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中國精品科技期刊2020

榛蘑與菌絲體中蜜環菌素的超聲波輔助提取條件優化及其提取物中化合物分析

徐偉 王植朔 王瑞琦 吳凡 梁珊珊 謝紅瑤 張雪

徐偉,王植朔,王瑞琦,等. 榛蘑與菌絲體中蜜環菌素的超聲波輔助提取條件優化及其提取物中化合物分析[J]. 食品工業科技,2022,43(19):298?306. doi:  10.13386/j.issn1002-0306.2022050143
引用本文: 徐偉,王植朔,王瑞琦,等. 榛蘑與菌絲體中蜜環菌素的超聲波輔助提取條件優化及其提取物中化合物分析[J]. 食品工業科技,2022,43(19):298?306. doi:  10.13386/j.issn1002-0306.2022050143
XU Wei, WANG Zhishuo, WANG Ruiqi, et al. Optimization of Ultrasonic-assisted Extraction Conditions of Melleolides from Wild Armillaria mellea and Liquid Culture Mycelium, and Analysis of Their Compounds in Extracts[J]. Science and Technology of Food Industry, 2022, 43(19): 298?306. (in Chinese with English abstract). doi:  10.13386/j.issn1002-0306.2022050143
Citation: XU Wei, WANG Zhishuo, WANG Ruiqi, et al. Optimization of Ultrasonic-assisted Extraction Conditions of Melleolides from Wild Armillaria mellea and Liquid Culture Mycelium, and Analysis of Their Compounds in Extracts[J]. Science and Technology of Food Industry, 2022, 43(19): 298?306. (in Chinese with English abstract). doi:  10.13386/j.issn1002-0306.2022050143

榛蘑與菌絲體中蜜環菌素的超聲波輔助提取條件優化及其提取物中化合物分析

doi: 10.13386/j.issn1002-0306.2022050143
詳細信息
    作者簡介:

    徐偉(1963?),女,博士,教授,研究方向:食品生物技術,E-mail:1436456268@qq.com

  • 中圖分類號: TS201.3

Optimization of Ultrasonic-assisted Extraction Conditions of Melleolides from Wild Armillaria mellea and Liquid Culture Mycelium, and Analysis of Their Compounds in Extracts

  • 摘要: 目的:為分析榛蘑與菌絲體中化合物差異,本研究以東北野生榛蘑子實體和液態菌絲體為研究對象,探究蜜環菌素的最佳提取工藝條件,并對提取物中化合物進行分析。方法:以提取物得率和蜜環菌素含量為指標,采用超聲波細胞破碎輔助石油醚進行萃取,通過單因素實驗和正交試驗對提取工藝參數進行優化;采用超高效液相色譜-串聯質譜(UHPLC-MS/MS)技術,對液態菌絲體和榛蘑子實體中化合物進行分析和鑒定。結果:確定最佳提取工藝條件為料液比1:20 g/mL,超聲波功率300 W,超聲時間20 min,溶劑回流時間50 min,在該條件下液態菌絲體提取物得率為26.8%,蜜環菌素含量為0.74 mg/g;液態菌絲體和榛蘑子實體中化合物分別為305和592個。解析出16種蜜環菌素類和5種疑似蜜環菌素碎片物質,其中菌絲體中含有15種蜜環菌素和3種碎片,總含量分別為2.551和0.588 μg/mL;子實體中含有7種蜜環菌素和4種碎片,總含量分別為2.413和2.124 μg/mL。兩者同有組分有6種蜜環菌素和2種碎片;液態菌絲體獨有9種蜜環菌素和1種碎片為:10-dehydroxymelleoloede、蜜環菌辛素、Arnamiol、蜜環菌壬素、A52a、4-dehydroxyarmillarin、蜜環菌丙素、(4R,5S,7R,9S,13R)-2’,5-epoxy-4-dehydroxyarmillarin、4’-demethoxyarmillaribin以及碎片C;榛蘑子實體獨有1種蜜環菌素和2種碎片為:Melledonal以及碎片D和E。結論:液態菌絲體蜜環菌素類化合物多于子實體中,為開發輔助功能食品及藥物提供參考。
  • 圖  1  樣品制備流程

    Figure  1.  Sample preparation process

    圖  2  榛蘑野生子實體、液態菌絲體及粉末形態

    Figure  2.  Wild fruiting body, liquid mycelium, and powder of Armillaria mellea

    注:A,野生榛蘑子實體;B,野生子實體粉末;C,母體純菌株培養菌絲體;D,液態培養菌絲體粉末。

    圖  3  料液比對菌絲提取物得率及其中蜜環菌素含量的影響

    Figure  3.  Effect of solid-liquid ratio on yield of Armillaria mellea extract and the content of Melleolides

    圖  4  超聲波功率對榛蘑菌絲提取物得率及其中蜜環菌素含量的影響

    Figure  4.  Effect of ultrasonic power on the yield of Armillaria mellea extracts and the content of Melleolides

