌盖和菌柄均有所膨大,特别是菌柄部分膨大明显,呈不规则圆柱形,菌盖和菌柄的颜色也较成熟期更深。不同品种的羊肚菌幼年期子实体分别采集15朵,而成熟期及老化期子实体则分别采集5朵,冷冻干燥后粉碎,充分混匀,过50目筛备用。
(1) 理论塔板数、分离度和拖尾因子
1.3.1的色谱条件下,供试品溶液中麦角甾醇分析的理论塔板数大于5 000,分离度大于2,拖尾因子小于2,说明麦角甾醇的色谱峰与其他杂质的色谱峰分离程度良好,且色谱峰的对称性良好,均满足2020版中国药典规定的标准(表1图3)。
(2) 灵敏度
灵敏度是用于评价高效液相色谱检测微量物质的能力,通常以信噪比(S/N)来表示。对一系列低浓度麦角甾醇对照品溶液的分析结果显示,在浓度为15.625 ng/mL时,S/N>3,在浓度为62.5 ng/mL时,S/N>10,即麦角甾醇的检测限为15.625 ng/mL,定量限为62.5 ng/mL。
(3) 重复性
重复性是用于评价色谱系统连续进样时响应值的重复性能,针对同一浓度的麦角甾醇对照品溶液,在1.3.1的色谱条件下重复进样6次,分析结果显示6个峰面积数值的RSD为0.076%,远小于2%,说明重复性较好。
(1) 线性关系
以麦角甾醇浓度(x, μg/mL)对色谱峰面积(y)绘制标准曲线,其线性回归方程为y=11 630x- 4 665.4,R2=1,表明麦角甾醇在5-200 μg/mL的浓度范围内,其浓度和峰面积具有良好的线性关系。
(2) 精密度、稳定性和准确度
平行制备的6份供试品溶液在1.3.1的色谱条件下分析,其峰面积RSD为0.367%,小于1%,说明该方法的精密度良好。供试品溶液在24 h内的7次进样分析中,麦角甾醇峰面积RSD为0.086%,远小于2%,说明供试品溶液在24 h内稳定性良好。在加样回收率试验中,回收率在98.93%-100.80%,平均回收率为99.93%,RSD为0.91%,小于2% (表2),说明该方法的准确度良好。
(3) 耐用性
耐用性是指分析方法在参数微小变动时保持结果稳定的能力,本试验对柱温和流速2个参数微小变动所带来的影响进行了考察。柱温设置为25、30和35 ℃,流速设置为0.5、0.6和0.7 mL/min。分析结果显示各次分离的理论塔板数均大于5 000,分离度均大于2,拖尾因子均小于2,表明色谱条件在参数的微小变动下仍然展示了良好的分离效果,系统适应性良好。供试品溶液在不同的流速,不同的柱温下分别测得的麦角甾醇含量值的RSD为0.721%和0.875%,均小于2%,说明该方法在色谱条件出现小的变动时,含量测定结果基本不受影响,显示出较好的耐用性。
采用新建立的分析方法对3个不同品种及其不同生长阶段的羊肚菌子实体中的麦角甾醇进行定量分析,每个样品平行3份,测定结果见图4
七妹羊肚菌是羊肚菌主要栽培品种之一。在对七妹羊肚菌子实体的化学成分研究中,从其乙醇浸膏的乙酸乙酯萃取部位共分离得到13个化合物,包括5个甾醇类化合物,5个长链脂肪酸及其甘油酯类化合物,3个其他类化合物,其中化合物291012为首次从羊肚菌属中分离得到。5个甾醇类化合物均为麦角甾类,此类化合物多具有生物活性。麦角甾醇(ergosterol)是一种广泛存在于真菌细胞膜中的重要甾醇类化合物,其独特的化学结构赋予了细胞膜特定的物理性质,如流动性和通透性,同时还参与调解真菌细胞的信号转导途径,影响细胞的生长、形态和应激反应(Rodrigues 2018)。此外,麦角甾醇还显示出抗炎、降血脂、免疫调节、抑菌和抗肿瘤等多种生物活性(程洋洋等 2021;Rangsinth et al. 2023)。过氧麦角甾醇(ergosterol peroxide)具有独特的5α,8α-过氧桥结构,该结构赋予了它显著的生物活性,特别是广谱的抗肿瘤活性(Merdivan & Lindequist 2017;王璐等 2024)。
梯棱羊肚菌、六妹羊肚菌和七妹羊肚菌均为目前的大田产业化栽培品种。在大田栽培条件下,羊肚菌从子实体的菌盖肉眼可见算起,经过大约25 d的生长周期后,其子实体逐渐进入老化阶段。在这个生长周期中,我们将其分为幼年期、成熟期和老化期3个不同的生长阶段,以便考察羊肚菌子实体在不同生长阶段次生代谢产物的差异。
为了保证羊肚菌子实体内代谢物的稳定,我们采用冷冻干燥的方法对羊肚菌子实体进行干燥,然后粉碎,过50目筛备用。在供试品溶液的制备过程中,我们对提取条件进行了考察,涉及提取方法(超声提取、回流提取、冷浸),提取溶剂(丙酮、甲醇、石油醚、乙醇),提取时间(15、30、45和60 min)和料液比(1:15、1:25、1:40、1:50),最终确定羊肚菌供试品溶液制备的最佳提取条件:以甲醇为提取溶剂,在料液比1:40 (g/mL)下,超声波辅助提取45 min。目前,已有文献报道了不同食用菌中麦角甾醇含量的测定,涵盖了包括段木灵芝、猴头菌、金针菇、暗褐网柄牛肝菌在内的多种食药用菌(戴肖东等 2014;胡代花等 2017;钱正明等 2019;李赛飞等 2024)。在上述供试品溶液制备过程中,尽管提取溶剂、提取时间等参数存在差异,但提取方法无一例外地均采用超声辅助提取法。这表明,该提取方法在麦角甾醇的有效提取方面具有显著优势。
