目的: 建立太白茶药材高效液相色谱(HPLC)指纹图谱分析方法,并运用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱(UHPLC-Q Exactive Focus MS/MS)法对太白茶中的特征峰进行定性分析。方法: 采用Agilent TC-C18(250 mm×4.6 mm,5 μm)色谱柱,以甲醇(A)-0.1%甲酸水溶液(B)为流动相,梯度洗脱,流速1.0 mL·min-1,柱温30 ℃,检测波长254 nm,进样量10 μL。采用“中药色谱指纹图谱相似度评价系统2012年版”,建立指纹图谱并进行相似度评价。化学成分分析选用UHPLC-Q Exactive Focus MS/MS技术,采用Waters Acquity UPLC BEH C18(50 mm×2.1 mm, 1.7 μm)色谱柱,以乙腈(A)-0.1%甲酸水(B)为流动相,梯度洗脱,流速0.3 mL·min-1,柱温30 ℃;质谱条件为电喷雾离子源(ESI),负离子模式扫描,扫描范围m/z 80~1 200。结果: 建立的太白茶特征指纹图谱中,标定共有峰8个,10批太白茶药材相似度均≥0.9,各批次之间共有峰的相对保留时间RSD均＜0.5%。对8个共有峰中的4个峰进行了结构指认,依次为鳞片衣酸、羊角衣酸、thamnoliadepsides B、坝巴酸。结论: 所建立的HPLC指纹图谱方法稳定可靠,专属性好,结合UHPLC-Q-Exactive Focus-MS/MS的定性分析,可用于太白茶药材的整体质量综合评价。
李莎莎, 李雅娟, 王晓婷, 毛阿娟, 李凡, 李芳, 张红, 王卫锋
. 太白茶HPLC指纹图谱的建立及UHPLC-QExactive Focus MS/MS联用分析*[J]. 药物分析杂志, 2024
, 44(4)
: 585
-593
.
DOI: 10.16155/j.0254-1793.2024.04.05
Objective: To establish a quality assessment method for lichens of Thamnolia subuliformis (Ehrh.) W. Culb. based on HPLC fingerprint, and qualitative analysis of the chemical constituents by ultra-high performance liquid chromatography coupled with quadrupole/electrostatic field orbital trap high resolution mass spectrometry (UHPLC-Q Exactive Focus MS/MS). Methods: Agilent TC-C18(250 mm×4.6 mm, 5 μm) chromatographic column was used, the mobile phase was methol-0.1% phosphoric acid with gradient elution at the flow rate of 1.0 mL·min-1, the column temperature was 30 ℃, the detection wavelength was 254 nm, and the injection volume was 10 μL. HPLC fingerprints of lichens of Thamnolia subuliformis was established. The software of similarity calculation for traditional Chinese medicines fingerprints (version 2012) was used to establish the fingerprinting of lichens of Thamnolia subuliformis. The chemical constituents were analyzed by UHPLC-Q Exactive Focus MS/MS, The chromatographic separation was performed on a Waters Acquity UPLC BEH C18 (50 mm×2.1 mm, 1.7 μm) column with acetonitrile(A)-0.1% formic acid aqueous solution(B) as mobile phase for gradient elution, the flow rate was 0.3 mL·min-1 and the column temperature was 30 ℃.Mass spectrometry was performed using an electrospray ionization and data was collected in negative ion modes in the range of m/z 80-1 200. Results: Eight common peaks were identified from fingerprints of 10 batches of samples. The RSD values of relative retention time of 8 common peaks of chromotograms of samples were all below 0.5% and the similarities were above 0.9. 4 of the identified peaks were further confirmed by UHPLC-Q Exactive Focus MS/MS as squamatic acid, baeomycesic acid, thamnoliadepsides B, barbatinic acid. Conclusion: The established method of fingerprint is stable, reliable, and specific, which can be used for quality evaluation of lichens of Thamnolia subuliformis.
