Identification of metabolites of Danggui Buxue Tang in rat urine by liquid chromatography coupled with electrospray ionization time‐of‐flight mass spectrometry

Abstract

A method coupling liquid chromatography with electrospray ionization time-of-flight mass spectrometry (LC/ESI-TOF/MS) has been developed for rapid and sensitive analysis of rat urinary metabolite profile of Danggui Buxue Tang (DBT), a well-known Chinese herbal formula. After oral administration of DBT, urine samples were collected during 0-24 h, and then pretreated by solid-phase extraction. A total of 68 compounds including 13 parent compounds and 55 metabolites were detected in the drug-containing urines compared with blank urines. The total analytical time was less than 20 min. Metabolites of DBT were identified using dynamic adjustment of the fragmentor voltage to produce structure-relevant fragment ions. By using this approach, the mass accuracy of precursor and fragment ions was typically within +/-5 ppm of the theoretical values, and enabled the identification of 43 metabolites including 27 isoflavanoid and 16 phthalide metabolites. Our results indicated that glucuronidation and sulfation were the major metabolic pathways of isoflavonoids, while glutathione conjugation, glucuronidation and sulfation were the main metabolic pathways of phthalides. No saponin-related metabolites were detected. The results of the present study provided important structural information relating to the metabolism of DBT. Furthermore, this work demonstrated the potential of the LC/ESI-TOF/MS approach for identification of metabolites from Chinese herbal medicines in urine.

Identification of the absorbed components and metabolites in rat plasma after oral administration of Rhizoma Chuanxiong decoction by HPLC–ESI-MS/MS

“…Similarly, M1, M3, M5, M9 were identified as senkyunolide J-related metabolites owing to the presence of diagnostic fragment ions related to senkyunolide J, such as 209, 191, 163, 153. While M8, M12 were designated as butylidenephthalide-related metabolites due to the shifts of exact masses, characteristic neutral losses and some diagnostic ions related to butylidenephthalide [28]. glutathione conjugate of senkyunolide J. ESI-MS/MS spectra of M5 and M6 at positive ion mode are presented in Fig.…”

Correlative analysis of metabolite profiling of Danggui Buxue Tang in rat biological fluids by rapid resolution LC–TOF/MS

“…1 shows the typical total ion chromatograms (TICs) of bile and plasma samples in the positive ion mode respectively. By comparing with the blank and drug-containing biological fluids, more endogenous interferences were observed in bile and plasma than in urine [19].…”

“…Other reagents and chemicals were of analytical grade. DBT oral solution and mixture of authentic standards were prepared by our laboratories as described in the previous study [19].…”

“…As for metabolite identification, TOFMS enables not only the fast detection of data for all expected and unexpected metabolites from a single run, but also reliable and accurate identification for molecular formula and related biotransformation of metabolites [16][17][18]. Previously, we have developed an RRLC-TOFMS method for characterization of rat urinary metabolite profile of DBT [19], which provided important structural information relating to the metabolism of DBT. As an ongoing work, the aims of this study are to further describe the metabolite profiles of DBT in rat bile and plasma.…”

HPLC method with fluorescence detection for the determination of ligustilide in rat plasma and its pharmacokinetics

“…To date, there have been some reports on the quantitative analysis of LIG in different plant materials or their extracts by gas chromatography (GC)-flame ionization detection (FID) (Fang et al, 1979;Li et al, 2001), capillary zone electrophoresis (CZE) (Ji et al, 1999), gas chromatography-mass spectrometry (GC-MS) Cho et al, 2007;Deng et al, 2005;Hu & Ding, 2006;Kim et al, 2006a,b;Lao et al, 2004;Li et al, 2006a), liquid chromatography-mass spectrometry (LC-MS) (Lin et al, 1998;Liu et al, 2009;Lu et al, 2004;Qi et al, 2008;Yi et al, 2007;Zschocke et al, 1998), liquid chromatography (LC) (Chao & Chao, 2004;Cui et al, 2006;Hu et al, 2005a,b;Li et al, 2006b;Lu et al, 2005aLu et al, ,b, 2009), bioactivity-guided fractionation method combining countercurrent chromatography (CCC), the MTS cell viability assay and gas chromatography (GC) (Yeh et al, 2012). Although few methods have been reported for the quantification of LIG in biosamples Guo et al, 2009Guo et al, , 2011Li et al, 2009Li et al, , 2011Shi et al, 2006;Yan et al, 2008;Zuo et al, 2011), which are commonly measured as biomarkers. As for extraction of LIG from biological samples, the analytical methods involved multiple steps of liquid-liquid extraction, which used relatively toxic agents such as n-hexaneether, followed by evaporation to prepare and concentrate the sample prior to injection to an LC system, the pretreatment of the biological samples were laborious and time-consuming.…”