甾醇类化合物,特别是麦角甾醇是食用菌中的主要次生代谢产物,我们对六妹羊肚菌和七妹羊肚菌子实体化学成分的研究也证实了这一点(涂小曼等 2021)。在对羊肚菌不同品种、不同生长期子实体中麦角甾醇含量分析的结果中我们发现,梯棱羊肚菌、六妹羊肚菌和七妹羊肚菌3个品种的幼年期子实体中的麦角甾醇含量最高,分别为(1.647±0.013)、(2.289±0.011)和(1.974±0.002) mg/g。随着子实体的生长,麦角甾醇的含量都呈下降趋势,但品种间麦角甾醇含量的变化规律却不相同。六妹羊肚菌中的麦角甾醇含量呈一直下降的趋势,从幼年期的(2.289±0.011) mg/g降到成熟期的(1.796±0.008) mg/g,进而到老化期的(1.456±0.011) mg/g。相应地,梯棱羊肚菌和七妹羊肚菌中麦角甾醇含量则是先降低然后再小幅增加,梯棱羊肚菌从幼年期的(1.647±0.013) mg/g下降到成熟期的(1.250±0.004) mg/g,然后小幅增加到老化期的(1.533±0.007) mg/g,而七妹羊肚菌则从幼年期的(1.974±0.002) mg/g降到成熟期的(1.690±0.003) mg/g,然后小幅增加到老化期的(1.791±0.009) mg/g。羊肚菌幼年期子实体中的麦角甾醇含量最高,可能与其处于快速生长和细胞膜合成的活跃阶段有关。不过不同食用菌,其子实体中的麦角甾醇含量在不同生长阶段间的变化趋势不尽相同。李赛飞等(2024)对段木灵芝子实体不同生长阶段的麦角甾醇含量进行了分析,结果显示其子实体中麦角甾醇含量呈现连续降低-趋于平稳-小幅上升的动态变化过程,这一变化趋势与梯棱羊肚菌和七妹羊肚菌相似。然而戴肖东等(2014)对猴头菌的研究发现,在其子实体分化至成熟阶段,麦角甾醇含量呈上升趋势,当子实体成熟时含量达到最大值,子实体成熟后麦角甾醇合成降低。猴头菌子实体不同生长阶段麦角甾醇含量的变化趋势和羊肚菌明显不同。成熟期的羊肚菌适于上市销售,对此阶段的3个品种羊肚菌子实体中的麦角甾醇含量比较发现,六妹羊肚菌中的含量最高,为(1.796±0.008) mg/g,然后是七妹羊肚菌的(1.690±0.003) mg/g,梯棱羊肚菌最低,仅为(1.250±0.004) mg/g。以上结果说明,同一食用菌的不同品种间麦角甾醇含量存在差异,这一现象在其他研究中也得到证实。相聪坤等(2016)在对真菌类药材马勃进行麦角甾醇含量测定时发现,其含量在不同品种间存在显著差异,表明麦角甾醇含量受品种影响较大。此外,刘京晶等(2011)在分析不同灵芝品种中麦角甾醇含量时发现,灵芝品种间麦角甾醇含量差异也很明显。在相同段木树种栽培的灵芝品种中,麦角甾醇含量从最高的0.243%到最低的0.093%不等。
本文首先对七妹羊肚菌的化学成分进行了研究,从其成熟子实体中分离得到13个化合物,包括5个甾醇类化合物,5个长链脂肪酸及其甘油酯类化合物,3个其他类化合物,其中化合物291012为首次从羊肚菌属中分离得到。随后,本文采用高效液相色谱法对羊肚菌子实体中麦角甾醇的含量进行了测定,并从品种和生长期2个维度展开了比较分析。测定结果显示,梯棱羊肚菌、六妹羊肚菌和七妹羊肚菌这3个羊肚菌品种,在幼年期子实体中的麦角甾醇含量均处于最高水平,含量范围在(1.647±0.013) mg/g到(2.289±0.011) mg/g之间。随着羊肚菌子实体生长进程的持续推进,麦角甾醇含量呈现出逐渐降低的趋势。其中六妹羊肚菌子实体中麦角甾醇含量下降过程较为平稳,直至老化期;而梯棱羊肚菌和七妹羊肚菌子实体在从成熟期到老化期的过程中,麦角甾醇含量出现了小幅回升的现象。此外,对于作为商品销售的成熟期子实体而言,3个羊肚菌品种间麦角甾醇含量的差异同样明显。本文首次对七妹羊肚菌的化学成分和羊肚菌麦角甾醇的含量测定进行了研究,将为羊肚菌的开发利用及质量评价提供科学依据。
杨鄢然:实验、数据收集及分析、论文撰写;刘顶慧:数据收集及分析;谢光波:实验设计、实验监管及指导、数据分析、论文审阅修订;谢丽源:提供实验材料、实验指导。
该研究不存在任何潜在利益冲突的商业或财务关系。
  • 四川省科技计划项目(2021YFN0094)
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2026年第45卷第2期
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doi: 10.13346/j.mycosystema.250135
  • 接收时间:2025-05-08
  • 首发时间:2026-04-29
  • 出版时间:2026-02-22
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  • 收稿日期:2025-05-08
  • 录用日期:2025-06-08
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Sichuan Science and Technology Program(2021YFN0094)
四川省科技计划项目(2021YFN0094)
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    1 电子科技大学生命科学与技术学院,四川 成都 610054
    2 四川省食用菌研究所,四川 成都 610066

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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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