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茯苓95%乙醇提取物三萜类化学成分的UPLC-IT-TOF/MS分析

陈兴龙 张敏 杨琪瑶 罗永谋 曹婷婷 唐丽萍 李勇

陈兴龙, 张敏, 杨琪瑶, 罗永谋, 曹婷婷, 唐丽萍, 李勇. 茯苓95%乙醇提取物三萜类化学成分的UPLC-IT-TOF/MS分析[J]. 昆明医科大学学报, 2021, 42(11): 1-8. doi: 10.12259/j.issn.2095-610X.S20211101
引用本文: 陈兴龙, 张敏, 杨琪瑶, 罗永谋, 曹婷婷, 唐丽萍, 李勇. 茯苓95%乙醇提取物三萜类化学成分的UPLC-IT-TOF/MS分析[J]. 昆明医科大学学报, 2021, 42(11): 1-8. doi: 10.12259/j.issn.2095-610X.S20211101
Xing-long CHEN, Min ZHANG, Qi-yao YANG, Yong-mou LUO, Ting-ting CAO, Li-ping TANG, Yong LI. Analysis of Triterpene Chemical Constituents in Poria cocos 95% Ethanol Extract via UPLC-IT-TOF/MS[J]. Journal of Kunming Medical University, 2021, 42(11): 1-8. doi: 10.12259/j.issn.2095-610X.S20211101
Citation: Xing-long CHEN, Min ZHANG, Qi-yao YANG, Yong-mou LUO, Ting-ting CAO, Li-ping TANG, Yong LI. Analysis of Triterpene Chemical Constituents in Poria cocos 95% Ethanol Extract via UPLC-IT-TOF/MS[J]. Journal of Kunming Medical University, 2021, 42(11): 1-8. doi: 10.12259/j.issn.2095-610X.S20211101

茯苓95%乙醇提取物三萜类化学成分的UPLC-IT-TOF/MS分析

doi: 10.12259/j.issn.2095-610X.S20211101
基金项目: 国家自然科学基金资助项目(31860087);云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目(2019FE001(-271))
详细信息
    作者简介:

    陈兴龙(1989~),男,云南曲靖人,理学博士,讲师,主要从事天然产物化学研究工作

    通讯作者:

    李勇,E-mail:1069237727@qq.com

  • 中图分类号: R284.1

Analysis of Triterpene Chemical Constituents in Poria cocos 95% Ethanol Extract via UPLC-IT-TOF/MS

  • 摘要:   目的  采用超高效液相色谱串联离子阱飞行时间质谱(UPLC-IT-TOF/MS)技术定性分析茯苓95%乙醇提取物的三萜类化学成分。  方法  采用UHPLC XB-C18色谱柱,甲酸/水(0.05/100,v/v)-乙腈作为流动相,梯度洗脱,流速为0.20 mL/min;紫外检测波长范围为190~400 nm。使用ESI离子源,三氟乙酸钠校正,电喷雾正负离子同时检测,Shimadzu Composition Formula Predictor软件定性分析茯苓三萜类化合物。  结果  通过多级质谱数据分析,结合氮规则和Scifinder数据库检索,从茯苓醇提取物中鉴定了22个三萜类成分,结构类型包括羊毛甾-8-烯型三萜、羊毛甾-7,9(11)-二烯型三萜和其他羊毛甾三萜类型。  结论  通过LC-MS快速分析了茯苓醇提取物中三萜类成分,为其药效物质基础和质量控制提供了依据。
  • 图  1  茯苓在LC-MS分析条件下的最大离子流图

    Figure  1.  Base peak chromatogram (BPC) of Poria cocos in both positive (1BPC) and negative ion mode (3BPC)

    图  2  茯苓羊毛甾烷型三萜的化学结构

    Figure  2.  The lanostane triterpenoids chemical structures of Poria cocos

    图  3  茯苓中主要色谱峰的质谱图

    Figure  3.  The mass spectrum of main chromatogram peaks in Poria cocos

    表  1  茯苓中色谱峰的LC/ESI–MSn鉴定结果(1)

