[Identification of chemical constituents of Shuangshen Fuzheng Powder and its tissue distribution characteristics in rats by UHPLC-Q-Exactive-Orbitrap-HRMS].

Abstract

Shuangshen Fuzheng Powder has been clinically used for the prevention and treatment of lung cancer for many years, demonstrating favorable efficacy, yet its pharmacodynamic material basis remains to be further investigated. This study employed ultra-high-performance liquid chromatography coupled with quadrupole-electrostatic field Orbitrap high-resolution mass spectrometry(UHPLC-Q-Exactive-Orbitrap-HRMS) to rapidly identify the chemical components of Shuangshen Fuzheng Powder and to investigate the distribution of major prototype components in blood and tissue after intragastric administration in rats. An ACQUITY UPLC BEH C_(18) column(2.1 mm×100 mm, 1.7 μm) was used, with 0.1% formic acid in water(A) and methanol(B) as the mobile phase under gradient elution. Mass spectra were acquired in positive and negative ion full-scan/data-dependent secondary scan mode(Full MS/dd-MS~2). Elemental compositions were fitted using Xcalibur 4.2 software based on the relative retention time of ion peaks and primary fragment ion information, and component identification was performed by comparison with reference standards, literature, and databases using secondary fragment ion information. This approach enabled the identification of both chemical constituents of Shuangshen Fuzheng Powder and the prototype components present in blood and various tissue. A total of 130 chemical components were identified, including 78 terpenoids(saponins), 16 flavonoids, 13 nucleotides, 6 amino acids, 5 fatty acids, 4 organic acids, and 8 other compounds. After intragastric administration to rats, 20, 15, 46, 66, 46, 61, 38, and 40 prototype components were detected in serum, brain, lung, heart, liver, spleen, kidney, and ileum, respectively. Thirteen components, such as naringenin, ginsenoside R_1, ginsenoside Rg_5, ginsenoside Re, ginsenoside Rg_1, ginsenoside F_2, ginsenoside Rh_1, ginsenoside Ro, ginsenoside Rb_1, notoginsenoside R_1 and notoginsenoside R_2, yesanchinoside B, and malonyl ginsenoside Rb_1, were detected both in serum and across all tissue. This study demonstrates that high-resolution mass spectrometry allows rapid identification of the chemical constituents of Shuangshen Fuzheng Powder and the distribution of its prototype components in blood and tissue. These findings provide a reference for elucidating the pharmacodynamic material basis of Shuangshen Fuzheng Powder and for conducting pharmacokinetic studies of its active components in different tissue.