Li Nanxin, Zhang Jinju, Zhang Ying, Ma Zhiguo, Liao Jinhong, Cao Hui, Wu Menghua
1College of Pharmacy, Lingnan Traditional Chinese Medicine Research Center of Jinan University, Lingnan Branch of National Engineering Research Center for Modernization of Traditional Chinese Medicine, Jinan University, Guangzhou, 510632 Guangdong China.
2Guangdong Provincial Key Laboratory of Research on Pharmacodynamic Substances and Innovative Drugs of Traditional Chinese Medicine, Guangzhou, 510632 Guangdong China.
Chin Med. 2020 Feb 18;15:19. doi: 10.1186/s13020-020-00301-5. eCollection 2020.
Danmu preparations (Danmu Capsule and Danmu Syrup), which are made from stem extracts, have good clinical efficacy in acute tonsillitis, acute pharyngitis and upper respiratory tract infection. However, there is currently no reliable and systematic method to control the quality of these two Danmu preparations.
Using high-performance liquid chromatography (HPLC) coupled with diode array detection (DAD), the fingerprints of the Danmu preparations were established at 250 nm to comprehensively investigate the stability of preparation process. The chemical constituents in the Danmu preparations were separated and identified by HPLC coupled with quadrupole-time-of-flight high-definition mass spectrometry (HPLC-Q-TOF-MS). And seven major components were simultaneously determined at dual wavelengths (250 nm, 326 nm).
The results of HPLC fingerprint similarity evaluation showed that the similarity values of 25 batches of Danmu preparations were more than 0.993. Twenty-three compounds, including 10 alkaloids, 6 phenolic acids, 2 iridoids, and 5 unknown compounds, were identified or tentatively characterized according to the retention times and MS/MS fragment patterns of compounds. The developed assay method of seven components was validated with acceptable linearity, precision, repeatability, stability and recovery. The contents of strictosamide belonging to alkaloids as the most abundant constituent in Danmu Capsule and Danmu Syrup were 43,681.20-99,652.49 μg/g and 1567.83-2427.25 μg/mL respectively. The contents of protocatechuic acid which were the highest in measured phenolic acids were 2633.01-7739.78 μg/g in Danmu Capsule and 192.05-448.71 μg/mL in Danmu Syrup, respectively. As an iridoid, the contents of sweroside in Danmu Capsule and Danmu Syrup were 1573.82-2789.81 μg/g and 70.32-182.81 μg/mL, respectively.
The established qualitative analysis method of fingerprint can be used to attain standardization, uniformity and stability of the preparation process. Meanwhile, the quantitative analysis in this study can be used as an accurate assay method for preparations.
胆木制剂（胆木胶囊和胆木糖浆）由茎提取物制成，在急性扁桃体炎、急性咽炎及上呼吸道感染方面具有良好的临床疗效。然而，目前尚无可靠且系统的方法来控制这两种胆木制剂的质量。
采用高效液相色谱（HPLC）结合二极管阵列检测（DAD），在250nm波长下建立胆木制剂的指纹图谱，以全面考察制剂过程的稳定性。通过HPLC结合四极杆-飞行时间高清质谱（HPLC-Q-TOF-MS）对胆木制剂中的化学成分进行分离和鉴定。并在双波长（250nm、326nm）下同时测定七种主要成分。
HPLC指纹图谱相似度评价结果显示，25批次胆木制剂的相似度值均大于0.993。根据化合物的保留时间和MS/MS碎片模式，鉴定或初步表征了23种化合物，包括10种生物碱、6种酚酸、2种环烯醚萜和5种未知化合物。所建立的七种成分测定方法经方法学验证，具有良好的线性、精密度、重复性、稳定性和回收率。作为生物碱中含量最丰富的成分，胆木胶囊和胆木糖浆中马钱苷的含量分别为43681.20 - 99652.49μg/g和1567.83 - 2427.25μg/mL。在所测定的酚酸中含量最高的原儿茶酸，在胆木胶囊中的含量为2633.01 - 7739.78μg/g，在胆木糖浆中的含量为192.05 - 448.71μg/mL。作为环烯醚萜，胆木胶囊和胆木糖浆中獐牙菜苷的含量分别为157,3.82 - 2789.81μg/g和70.32 - 182.81μg/mL。
所建立的指纹图谱定性分析方法可用于实现制剂过程的标准化、一致性和稳定性。同时，本研究中的定量分析可作为制剂的准确测定方法。