    Table  1.   Characterization of the peaks in LC/ESI–MSn chromatogram of Poria cocos (1)

    色谱峰保留时间分子式分子量ESI+ (误差,mDa)ESI (误差,mDa)化合物名称
    1 17.51 C30H48O5 488 [M+Na]+ 511.3328 (−6.6)
    MSn: 511-493.3316 (C30H46O4, +2.8),
    453.3319 (C30H44O3, −4.4)
    [M−H]487.3402 (−2.7) Daedaleanic acid B[10]
    2 19.15 C30H46O6 502 [M+Na]+ 525.3301 (+11.4) [M−H]501.3164 (−5.8) Pinicolic acid F [9]
    3 19.90 C31H48O5 500 MSn: 483.3468 (C31H46O4, −0.1), 465.3319 (C31H44O3, −4.4) [M−H]499.3427 (−0.2) 29α-羟基去氢土莫酸
    29α-Hydroxydehy-dropachymic acid[14]
    4 19.90 C30H48O6 504 [M+Na]+ 527.3331 (−1.2)
    MSn: 527- 495.3077 (C29H44O5, +6.3), 437.2886 (C27H42O3, −14.0)
    [M−H]503.3373 (−0.5) -
    5 21.69 C31H48O6 516 [M+Na]+539.3047 (+6.8)
    MSn: 539- 521.2809 (C31H46O5, −6.5), 507.2633 (C30H44O5, −4.6)
    [M−H]515.3056 (+4.2) 5,​8α-​Dioxy-​3β,​16α-​dihydroxylanosta-​7,
    ​24-​ dien-​21-​oic acid [18]
    6 21.97 C31H46O6 514 [M+H]+515.3331 (−3.6)
    MSn: 515- 497.3360 (C31H44O5, +9.8), 443.2839
    (C31H38O2, −10.6), 433.3267 (C30H40O2, +16.6), 425.2996
    (C28H40O3, −5.4), 415.2933 (C30H38O, −6.2), 385.2397
    (C24H32O4, +2.4), 377.2199 (C25H28O3, +8.8), 335.2455
    (C24H30O, +8.6), 253.1655 (C18H20O, +6.8), 247.1670
    (C16H22O2, −2.3), 229.1583 (C16H20O, −0.4), 201.1704 (C15H20, +6.6)
    [M−H]513.3151 (−7.1)
    MSn: 513- 483.3167 (C31H46O4, −5.6)
    5α,​8α-​过氧化去氢土莫酸
    5α,​8α-​Peroxydehydrotumulos​ic acid[19]
    7 22.54 C31H46O5 498 [M+H]+ 499.3287 (−13.1)
    MSn: 481.3308 ( (C31H44O4, −0.4), 463.3147 (C31H42O3, −6.0)
    [M−H]497.3193 (−7.9)
    MSn: 497- 403.2418 (C24H26O5, −7.2), 389.2435 (C27H34O2, −5.1),
    371.2336(C27H32O, −4.4), 355.2131 (C26H28O, +6.4)
    6α-羟基猪苓酸C
    6α-​Hydroxypolyporenic acid C[11]
    8 23.69 C33H48O8 572 [M+H]+573.3312 (−11.1) [M−H]571.3213 (−6.3) 3-(2-羟基乙酰氧基)- 5α,​8α-​过氧化去氢土莫酸
    3-(2-hydroxyacetoxy)- 5α,​
    8α-​peroxydehydrotumulosic acid [20]
    9 25.78 C31H46O5 498 [M+H]+ 499.3433 (+1.5)
    MSn: 481.3408 ( (C31H44O4, +9.6)- 451.3147 (C30H42O3, −1.4),
    435.2955 (C29H38O3, +6.2), 325.2132 (C22H28O2, −3.0),
    299.2112 (C20H26O2, +10.6)
    [M−H]497.3236 (−3.6) -
    下载: 导出CSV

    表  1  茯苓中色谱峰的LC/ESI–MSn鉴定结果(2)

