The Efficient Synthesis and Anti-Fatigue Activity Evaluation of Macamides: The Unique Bioactive Compounds in Maca.

Macamides are a class of amide alkaloids that are only found in maca and are widely considered to be its bioactive marker compounds. More than thirty macamide monomers have been identified in recent years; however, it is difficult to obtain a single macamide monomer from the maca plant because of their similar structures and characteristics. We used the carbodiimide condensation method (CCM) to efficiently synthesize five typical macamides, including N-benzyl-hexadecanamide (NBH), N-benzyl-9Z,12Z,15Z-octadecenamide, N-(3-methoxybenzyl)-9Z,12Z-octadecenamide, N-benzyl-9Z,12Z-octadecenamide, and N-(3-methoxybenzyl)-9Z,12Z,15Z-octadecadienamide. All the synthesized macamides were purified by a one-step HPLC with a purity of more than 95%. NBH is the most abundant macamide monomer in natural maca, and it was selected to evaluate the anti-fatigue effects of macamides. The results indicated that NBH could enhance the endurance capacity of mice by increasing liver glycogen levels and decreasing blood urea nitrogen, lactate dehydrogenase, blood ammonia, and blood lactic acid levels. Macamides might be the active substances that give maca its anti-fatigue active function.

Thrombin-based discovery strategy of bioactive-chemical quality marker combination for pollen of Typha orientalis by metabolomics coupled with chemometrics.

BACKGROUND: It is of utmost significance to choose the bioactive components as quality markers for ensuring the effectiveness of traditional Chinese medicine (TCM). Nonetheless, some markers are able to assess effectively the quality of TCM without considering the pharmacological mechanisms and intrinsic chemical complexities. OBJECTIVE: This underscores the need to discover new and efficient markers which can assess both quality and mechanism of action. Herein, a strategy of bioactive-chemical quality marker combination was proposed to improve the level of the quality control of TCM by metabolomics coupled with chemometrics. METHODS: A four-step plan was followed. Firstly, acquisition of metabolic features and component characterization of different batches of pollen of Typha orientalis C.Presl were performed using UHPLC-Q-TOF/MS. Secondly, the direct inhibitory effects of pollen of T. orientalis on thrombin was assessed by using chromogenic substrate method together with HPLC. Thereafter, bioactive-chemical marker combination associated with anti-thrombin segregation was screened using supervised classifiers. Finally, quantitative assay and prediction-model of selected markers were established for guarantying the quality of pollen of T. orientalis. RESULTS: A total of 22 compounds were annotated based on comparison with previous work from pollen of T. orientalis by UHPLC-Q-TOF/MS. Citric acid and linolenic acid inhibited the thrombin activity with IC50 values, 0.52 ± 0.02 and 0.51 ± 0.02 mg/mL, respectively. A bioactive-chemical marker combination including citric acid, linolenic acid, typhaneoside, and isorhamnetin-3-O-neohesperidoside were discovered and selected as quality markers for evaluation of pollen of T. orientalis according to their capacity for inhibiting thrombin. CONCLUSION: The thrombin-based discovery strategy of bioactive-chemical marker combination was a powerful tool for screening the quality markers for evaluation of pollen of T. orientalis.

Method establishment for upgrading chemical markers in pharmacopoeia to bioactive markers for biological standardization of traditional Chinese medicine.

Quality surveillance on authentication, safety and efficacy of proprietary Chinese medicines (pCm) are certainly the top priorities for the industries. Nowadays, the quality control system adopted is mainly chemical marker-oriented, concerning basically the correct use of raw material and safety issues, while the biological activities of the chemical marker(s) are seldom considered. Hence, there is an undefined relationship between the amount of chemical markers and the claimed pharmacological activities. In view of the need in identifying appropriate markers for biological standardization of pCm products, the present study aimed to establish a systematic methodology for verifying whether the chemical marker of a traditional Chinese medicine (TCM) listed in Chinese Pharmacopoeia could be upgraded to a bioactive marker with certain efficacy in treating a particular disease. Our proposed methodology included a series of work on extraction, quantification, literature search and in vivo pharmacological experiments, in which the water extractability, biological effects at theoretical dose and oral bioavailability of the candidate chemical markers were all taken into consideration. The feasibility and implication of this bioactive markers verification methodology were further elaborated. Our findings will serve as the foundation for further research and development of biological standardization of TCM.

Esculetin as bioactive marker: towards a rational scientific approach for the treatment of hyperuricemia using Traditional Chinese Medicine

Traditional Chinese Herbal Medicine has been used to prevent and cure disease in China for thousands of years and has gained global interest in recent decades. The Erding formula is a Chinese Pharmacopeia (ChP)-listed herbal preparation used for treating sore throat, carbuncles and boils. Esculetin is a ChP quality control (QC) marker for these indications. A previous study found that a new indication, hyperuricemia, can be added to the Erding formula. Therefore, this study aimed to evaluate whether the traditionally used marker, esculetin, still has bioactivity for hyperuricemia, which is substantially different from the original indications. The study analyzed the quantity of esculetin by high-performance liquid chromatography, assessed the therapeutic effect of esculetin using animal model, and then characterized esculetin and its metabolites in serum via ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry. The results showed that the esculetin content in the aqueous Erding extract was 0.26±0.05% (w/w). Both the Erding extract and esculetin significantly reduced uric acid levels. Six metabolites of esculetin were identified in mice serum. This study revealed a rational scientific approach to prove esculetin is a reliable bioactive and QC marker for Erding formula in hyperuricemia treatment which contributed to ensure product quality and therapeutic efficacy.

Validated high performance thin layer chromatography method for the quantification of bioactive marker compounds in Draksharishta, an ayurvedic polyherbal formulation

ABSTRACT Draksharishta is an ayurvedic polyherbal formulation with Draksha (Vitis vinifera L., Vitaceae) as chief ingredient prescribed for digestive impairment, respiratory disorders and weakness. These herbal medicines containing biologically active compounds play a significant role. Therefore it is necessary to carry out the chemical standardization of bioactive marker compounds present in the polyherbal ayurvedic formulation like Draksharishta. The aim of the present work was to develop and validate a HPTLC method for determination of gallic acid, catechin and resveratrol in commercially available marketed and in-house prepared formulations of Draksharishta. This is the first report of quantification of bioactive marker compound resveratrol using HPTLC in Draksharishta. The method employed silica gel precoated thin layer chromatography plates with F254 as the stationary phase. The respective mobile phases were used to develop the plates which separated bands according to the marker compound. Camag scanner V was used for densitometric scanning. Further, the method was validated according to International Conference of Harmonization (ICH) guidelines. The Rƒ values of the three marker compounds were measured. Correlation coefficients were calculated from the standard graph of linearity. Accuracy, precision and recovery were all within the required limits. The developed HPTLC methods for bioactive marker compounds present in in-house and marketed formulations were found to be simple, accurate, precise and robust.