UHPLC coupled with mass spectrometry and chemometric analysis of Kang‐Ai injection based on the chemical characterization, simultaneous quantification, and relative quantification of 47 herbal alkaloids and saponins

Abstract

Kang‐Ai injection, which is composed of Astragali Radix, Ginseng Radix et Rhizoma, and kushenin, is extensively used in China as an adjuvant therapy for many types of cancer and chronic hepatitis B. In the present study, 47 herbal compounds (11 alkaloids, 8 astragalosides, and 28 ginsenosides), were detected in Kang‐Ai injection by ultra‐high‐performance liquid chromatography coupled to quadrupole time‐of‐flight tandem mass spectrometry, of which 31 were identified using authentic standards. Additionally, a practical ultra‐high‐performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry method was employed for simultaneous quantitative detection (31 available compounds), and relative quantitative detection (16 unavailable compounds) within 10 min. The limit of detection and limit of quantification was 0.11‐2.22 and 0.53‐11.08 ng/mL, respectively. Altogether, content levels of each compound ranged from 0.03 to 9835.57 μg/mL. Furthermore, chemometric analysis indicated oxymatrine, astragaloside IV, ginsenosides Rg1 and Re, and matrine had the greatest effect on concentration fluctuation. Therefore, we suggested these five compounds should be monitored during the manufacturing process. This method can be applied to provide crucial chemical profiles and quality assessments for Kang‐Ai injection, guaranteeing the safety, effectiveness, and controllability of the drug in clinics.

Multiple circulating alkaloids and saponins from intravenous Kang-Ai injection inhibit human cytochrome P450 and UDP-glucuronosyltransferase isozymes: potential drug-drug interactions

“…3A), which could be mainly attributed to the contribution of oxymatrine. This is because the content level of oxymatrine in Kang-Ai injection (37731.41 μM) is far higher than those of other herbal compounds (0.23~158.95 μM) (Additional le 1: Table S1) [9]. Furthermore, it was notable that the deglycosylation of astragaloside IV (B2) to sapogenin-cycloastragenol strongly increased the inhibitory effects towards UGT1A8 (K i = 0.034 µM) and 2B7 (K i = 20.98 µM) [15].…”

“…S3A & Table S1). Based on their individual content level from Kang-Ai injection [9], these compounds were graded into ve dose levels (Additional le 1: Fig. S3B & Table S1): Level I was over 1000 μg/mL for only oxymatrine (A3); Level II was between 100 and 1000 μg/mL for ginsenosides Rg1 (C6) and Re (C9); Level III was ranged from 10 to 100 μg/mL for matrine (A1), astragaloside IV (B2), notoginsenoside R2 (C3), ginsenoside Rg2 (C5), Rf (C7) and Rc (D3); Level IV was from 1 to 10 μg/mL for astragaloside III (B3), VI (B4), V (B5), notoginsenoside R1 (C8), ginsenoside Rh1 (C1), Rd (D2), Rb2 (D4), Rb1 (D5), Rh4 (E2), Rg6 (E3), F4 (E4); Level V was all less than 1 μg/mL for oxysophocarpine (A2), isoastragaloside IV (B1), ginsenoside F1 (C2), F3 (C4), F2 (D1), Rk3 (E1).…”

“…It consists of Astragali Radix (Huangqi), Ginseng Radix et Rhizoma (Renshen) and kushenin (Kushensu), and is indicated for many cancers, leukopenia and chronic hepatitis B [6][7][8]. Previously, we have nished the chemical identi cation and content determination of the main alkaloids (oxymatrine, matrine), astragalosides (astragaloside IV) and ginseng saponins (ginsenosides Rg1, Re) in Kang-Ai injection [9]. Despite the increasing understanding of Kang-Ai injection in chemical and pharmacological studies, the systemic exposure and excretion of its bioactive compounds after administration remain to be elucidated.…”

Identification of bilobetin metabolites, in vivo and in vitro, based on an efficient ultra‐high‐performance liquid chromatography coupled with quadrupole time‐of‐flight mass spectrometry strategy

“…Owing to its many advantages, ultra-high-performance liquid chromatography has been widely used in the analysis of complex components of natural products, analysis of biological samples with complex matrices, discovery of disease biomarkers, and metabolomics [14][15][16][17][18][19]. The particle size of the column used in ultra-high-performance liquid chromatography is 1.7-3.5 µm, and the liquid chromatography system can work in the pressure range of 6000-15 000 psi.…”

Multiple circulating alkaloids and saponins from intravenous Kang-Ai injection inhibit human cytochromes P450 and UDP-glucuronosyltransferase isozymes: potential drug-drug interactions

“…Previously, we have finished the chemical identification and content determination of the main alkaloids (oxymatrine, matrine), astragalosides (astragaloside IV) and ginseng saponins (ginsenoside Rg1, Re) in Kang-Ai injection [9]. Despite the increasing understanding of Kang-Ai injection in chemical and pharmacological studies, the systemic exposure and excretion of its bioactive compounds after administration remain to be elucidated.…”