基于UFLC-IT-TOF/MS技术分析不同产地黄芪的化学成分

为研究不同产地黄芪的化学成分差异,制备了甘肃和内蒙古2个产地共15批黄芪药材的水煎液,利用超快速液相色谱-离子阱飞行时间质谱仪(UFLC-IT-TOF/MS)采集15批黄芪药材水煎液化学成分的指纹图谱,进而运用偏最小二乘-判别分析(PLS-DA)和(非)参数检验对数据进行模式识别和统计分析,以变量投影重要性大于1、统计学差异小于0. 05以及倍数变化大于1. 5为阈值,筛选并鉴定其差异成分。结果发现,不同产地的黄芪水煎液中毛蕊异黄酮、芒柄花素、亚油酸等7个化学成分的含量有明显差异,涉及木脂素、黄酮、脂肪酸等4种类别。本研究为黄芪药材的全面质量控制、准确区分产地以及合理种植栽培提供了重要依据。

基于UFLC-IT-TOF MS分析技术的妇科再造丸化学成分研究

大品种妇科再造丸由42味中药组成,对多种妇科疾病具有显著疗效。但其组方复杂,化学成分尚未被充分阐明,质量控制水平较低。该研究建立了一种采用超快速液相色谱-离子阱飞行时间质谱联用技术(UFLC-IT-TOF MS)快速鉴定妇科再造丸化学成分的方法,系统研究了其化学物质基础。采用COSMOSIL C18色谱柱(3.0 mm×150 mm,2.6μm)进行分离,以0.4%乙酸和乙腈为流动相梯度洗脱;采用电喷雾离子源(ESI)在正负离子模式下采集妇科再造丸MS1、MS2和MS3的碎片信息。通过推导质谱裂解规律,与对照品、文献数据和单味药材图谱比对等方式,共鉴定出155种化学成分,包括萜类成分47个,酚及酚酸类成分43个,黄酮类成分39个,生物碱类成分14个,苯酞类成分8个,其他类成分4个,其中133个成分首次在妇科再造丸中发现。该研究进一步阐明了妇科再造丸的化学成分组成,为其质量标准的提高及药效物质的阐明提供了丰富信息。

Identification of major parent compounds and metabolites in bile, plasma and urine of rats after oral administration of Radix Scutellariae extract by UFLC-IT-TOF/MS

Flavonoids are the main bioactive components of Radix Scutellariae, a well-known traditional Chinese medicine. After oral administration of Radix Scutellariae extract （RSE）, the bile, plasma and urine of rats were analyzed to identify the absorbed parent components and metabolites by ultra-fast liquid chromatography coupled with ion trap time-of-flight mass spectrometry （UFLC-IT-TOF/MS）. A total of 36 different flavonoids, including 13 new metabolites, were identified or tentatively characterized in bile, plasma and urine of rats based on their fragmentation patterns. Among them, 16 flavonoids were identified in RSE in vitro, 25 flavonoids in rat bile, 15 flavonoids in rat plasma, and 14 flavonoids in rat urine. The results indicated that glucuronidation, sulfation and methylation were the major metabolic pathways of flavonoids. It is the first time that the metabolism of RSE was studied comprehensively.

Identification and comparative analysis of the major chemical constituents in the extracts of single Fuzi herb and FuziGancao herb-pair by UFLC-IT-TOF/MS

AIM: The aim of this work was to establish a specific and sensitive method to comprehensively investigate and compare chemical constituents of Fuzi-Gancao herb pair(FG), consisting of Aconitum carmichaelii Debeaux(Fuzi, Chinese) and Roast Radix Glycyrrhizae(Glycyrrhiza glabra L., Gancao, in Chinese) and Fuzi alone to explore the underlying interaction mechanism of FG. METHOD: An ultra-fast liquid chromatography-ion trap/time-of-flight mass spectrometry(UFLC/MS-IT-TOF) method using diazepam as internal standard was developed for the identification and semi-quantitative analysis of the phytochemical constituents of Fuzi and FG. Chromatographic separation was achieved on a UFLC column using a gradient program with 40 mmol?L-1 ammonium acetate and acetonitrile as the mobile phase. RESULTS: Fifty-one of the sixty compounds, including forty-five C19-diterpenoid alkaloids and six C20-diterpenoid alkaloids were tentatively identified in the extracts of Fuzi and FG through accurate mass measurements and fragmentation patterns. Comparing the contents of these alkaloids in these two extracts, it was found that the diester-diterpenoid alkaloids(DDAs) and the alkylolamine-diterpenoid alkaloids(ADAs) were increased, while the monoester-diterpenoid alkaloids(MDAs) were decreased in the extracts of FG. CONCLUSION: This work provided comprehensive information for the quality control of Fuzi preparations, and the further investigation on the compatibility mechanisms of FG.

藿香正气滴丸UFLC指纹图谱研究

目的建立藿香正气滴丸（HZDP）的超快速液相色谱（UFLC）指纹图谱,为该制剂的质量控制提供参考依据。方法采用Waters Acquity UPLC BEH C18色谱柱（100 mm×2.1 mm,1.7μm）;流动相为乙腈-0.2%甲酸水溶液,梯度洗脱：0-6 min,10%-15%乙腈;6-9 min,15%-20%乙腈;9-14 min,20%-30%乙腈;14-19 min,30%-50%乙腈;19-21 min,50%-65%乙腈;21-26 min,65%-75%乙腈;26-28 min,75%乙腈;28-30 min,75%-10%乙腈;30-32 min,10%乙腈;体积流量为0.4 m L/min;柱温30℃;检测波长300 nm。采用中药色谱指纹图谱相似度评价系统软件（2004A版）分析11批HZDP样品。结果建立了11批HZDP的对照指纹图谱,确定了18个共有峰;其中13个归属到药材,6、7、9、10、12号共有峰来源于白芷,4、5、8号共有峰来源于陈皮,1、2、11、13号共有峰来源于厚朴,17号共有峰来源于苍术,并通过对照品对照法和超快速液相色谱-离子阱-飞行时间串联质谱仪（UFLC-IT-TOF/MS）鉴定了其化学成分。结论该方法操作简便、准确可靠、重复性好,可为HZDP的物质基础研究和质量控制提供参考。

Influence of ultrafiltration membrane on ophiopogonins and homoisoflavonoids in Ophiopogon japonicus as measured by ultra-fast liquid chromatography coupled with ion trap time-of-flight mass spectrometry

Ultrafiltration is one of the most fascinating technologies, which makes it possible to improve the quality of traditional medicines for application in the pharmaceutical industry. However, researchers have paid little attention to the effect of ultrafiltration membrane on traditional medicines chemical constituents. In this work, Ophiopogon japonicus(L.f) Ker-Gawl. was used as an example to illuminate the influence of ultrafiltration with different material and molecular weight cut-off(MWCO) membrane on natural chemical constituents as measured by ultra-fast liquid chromatography coupled with ion trap time-of-flight mass spectrometry(UFLC-IT-TOF/MS). Our results indicated that ultrafiltration membrane significantly impacted homoisoflavonoids, especially homoisoflavonoids that were almost completely retained on the polyethersulfone(PES) membrane. We also found that the larger number of aglycone hydroxy and sugar moiety in steroid saponins, the higher the transmittance. Furthermore, the passage rate(%) of ophiogenin type saponins was higher than that of others. The possible adsorptive mechanisms were hydrogen bonding, hydrophobic interactions, and benzene ring interaction by π-π stacking. In conclusion, it is crucial to choose appropriate ultrafiltration membrane based on the characteristics of produce products for application of ultrafiltration technique.