    圖  5  超聲波時間對榛蘑菌絲提取物得率及其中蜜環菌素含量的影響

    Figure  5.  Effects of ultrasonic time on yield of Armillaria mellea extract and the content of Melleolides

    圖  6  溶劑回流時間對榛蘑菌絲提取物得率及其中蜜環菌素含量的影響

    Figure  6.  Effect of solvent reflux time on yield of Armillaria mellea extract and the content of Melleolides

    圖  7  UHPLC-MS/MS分析榛蘑菌絲體、子實體提取物得到的一級總離子流圖

    Figure  7.  First-order total ion chromatograms obtained from UHPLC-MS/MS analysis of Armillaria mycelium and fruit body extracts

    注:A+,菌絲體正離子模式;A−,菌絲體負離子模式;B+,子實體正離子模式;B−,子實體負離子模式。

    圖  8  蜜環菌素及其碎片結構式

    Figure  8.  Melleolides and its fragment structure

    注:化合物信息詳見表3。

    表  1  L9(34)正交實驗因素水平設計

    Table  1.   L9 (34) Orthogonal experimental design factor level table

    因素因素
    A料液比
    (g/mL)
    B超聲波功率
    (W)
    C超聲波時間
    (min)
    D溶劑回流時間
    (min)
    11:152501530
    21:203002050
    31:253502570
    下載: 導出CSV

    表  2  正交試驗結果

    Table  2.   Orthogonal test results

    實驗號A料液比
    (g/mL)
    B超聲波
    功率(%)
    C超聲波
    時間(min)
    D溶劑回流
    時間(min)
    蜜環菌素
    含量(mg/g)
    111110.235
    212220.615
    313330.250
    421230.480
    522310.340
    623120.315
    731320.280
    832130.300
    933210.265
    K11.10.9950.850.84
    K21.1351.2551.361.21
    K30.8450.830.871.03
    R0.290.4250.490.37
    k10.3666670.3316670.2833330.280000
    k20.3783330.4183330.4533330.403333
    k30.2816670.2766670.2900000.343333
    r0.0966670.1416670.1633330.123333
    因素主次C>B>D>A
    優方案C2B2D2A2
    下載: 導出CSV

    表  3  蜜環菌素類化合物分析結果

    Table  3.   Results of Melleolide compounds analysis

    NO.化合物離子
    模式
    化學式分子量+/-保留時間
    (min)
    菌絲體峰
    面積+/-
    菌絲體相對濃
    度+/-(μg/mL)
    子實體峰
    面積+/-
    子實體相對濃
    度+/-(μg/mL)
    1蜜環菌甲素+C24H30O6414.2042111.9114167090651.3194398727671.584
    2碎片A+C9H10O4182.0580112.9711387606820.439141089780.051
    3蜜環菌癸素+C24H30O6414.2041912.3051062259700.3361593256360.574
    4melleolide B-C24H32O7432.2152912.61154350496.10.17228784600.010
    5蜜環菌丁素+C24H28O5396.1936412.97144566661.50.1413961758.70.014
    610-dehydroxymelleoloede-C24H32O6416.2203514.79442204080.50.13400
    74-Methyoxymelleolide+C24H30O6414.2042111.53827128532.50.086207275710.075
    8碎片B+C9H7ClO3198.0085413.10826164975.60.0834221072.70.015
    9蜜環菌辛素-C24H30O7430.199612.38823292384.80.07400
    10Arnamiol-C24H31ClO6450.181415.102226239580.07200
    11碎片C+C9H10O4182.05812.62220770352.60.06600
    126’-Dechloroarnamial+C24H30O6414.2041812.7719754775.90.063388973850.140
    13蜜環菌壬素-C24H29ClO7464.1609313.50514907736.50.04700
    14A52a-C24H31ClO6450.181613.91292628060.02900
    154-dehydroxyarmillarin+C24H30O5398.2094314.88452258.190.02700
    16蜜環菌丙素+C24H27ClO5430.1549913.1046893854.010.02200
    17(4R,5S,7R,9S,13R)-2’,
    5-epoxy-4-dehydroxyarmillarin
    +C24H28O5396.1936212.6326376315.280.02000
    184’-demethoxyarmillaribin-C23H26O5382.1785414.2132690301.770.00900
    19Melledonal-C23H28O8432.1787511.193004386967.50.016
    20碎片D+C9H9O2149.052632.239005281747561.902
    21碎片E+C9H8O3164.047591.2200432065180.156
    下載: 導出CSV
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    [29] KOBORI H, SEKIYA A, SUZUKI T, et al. Bioactive sesquiterpene aryl esters from the culture broth of Armillaria sp.[J]. Journal of Natural Products,2015,78(1):163?167. doi:  10.1021/np500322t
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  • 收稿日期:  2022-05-17
  • 網絡出版日期:  2022-08-19
  • 刊出日期:  2022-09-23

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