title: Integrated standardization concept for Angelica botanicals using quantitative NMR.
abstract: Despite numerous in vitro/vivo and phytochemical studies, the active constituents of Angelica sinensis (AS) have not been conclusively identified for the standardization to bioactive markers. Phytochemical analyses of AS extracts and fractions that demonstrate activity in a panel of in vitro bioassays, have repeatedly pointed to ligustilide as being (associated with) the active principle(s). Due to the chemical instability of ligustilide and related issues in GC/LC analyses, new methods capable of quantifying ligustilide in mixtures that do not rely on an identical reference standard are in high demand. This study demonstrates how NMR can satisfy the requirement for simultaneous, multi-target quantification and qualitative identification. First, the AS activity was concentrated into a single fraction by RP-solid-phase extraction, as confirmed by an alkaline phosphatase, (anti-)estrogenicity and cytotoxicity assay. Next, a quantitative (1)H NMR (qHNMR) method was established and validated using standard compounds and comparing processing methods. Subsequent 1D/2D NMR and qHNMR analysis led to the identification and quantification of ligustilide and other minor components in the active fraction, and to the development of quality criteria for authentic AS preparations. The absolute and relative quantities of ligustilide, six minor alkyl phthalides, and groups of phenylpropanoids, polyynes, and poly-unsaturated fatty acids were measured by a combination of qHNMR and 2D COSY. The qNMR approach enables multi-target quality control of the bioactive fraction, and enables the integrated biological and chemical standardization of AS botanicals. This methodology can potentially be transferred to other botanicals with active principles that act synergistically, or that contain closely related and/or constituents, which have not been conclusively identified as the active principles.
authors: UIC/NIH Center for Botanical Dietary Supplements Research, Department of Medicinal Chemistry and Pharmacognosy and PCRPS, M/C 781, College of Pharmacy, University of Illinois at Chicago, Chicago, IL 60612, USA.
authors: Tanja
authors: T
authors: Gödecke
authors: Ping
authors: P
authors: Yao
authors: José G
authors: JG
authors: Napolitano
authors: Dejan
authors: D
authors: Nikolić
authors: Birgit M
authors: BM
authors: Dietz
authors: Judy L
authors: JL
authors: Bolton
authors: Richard B
authors: RB
authors: van Breemen
authors: Norman R
authors: NR
authors: Farnsworth
authors: Shao-Nong
authors: SN
authors: Chen
authors: David C
authors: DC
authors: Lankin
authors: Guido F
authors: GF
authors: Pauli
mesh_terms: D029969:Angelica / Q000737:chemistry*; D000972:Antineoplastic Agents, Phytogenic / Q000737:chemistry / Q000494:pharmacology; D015415:Biomarkers; D002460:Cell Line; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D018517:Plant Roots / Q000737:chemistry; D015203:Reproducibility of Results
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D015415:Biomarkers