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Application of a strategy based on metabolomics guided promoting blood circulation bioactivity compounds screening of vinegar-Institution of Basic Theory-China Academy of Chinese Medical Sciences- China

Abstract

Zhenli Liu

Vinegar has been adopted as favoring dating from around 3000 BC in Asian, European and other traditional cuisines of the world .As evidences accumulated, vinegar was proved to exhibit therapeutic properties, including blood pressure reduction , antioxidant activity , antibacterial activity , reduction in the effects of diabetes and prevention of cardiovascular disease . It is also used as a kind of accessory documented in Lei’s treatise on processing of drugs (LeigongpaozhiLun) (618–907 AD). Numerous Chinese medicines such as Frankincense, Rhizoma Corydalis were believed to enhance the promoting blood circulation therapeutic efficiency after preparation by vinegar . Fruitful researches have been carried on the herbal enhancement of therapeutic efficiency after processing ,but there are few relative reports concerning the blood circulation bioactivity of vinegar Vinegar accumulate an overwhelming variety of metabolites that play nondeductible roles in health benefit. During recent years, many studies employed GC–MS technique for quality control and determination of vinegar. Alcohols, organic acids, amino acids, carbohydrates, esters and various micro-constituents were proved to present in vinegar [8]. The previous results showed that the contents of most conventional ingredients (organic acids, free amino acids, carbohydrates) were increased during aging process. Tetramethylpyrazine (TMPZ), a kind of alkaloid metabolites yielding during aging process of vinegar, was used in clinical trials since the 1970s [9]. Reports indicate that TMPZ reduces arterial resistance  and increases coronary and cerebral blood flow . A number of alkaloid metabolites are developed as clinical drugs found to have significant biological activities (e.g  berberine and paclitaxel) . Hence variation of alkaloid metabolites should not be overlooked for their exhibit notable function properties.

Rice vinegar (RV) and white vinegar (WV) as daily favoring, have also used as accessory in traditional Chinese medicine processing. As we know, the promoting blood circulation efficiency could be enhanced when herbs processed by vinegar. Number of reports focused on health benefits derived by consumption of vinegar. However, few concerned the blood circulation bioactivity.

 In this paper, a metabolomics guided strategy was proposed to elaborate on the chemical constituents’ variation of two kinds of vinegar. GC–MS coupled with multivariate statistical analysis were conducted to analyze the chemical components in RV and WV and discriminate these two kinds of vinegar. The anti-platelet activities in vitro were investigated by whole blood aggregometry platelet test. And the anticoagulant activities were monitored by the whole blood viscosity, plasma viscosity, packed cell volume, prothrombin time, and four coagulation tests (PT, TT, APTT, FIB) in vivo .Results were found by keen observation of GC–MS conditions Chromatographic parameters such as column type, carrier gas flow, temperature rate, and ion source temperature were adjusted to be able to obtain the best separation for the compounds. The Rxi-50 capillary column obtained the best separation. The carrier gas at flow rate of 1.0 mL/min and the 250 °C ion source temperature were proved to be the most suitable. Established chromatographic conditions and mass spectra conditions are listed in “Gas chromatography–mass spectrometry analysis’’. Metabolic profiles of RV and WV Five batches of WV and RV with aging time of 1, 4, 5, 7, 14, 20, 30  months were analyzed. Representative GC– MS fingerprints are presented in Fig.  1. A total of 53 compounds were detected, including different kinds of alcohol, organic acids, amino acids, aldehydes, phenols, ketones, heterocyclic, which were same as those reported in literatures [1, 20]. PCA and OPLS-DA were utilized to classify the metabolic phenotypes and identify the differentiating metabolites. A PCA score plot for first and second principal components was utilized to depict the general variation among the samples of two dosage forms (R2 X  =  0.78, Q2 =  0.987).

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