An effective integrated method for comprehensive identification of eighty-five compounds in Zhi-Zi-Da-Huang decoction by HPLC-DAD-ESI-MS (TOF) and HPLC-DAD-ESI-MS/MS (QqQ) without the help of reference standards

Abstract

decoction (ZZDHD) is an effective formula to treat alcoholic liver disease in China for nearly 2000 years. However, compounds in the formula are still not clear due to the complexity of the coexisting constituents. In this study, a sensitive integrated method based on HPLC-DAD-ESI-MS (TOF) and HPLC-DAD-ESI-MS/MS (QqQ) was developed and 85 chemical compounds in ZZDHD were identified. By TOF/MS, the quasi-molecular ions of the constituents were deduced according to the accurate mass of corresponding positive and negative ions, and product ion scan was then carried out using QqQ/MS. The fragments of each compound were observed by TOF/MS and/or QqQ/MS. Compounds with different chemical structures and concentration levels were tentatively identified according to their chromatographic retention time, UV absorption and MS data. Fragmentation patterns of six types were proposed to verify the identification results. It was found that at TOF/MS mode, a retro diels-alder (RDA) reaction occurred at bonds 1 and 3 in C-ring of flavanone 7-O-diglycosides when 4 0 -OH exists at B-ring. While the reaction was not observed if a methoxyl group appeared at the same site of flavanone skeleton. Another important phenomenon was noticed that 3-sinapoyl-5-caffeoylquinic acid, 4-sinapoyl-3-caffeoylquinic acid, and sinapylglucoside were detected only in ZZDHD while they were not found in four component herb extracts. The phenomenon revealed that changes of compounds happened when the component herbs were decocted together. The results in the paper are favorable of the clinic dosage and have laid a foundation for the subsequent research of ZZDHD in vivo.

Chemical profiling approach to evaluate the influence of traditional and simplified decoction methods on the holistic quality of Da-Huang-Xiao-Shi decoction using high-performance liquid chromatography coupled with diode-array detection and time-of-flight

“…Monoterpenoids are the main compounds from the crude drug ZZ. Nine monoterpenoids were tentatively identified in this study [26,27]. Peaks 46 and 47 were isomers and formed the similar adduct ions.…”

“…Based on these fragmentation ions and retention times, peaks 12 and 13 were tentatively identified as caryoptosidic acid and shanzhiside, respectively [25]. Peaks 14,15,19,20,34,67,68, and 74 were identified similarly based on the fragmentation pathways as well as the literature reports [26] and the assignment of these compounds is summarized in Supporting Information Table S1.…”

An untargeted metabolomics-driven approach based on LC–TOF/MS and LC–MS/MS for the screening of xenobiotics and metabolites of Zhi-Zi-Da-Huang decoction in rat plasma

“…According to the previous studies on the chemical and structure information of ZZDHD [3], iridoid glycosides, monoterpenoids, flavones and their glycosides, anthraquinones are four main types of components. As most of the metabolites remained the skeleton structural features of the main prototype constituents, the analysis of the fragmentation pathways of prototype constituents greatly facilitated the characterization of metabolites from ZZDHD.…”

“…According to the previous publication [3], the typical fragmentation rules for iridoid glycosides contained the retro diels-alder (RDA) reaction of iridoid skeleton and loss of a glucosyl moiety (162 Da). In this study, based on the untargeted metabolomics-driven approach and the elemental composition, fragmentation behaviors of the metabolites and expected neutral losses by biotransformation such as H 2 O (−18 Da) or glucuronyl (−176 Da), a total of ten iridoid-related metabolites were tentatively identified in vivo.…”

“…Modern pharmacological evaluations have demonstrated that ZZDHD exhibits promising hepatoprotective effect [1]. Although a total of 85 chemical components in ZZDHD were identified or tentatively characterized by LC-TOF/MS and LC-QqQ/MS [2,3], the active ingredients of ZZDHD which is crucial for further investigation of therapeutic effect of the formula are still not so clear. One feasible way is to clarify "what are absorbed" and "what are produced" of ZZDHD in vivo since only the xenobiotics and metabolites detected here could responsible for the therapeutic effects [4][5][6], in most cases.…”