Zuo Lihua, Sun Zhi, Hu Yurong, Sun Ya, Xue Wenhua, Zhou Lin, Zhang Jun, Bao Xiaoyue, Zhu Zhenfeng, Suo Guanglu, Zhang Xiaojian
Department of Pharmacy, The First Affiliated Hospital of Zhengzhou University, No.1 Jianshe East Road, Zhengzhou, Henan Province, 450052, PR China.
Department of Pharmacy, The First Affiliated Hospital of Zhengzhou University, No.1 Jianshe East Road, Zhengzhou, Henan Province, 450052, PR China.
J Pharm Biomed Anal. 2017 Apr 15;137:220-228. doi: 10.1016/j.jpba.2017.01.024. Epub 2017 Jan 12.
Xuebijing injection (XBJ) is a traditional Chinese herbal prescription widely used in the treatment of sepsis. Extensive chemical studies revealed that XBJ injection contains amino acids, phenolic acids, flavonoid glycosides, terpeneglycosides and phthalides. In this study, the applicability of ultra high performance liquid chromatography coupled with high resolution hybrid quadruple-orbitrap mass spectrometry (UHPLC-Q-Orbitrap MS) for the simultaneous quantitative analysis of 30 bioactive constituents in XueBiJing injection (XBJ) was investigated. The mass spectrometer was operated in full MS scan mode. The use of 70,000FWHM mass resolution and narrow mass windows (5ppm) could effectively improve the selectivity of the method. Separation was achieved on a Waters ACQUITY UPLC HSS C column (2.1mm×100mm, 1.8μm) with a gradient mobile phase consisting of acetonitrile-water (containing 10mM ammonium acetate) at a flow rate of 0.2mL/min. Satisfactory linearity was achieved within wide linear range and all correlation coefficients (r) of analytes were more than 0.9996. The limits of detection (LODs) were in the range of 0.1180-27.82ng/mL for different analytes. The relative standard deviations (RSDs) of inter- and intra-day precisions were less than 3.0% and the recoveries of the assay were in the range of 98.5%-101.5%. The validated method was successfully applied for simultaneous determination of 30 bioactive compounds in XueBiJing injection from 10 batches samples by UHPLC-Q-Orbitrap MS within 10min. Moreover, the results were evaluated principal component analysis and two compounds might be the most important chemical markers for chemical quality control of XBJ injection. The novel Q-Orbitrap mass spectrometry has been proved to be a very promising and powerful tool for routine screening of bioactive compounds in traditional Chinese medicine injection, ensuring drug safety and public health.
血必净注射液（XBJ）是一种广泛用于治疗脓毒症的传统中药方剂。广泛的化学研究表明，血必净注射液含有氨基酸、酚酸、黄酮苷、萜类糖苷和苯酞类化合物。在本研究中，考察了超高效液相色谱-高分辨率混合四极杆-轨道阱质谱联用技术（UHPLC-Q-Orbitrap MS）同时定量分析血必净注射液（XBJ）中30种生物活性成分的适用性。质谱仪以全扫描模式运行。采用70,000的半高宽质量分辨率和窄质量窗口（5ppm）可有效提高该方法的选择性。在Waters ACQUITY UPLC HSS C柱（2.1mm×100mm，1.8μm）上进行分离，流动相为乙腈-水（含10mM醋酸铵）梯度洗脱，流速为0.2mL/min。在较宽的线性范围内获得了满意的线性关系，各分析物的相关系数（r）均大于0.9996。不同分析物的检测限（LOD）在0.1180 - 27.82ng/mL范围内。日内和日间精密度相对标准偏差（RSD）均小于3.0%，回收率在98.5% - 101.5%范围内。所建立的方法成功应用于UHPLC-Q-Orbitrap MS在10分钟内同时测定10批血必净注射液中的30种生物活性化合物。此外，通过主成分分析对结果进行了评估，两种化合物可能是血必净注射液化学质量控制的最重要化学标志物。新型的Q-Orbitrap质谱已被证明是用于中药注射剂生物活性化合物常规筛选的非常有前景且强大的工具，可确保药物安全和公众健康。