    Table  1.   Characterization of the peaks in LC/ESI–MSn chromatogram of Poria cocos (2)

    色谱峰保留时间分子式分子量ESI+ (误差,mDa)ESI- (误差,mDa)化合物名称
    10 28.42 C32H50O6 530 [M+Na]+553.3443 (−5.7)
    MSn: 553- 535.3422 (C32H48O5, +2.8), 495.3437
    (C30H48O4, −0.8), 477.3274 (C30H46O3, −6.5), 435.3364
    (C28H44O2,+13.0), 353.2548 (C22H34O2, +9.7)
    [M−H]529.3461 (−7.4) 3-乙酰氧基-16α,26-二羟基-羊毛甾-8,24-二烯-21-酸
    3-​ Acetoxy-​16α,​26-​dihydroxy-lanosta-8,24-dien-21-oic acid[10]
    11 29.01 C33H50O6 542 MSn: 525.3513 (C33H48O5, −6.2)- 507.3459
    (C33H46O4, −1.0), 465.3365 (C31H44O3, +0.2),
    447.3544 (C32H46O, −7.7)
    [M−H]541.3460 (−7.5)
    6α-羟基去氢茯苓酸
    6α-​Hydroxy-dehydropachym​ic acid[13]
    12
    31.15 C31H48O4 484 [M+Na]+485.3592 (−3.3)
    MSn: 485- 467.3492 (C31H46O3, −2.9), 449.3408 (C31H44O2, −0.6),
    311.2388 (C22H30O, +1.9), 293.2259 (C22H28, −0.5)
    [M−H]483.3413 (−6.7)
    MSn: 483- 437.3393 (C30H46O2, −3.2), 421.2978 (C29H42O2, −13.4)
    3β,15α-二羟基-羊毛甾-7,9(11),24-三烯-21-酸
    3β,15α-dihydroxylanosta-7,9(11),24-trien-21-oic acid[15]
    13

    30.27 C31H50O4 486 [M+H]+ 487.3722 (−6.0)
    MSn: 487- 469.3719 (C31H48O3, +4.3),
    451.3553 (C31H46O2, −1.8), 343.2701
    (C23H34O2, +6.9), 313.2464 (C22H32O, −6.2),
    295.2405 (C22H30, −1.5)
    [M−H]485.3570 (−6.6)
    MSn: 483- 423.3371 (C29H44O2,+10.2)
    土莫酸
    Tumulosic acid[10-11]
    14 31.74 C31H46O5 498 [M+H]+ 499.3368 (−5.0)
    MSn: 499- 481.3292 (C31H44O4, −2.0), 463.3291 (C31H42O3, +8.4),
    421.3024 (C29H40O2, −7.7), 325.2198 (C22H28O2, +3.8), 307.2102 (C22H26O, +4.6)
    [M−H]497.3199 (−7.3)
    MSn: 497- 425.2942 (C31H38O, +9.2)
    29-羟基猪苓酸 C
    29-​Hydroxypolyporenic acid C[16]
    15 33.26 C33H52O6 544 [M+H]+ 545.3733 (−10.4)
    MSn: 545- 527.3696 (C33H50O5, −3.5), 451.3097 (C31H46O2, +2.6), 433.3267 (C31H44O, −19.8), 295.2433 (C22H30,+2.3)
    [M−H]543.3623 (−6.8)
    MSn: 543- 467.3227 (C31H48O3, +9.6)
    25-羟基茯苓酸
    25-​Hydroxypachimic acid[11]
    16 33.57 C31H46O4 482 [M+H]+ 483.3420 (−4.9)
    MSn: 483- 465.3327 (C31H44O3, −3.6), 447.3246 (C31H42O2, −1.2), 309.2214 (C22H28O, +0.1)
    [M−H]481.3266 (−5.7)
    MSn: 481- 311.1817 (C21H28O2, −14.0)
    猪苓酸 C
    Polyporenic acid C[11]
    17 34.28 C31H48O4 484 [M+H]+ 485.33637 (+1.2)
    MSn: 483- 467.3486 (C31H46O3, −3.4), 449.3532 (C31H44O2, +11.8), 311.2343 (C22H30O, −2.6), 293.2330 (C22H28,+6.6)
    [M−H]483.3378 (−10.2)
    MSn:
    ​3-表去氢土莫酸
    3-​epi-​Dehydrotumulosic acid acid[11]
    下载: 导出CSV

