Analysis of Triterpene Chemical Constituents in Poria cocos 95% Ethanol Extract via UPLC-IT-TOF/MS
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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。 -
关键词:
- 茯苓 /
- 三萜 /
- UPLC-IT-TOF/MS
Abstract:Objective To evaluate the triterpene chemical constituents in Poria cocos 95% ethanol extract by ultra-performance liquid chromatography ion trap time-of-flight mass spectrometry (UPLC-IT-TOF/MS) method. Methods UPLC was performed with the UHPLC XB-C18 and methanoic acid/water (0.05/100, v/v)-acetonitrile solution (gradient elution) was employed with a flow rate of 0.2 mL·min-1. The detection wavelength range was 190~400 nm. Triterpene chemical compositions in Poria cocos were qualitatively analyzed using ESI ion with electrospray positive and negative ions simultaneous detection, sodium trifluoroacetace correction and Shimadzu Composition Formula Predictor software. Results Through the analysis of the multistage tandem mass spectrometry, combined with the application of nitrogen rule and the Scifinder database search, 22 triterpenes with lanostan-8-en, lanostan-7, 9(11)-diene type and other lanostane triterpene were identified. Conclusion This study quickly analyzed the triterpenes in the Poria cocos ethanol extract via LC-MS, which provided the basis for the pharmacodyamic material basis and quality control. -
Key words:
- Poria cocos /
- Triterpene /
- UPLC-IT-TOF/MS
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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-Peroxydehydrotumulosic 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) - 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-dehydropachymic 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]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] -
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