Yang Shuang, DU Qian-Qian, Yue Qian-Xia, Sun Ye-Fen, Jin Chuan-Shan, Zhang Wei, Ma Lei, Liu Cong-Bin, Xie Dong-Mei
Anhui University of Chinese Medicine Hefei 230012, China.
Anhui University of Chinese Medicine Hefei 230012, China Institute of Chinese Medicine Resources Protection and Development, Anhui Academy of Traditional Chinese Medicine Hefei 230012, China.
Zhongguo Zhong Yao Za Zhi. 2023 Feb;48(3):715-724. doi: 10.19540/j.cnki.cjcmm.20220618.201.
In this study, an established ultra-high performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) method was combined with multivariate statistical analysis to investigate the commonality and difference of main chemical components in the medicinal parts of Paeonia lactiflora from different cultivars; in addition, a high performance liquid chromatography(HPLC) method was established to simultaneously determine the content of eight active components in Paeoniae Radix Alba. Non-targeted analysis was carried out by UPLC-Q-TOF-MS on a Waters ACQUITY UPLC BEH C_(18)(2.1 mm×100 mm, 1.7 μm) column with a gradient elution of 0.1% aqueous formic acid(A)-acetonitrile(B) as the mobile phase at a flow rate of 0.2 mL·min(-1). The column temperature was 30 ℃, and an electrospray ionization source was used to acquire mass spectrometry data in positive and negative ion modes. According to the accurate molecular weight and fragment ion information provided by multi-stage mass spectrometry and by comparison with reference substances and literature reports, thirty-six identical components were identified in Paeoniae Radix Alba from different cultivars with positive and negative ion modes. In the negative ion mode, two groups of samples were well separated; specifically, seventeen components with significant differences in content were screened and identified, and one component unique in "Bobaishao" was obtained. Quantitative analysis was conducted by high-performance liquid chromatography(HPLC) on an Agilent HC-C_(18)(4.6 mm×250 mm, 5 μm) column with a gradient elution of 0.1% aqueous phosphoric acid(A)-acetonitrile(B) as the mobile phase at a flow rate of 1.0 mL·min(-1). The column temperature was 30 ℃ and the detection wavelength was at 230 nm. An HPLC method was developed for the simultaneous determination of eight active components(gallic acid, oxypaeoniflorin, catechin, albiflorin, paeoniflorin, galloylpaeoniflorin, 1,2,3,4,6-O-pentagalloylglucose, benzoyl-paeoniflorin) in Paeoniae Radix Albaa from different cultivars. Satisfactory linearity was achieved within the investigated linear ranges and with fine coefficients(r>0.999 0), and the methodological investigation showed that the method had good precision, repeatability and stability. The mean recoveries were 90.61% to 101.7% with RSD of 0.12% to 3.6%(n=6). UPLC-Q-OF-MS provided a rapid and efficient qualitative analytical method for the identification of the chemical components in Paeoniae Radix Alba, and the developed HPLC method was simple, rapid and accurate, which could provide a scientific basis for the evaluation of the germplasm resources and herbal quality of Paeoniae Radix Alba from different cultivars.
本研究将已建立的超高效液相色谱-四极杆-飞行时间质谱联用(UPLC-Q-TOF-MS)方法与多元统计分析相结合,以研究不同栽培品种芍药药用部位主要化学成分的异同;此外,建立了高效液相色谱(HPLC)法同时测定白芍中8种活性成分的含量。采用UPLC-Q-TOF-MS在Waters ACQUITY UPLC BEH C₁₈(2.1 mm×100 mm,1.7 μm)色谱柱上进行非靶向分析,以0.1%甲酸水溶液(A)-乙腈(B)为流动相进行梯度洗脱,流速为0.2 mL·min⁻¹。柱温为30℃,采用电喷雾电离源在正、负离子模式下采集质谱数据。根据多级质谱提供的精确分子量和碎片离子信息,并与对照品和文献报道进行比较,在不同栽培品种白芍的正、负离子模式下共鉴定出36种相同成分。在负离子模式下,两组样品分离良好;具体筛选并鉴定出17种含量有显著差异的成分,获得了“亳白芍”中独特的1种成分。采用高效液相色谱(HPLC)在Agilent HC-C₁₈(4.6 mm×250 mm,5 μm)色谱柱上进行定量分析,以0.1%磷酸水溶液(A)-乙腈(B)为流动相进行梯度洗脱,流速为1.0 mL·min⁻¹。柱温为30℃,检测波长为230 nm。建立了一种同时测定不同栽培品种白芍中8种活性成分(没食子酸、氧化芍药苷、儿茶素、芍药苷、芍药内酯苷、没食子酰芍药苷、1,2,3,4,6-O-五没食子酰葡萄糖、苯甲酰芍药苷)的HPLC方法。在所考察的线性范围内获得了良好的线性关系,相关系数(r>0.999 0)良好,方法学考察表明该方法具有良好的精密度、重复性和稳定性。平均回收率为90.61%~101.7%,相对标准偏差为0.12%~3.6%(n = 6)。UPLC-Q-TOF-MS为白芍化学成分的鉴定提供了一种快速有效的定性分析方法,所建立的HPLC方法简便、快速、准确,可为不同栽培品种白芍的种质资源评价和药材质量评价提供科学依据。