    表  1  茯苓中色谱峰的LC/ESI–MSn鉴定结果(3)

    Table  1.   Characterization of the peaks in LC/ESI–MSn chromatogram of Poria cocos (3)

    色谱峰保留时间分子式分子量ESI+ (误差,mDa)ESI (误差,mDa)化合物名称
    18 34.89 C31H48O4 484 [M+H]+ 485.3589 (−3.6)
    MSn: 483- 467.3526 (C31H46O3, +0.6), 449.3377 (C31H44O2, −3.7), 311.2408 (C22H30O, +3.9), 293.2265 (C22H28, 0.1)
    [M−H]483.3406 (−7.4)
    MSn: 481- 337.2465 (C24H34O, −7.2)
    去氢土莫酸
    Dehydrotumulosic acid[17]
    19 38.63 C33H50O5 526 [M+H]+ 527.3671(−6.0)
    MSn: 527- 509.3564 (C33H48O4, −6.1), 449.3398 (C31H44O2, −1.6), 293.2240 (C22H28, −2.4)
    [M−H]525.3493 (−9.2)
    MSn:
    3-表去氢茯苓酸
    3-​epi-​Dehydropachymic acid[14]
    20 39.64 C32H50O5 514 [M+H]+ 515.3708(−2.3)
    MSn: 515- 497.3555 (C32H46O4, −7.0), 437.3398 (C30H44O2, −1.6), 295.2422 (C22H30,+0.2)
    [M−H]513.3532 (−5.3)
    3-O-乙酰基-16α-羟基松苓酸
    3-​O-​Acetyl-​16α-​hydroxytrametenolic acid[12]
    21 40.64 C33H50O5 526 [M+H]+ 527.3697 (−3.4)
    MSn: 527- 509.3594 (C33H48O4, −3.1), 449.3446 (C31H44O2, +3.2), 353.2501 (C24H32O2, +2.6),
    293.2254 (C22H28, −1.0)
    [M−H]525.3511 (−7.4)
    MSn: 525- 479.3471 (C32H48O3, −6.0), 465.3351 (C31H46O3, −2.3), 355.2295 (C23H32O3, +1.6),
    去氢茯苓酸dehydropachymic acid[11]
    22 41.87 C33H52O5 528 [M+H]+ 529.3820 (−6.8)
    MSn: 529- 511.3735 (C33H50O4, −4.7), 451.3599 (C31H46O2, −3.0), 295.2419 (C22H30, −0.1)
    [M−H]527.3666 (−7.6)
    MSn: 527- 465.3330 (C31H46O3, −4.4)
    茯苓酸
    Pachymic acid[10-11]
    23 45.90 C30H46O3 454 [M+H]+ 455.3532 (+1.2)
    MSn: 455- 437.3457 (C30H44O2, +4.3), 311.2370 (C22H30O, +0.1), 295.2381 (C22H30, −3.9)
    [M−H]453.3342 (−3.2) 3-羟基-羊毛甾-7,9(11),24-三烯-21酸
    Dehydrotrametenolic acid[12]
    24 47.30 C30H48O3 456 [M+H]+ 457.3663 (−1.3)
    MSn: 457- 439.3502 (C30H46O2, −6.9), 313.2445 (C22H32O, −8.1), 295.2463 (C22H30, +4.3)
    [M−H]455.3471 (−6.0) 3-氢化松苓酸
    trametenolic acid[13]
    下载: 导出CSV
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出版历程
  • 收稿日期:  2021-09-25
  • 网络出版日期:  2021-11-15
  • 刊出日期:  2021-11-30

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