RESUMO
ObjectiveTo investigate the effect of microemulsion on the distribution of index components in different phases of Zexietang extract based on high performance liquid chromatography(HPLC) and phase separation process. MethodParticle size meter and transmission electron microscope were used to characterize the colloidal particles in blank microemulsion, aqueous extract of Zexietang and microemulsion extract of Zexietang. The phase separation process was established by high-speed centrifugation and dialysis, and based on this process, the aqueous extract and microemulsion extract of Zexietang were separated into the true solution phase, the colloidal phase and the precipitation phase, respectively. The contents of six components, including atractylenolide Ⅲ, atractylenolide Ⅱ, 23-acetyl alisol C, alisol A, alisol B and alisol B 23-acetate, were determined by HPLC with the mobile phase of water(A)-acetonitrile(B) for gradient elution(0-5 min, 40%-43%B; 5-20 min, 43%-45%B; 20-45 min. 45%-60%B; 45-75 min, 60%-80%B). The solubility of the index components in water and microemulsion was determined by saturation solubility method. ResultThe colloidal particles in the aqueous extract, microemulsion extract and blank microemulsion were all spherical, and the particle size, polydispersity index(PDI) and Zeta potential of the colloidal particles were in the order of aqueous extract >microemulsion extract >blank microemulsion. The results of phase separation showed that the colloidal phase and the true solution phase could be completely separated by dialysis for 2.5 h, and the phase separation process was tested to be stable and feasible. Compared with the aqueous extract of Zexietang, the use of microemulsion as an extraction solvent could increase the contents of atractylenolide Ⅲ, 23-acetyl alisol C, atractylenolide Ⅱ , alisol A, alisol B and alisol B 23-acetate by 3.75, 6.82, 35.47, 10.66, 35.41, 27.75-fold, and could increase the extraction efficiencies of the latter five constituents by 2.03, 1.15, 1.70, 6.43, 5.53 times. The solubility test showed that the microemulsion could significantly improve the solubility of atractylenolide Ⅱ, alisol A, alisol B and alisol B 23-acetate, but it had less effect on the solubility of atractylenolide Ⅲ and 23-acetyl alisol C. ConclusionMicroemulsion can improve the extraction efficiency and increase the distribution of the index components in the colloidal phase state of Zexietang to different degrees, providing a reference for the feasibility of microemulsion as an extraction solvent for traditional Chinese medicine.
RESUMO
ObjectiveBased on serum pharmacochemistry and ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) the transitional components in the serum of rats after intragastric administration of water extract of Alismatis Rhizoma(AR)and salt-processed Alismatis Rhizoma(SAR) were compared. MethodSD rats were randomly divided into blank group, AR group(10 g·kg-1) and SAR group(10 g·kg-1), 3 rats in each group, the administration groups were given AR and SAR aqueous extracts by gavage, respectively, and the blank group was given an equal volume of drinking water by gavage once in the morning and once in the evening, for 3 consecutive days. Sixty minutes after the last administration, blood was collected from the eye orbits, and the serum samples were prepared. The serum samples were prepared on an ACQUITY UPLC BEH C18 column(2.1 mm×50 mm, 1.7 μm) with the mobile phase of acetonitrile(A)-0.1% formic acid aqueous solution(B) in a gradient elution(0-10 min, 10%-50% A; 10-27 min, 50%-95%A; 27-27.1 min, 95%-10% A; 27.1-30 min, 10%A), the data were collected at a flow rate of 0.3 mL·min-1 in positive ion mode with a scanning range of m/z 100-1 200. Based on the self-constructed chemical composition library of AR, the total ion flow diagrams and secondary MS fragmentation information of the aqueous extracts of AR and SAR, as well as the administered serum and the blank serum, were compared with each other by UNIFI 1.9.2, so as to deduce the possible blood-migrating constituents and their cleavage patterns in the aqueous extracts, and the response intensity ratios of each chemical component were calculated before and after processing. ResultA total of 20 components, including 5 prototypical components and 15 metabolites, were analyzed and deduced from the serum of rats given aqueous extract of AR. And 14 components, including 5 prototypical components and 9 metabolites, were analyzed and deduced from the serum of rats given aqueous extract of SAR. Of these, 13 components were common to both of them, including 5 prototypical components and 8 metabolites. The 5 prototypical components were 16-oxoalisol A, alisol A 24-acetate, alisol A, alisol B and alisol C. The metabolites were mainly involved in phase Ⅰ metabolism(oxidation) and phase Ⅱ metabolism(glucuronidation). There was a big change in the intensity of response of the common components before and after salt-processing, and the response intensities of the prototypical components, 16-oxoalisol A, alisol B and alisol C, were elevated, while the type and response intensity of metabolites were generally decreased, and it was hypothesized that the metabolic rate of terpenoids might be slowed down after salt-processing of AR, so that the blood-migrating constituents could participate in the metabolism of the body more in the form of prototypes. ConclusionSalt-processing of AR may promote the absorption of prototypical components into the blood by slowing down the metabolic rate of terpenoids, which can provide support for the research on material basis of AR and SAR.