[1] ONUT BI, BENJAMIN M, SCOFIELD DG, et al. Sharing of photobionts in sympatric populations of Thamnolia and Cetraria lichens: evidence from high-throughput sequencing[J]. Sci Rep, 2018, 8(1):4406
[2] 杨美霞, 王立松, 王欣宇. 中国地茶属地衣的分类及地理分布研究[J]. 植物科学学报, 2015, 33(2):133
YANG MX, WANG LS, WANG XY. Taxonomic and geographic study on the lichen genus Thamnolia from China[J]. Plant Sci J, 2015, 33(2):133
[3] 四川省藏药材标准 2014年版[S]. 2014: 247
Standard of Chinese Medicinal Materierials in Sichuan 2014[S]. 2014: 247
[4] 谢宗万. 全国中草药汇编(下)[M]. 北京: 人民卫生出版社, 1978: 527
XIE ZW. The Compilation of National Chinese Herbal Medicine(Vol Ⅱ)[M]. Beijing: People’s Medical Publishing House, 1978: 527
[5] 苗明三, 孙玉信, 王晓田. 中药大辞典[M]. 太原: 山西科学技术出版社, 2017: 730
MIAO MS, SUN YX, WANG XT. Dictionary of Traditional Chinese Medicine[M]. Taiyuan: Shanxi Science & Technology Publishing House, 2017: 730
[6] 陕西省药材标准 2015年版[S]. 2015: 130
Standard of Chinese Medicinal Materierials in Shaanxi 2015[S]. 2015: 130
[7] 任国媛, 郭启新, 王静, 等. 雪地茶甲醇提取物体外抑菌活性及其稳定性研究[J]. 食品工业科技, 2022, 43(1):147
REN GY, GUO QX, WANG J, et al. Antibacterial activity and stability of methanol extract from Thamnolia subuliformis in vitro[J]. Sci Technol Food Ind, 2022, 43(1):147
[8] DENG YX, ZHAO SY, BIAN YG, et al. Anti-inflammatory activity of polysaccharide isolated from Thamnolia subuliformis[J]. J Donghua Univ(Eng Ed), 2017, 34(5):641
[9] YU HY, SHEN X, LIU D, et al. The protective effects of β-sitosterol and vermicularin from Thamnolia vermicularis (Sw.) Ach. against skin aging in vitro[J]. An Acad Bras Cienc, 2019, 91(4):e20181088
[10] YU X, CAI Y, ZHAO X, et al. Investigation of the chemical structure of anti-amyloidogenic constituents extracted from Thamnolia vermicularis[J]. J Ethnopharmacol, 2022, 289: 115059
[11] LI C, GUO XD, LEI M, et al. Thamnolia vermicularis extract improves learning ability in APP/PS1 transgenic mice by ameliorating both Aβ and Tau pathologies[J]. Acta Pharmacol Sin, 2017, 38(1):9
[12] CHOI RY, HAM JR, YEO J, et al. Anti-obesity property of lichen Thamnolia vermicularis extract in 3T3-L1 cells and diet-induced obese mice[J]. Prev Nutr Food Sci, 2017, 22(4):285
[13] SALGADO F, ALBORNOZ L, CORTÉZ C, et al. Secondary metabolite profiling of species of the genus usnea by UHPLC-ESI-OT-MS-MS[J]. Molecules, 2017, 23(1):54
[14] KUMAR K, SIVA B, SARMA VUM, et al. UPLC-MS/MS quantitative analysis and structural fragmentation study of five Parmotrema lichens from the eastern ghats[J]. J Pharm Biomed Anal, 2018, 156: 45
[15] LE PP, LE LAC, LEGOUIN B, et al. In situ DART-MS as a versatile and rapid dereplication tool in lichenology: chemical fingerprinting of Ophioparma ventosa[J]. Phytochem Anal, 2016, 27(6):354
[16] JIN Y, MA Y, XIE W, et al. UHPLC-Q-TOF-MS/MS-oriented characteristic components dataset and multivariate statistical techniques for the holistic quality control of Usnea[J]. RSC Adv, 2018, 8(28):15487
[17] 马英华, 田婷婷, 解伟伟, 等. 采用UHPLC-Triple-TOF-MS法鉴定松萝主要酚酸类化学成分[J]. 中草药, 2016, 47(3):392
MA YH, TIAN TT, XIE WW, et al. Major phenolic acids in Usneae Filum by UHPLC-Triple-TOF-MS[J]. Chin Tradit Herb Drugs, 2016, 47(3):392
