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Objective: To study and compare the content of two flavonoids and two saponins in Ziziphi Spinosae Semen from seven major producing areas in China, and to provide high quality sources of medicinal materials. Methods: Investigation and sample collection of seven major production areas were carried out in 2016 and 2017 and a total of 74 samples were collected. UPLC-UV-ELSD liquid phase method was used to determine the content of two flavones, spironol and 6’’’-ferulinyl spironol, as well as two saponins jujuboside A and jujuboside B. Results: In 2016 and 2017, the content of spinolin in the samples was 0.052%-0.102% and 0.049%-0.144%, respectively. The content of 6’’’-ferulinyl spironol was 0.021%-0.072% and 0.026%-0.088%, respectively. The content of jujuboside A was 0.016%-0.061% and 0.033%-0.054%, respectively. The content of jujuboside B was 0.008%-0.046% and 0.005%-0.046%, respectively. Conclusion: By correlation analysis, there was no significant difference in the content of each component among the seven major production areas. The producing areas with high spinosin content are Linfen, Shanxi (including Daning and Jixian), Shexian, Hebei, Qian’ an, Hebei, Qingyang, Gansu, Meixian, Shaanxi, Heyang, Shaanxi, Jianchang, Liaoning, and Jixian, Tianjin. The producing areas with high content of jujuboside A are most places of Shandong province, Neiqiu, Hebei and Weinan, Shaanxi. Considering the content of two kinds of components, Shandong Province is generally high and has little change. The correlation analysis also showed that the content of two flavonoids had good correlation, but the content of two saponins had poor correlation.
ABSTRACT
Objective: The aim of this work was to investigate the quality markers (Q-markers) of Ziziphi Spinosae Semen (ZSS) based on biotransformation by human intestinal microbiota. Methods: In this study, in vitro biotransformation of ZSS aqueous extract by normal human intestinal microbiota was analyzed using UPLC-Q-Orbitrap-MS. Furthermore, the time course of the biotransformation was studied to probe into the biotransformation mechanism of compounds in ZSS by human intestinal flora. The change rules of flavonoids, saponins and alkaloids in the incubation solution at different time points were plotted based on the percentage of peak area of compounds. Results: A total of 31 original ingredients and four metabolites were characterized in transformed ZSS aqueous extract by human intestinal microbiota. No obvious degradation was observed for benzylisoquinoline alkaloids within 24 h. As far as flavones concerned, a wide range of metabolic reactions as well as significant reaction were shown. Meanwhile, these flavonoids were completely degraded during 24 h. In addition, both jujuboside A and jujuboside B were metabolized to their saponins by deglycosylation reactions. Thus, coclaurine, zizyphusine, kaempferol-3-O-rutinoside, spinosin, vicenin II, jujuboside A, and jujuboside B were referred as prospective Q-markers. Conclusion: The results indicated that the chemical compounds in ZSS were obviously affected by transformation. Intestinal transformation studies play an important role for the elucidation of therapeutic material basis of ZSS and it should be taken into account during the process of the investigation of Q-marker.
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Objective: To establish an accelerated solvent extraction (ASE)-charged aerosol detector (CAD) method for simultaneous detection of jujuboside A, jujuboside B, spinosin, and betulinic acid in Zizyphi Spinosae Semen. Methods: The orthogonal design was applied to optimize the extraction parameters of the ASE system. The target compounds were detected by HPLC-CAD with the parameters as follow: Thermo Syncronis C18 (100 mm × 3 mm, 3 μm) column, a gradient elution program with acetonitrile-water as mobile phase at a flow rate of 0.5 mL/min, the column temperature was kept at 40℃. Detector was Corona Ultra CAD with 35℃ of nebulization temperature. Results: Optimization of the ASE parameters with orthogonal design greatly improved the extraction efficiency; All the target compounds could be simultaneously determined in a single run. Good linear relationships (0.998 3-0.999 6) and high relative recoveries were 98.46%-102.02%. Conclusion: The method is rapid, simple, accurate, and thus could be used for the quality control of Zizyphi Spinosae Semen.
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OBJECTIVE: To establish an HPLC method for simultaneous determination of ginsenoside Rg1, Re, Rb1 and jujuboside A, B in Renshen Jianpi Pellets (RSJPW). METHODS: The determination was conducted on an Eclipse XDB C18 column (4.6 mm × 250 mm, 5 μm) with the mobile phase of acetonitrile (A)-water (B) gradiently eluted at the following flow rates: 0-90 min, 1.0 mL · min-1; 90-130 min, 1.0-0.5 mL · min-1; 130-140 min, 0.5 mL · min-1. And the column temperature was maintained at 35℃; the detection wavelength was set at 203 nm. RESULTS: The calibration curves of ginsenoside Rg,, Re, Rb, and jujuboside A, B were in good linearity over 0.32-2.24 μg (r=0.9982), 0.16-1.12 μg (r=0.995 3), 0.32-2.24 μg (r=0.9996), 0.16-1.12 μg (r=0.9915), 0.08-0.56 μg (r=0.9996). The corresponding average recovery rates were 97.18%, 97.62%, 98.79%, 98.48%, 94.51%; the standard deviations were 1.10%, 0.98%, 0.34%, 1.09%, 1.88%, respectively. CONCLUSION: The established HPLC method is accurate, reliable and reproducible, which can be used as a reference for the quality control of RSJPW.
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To analyze and evaluate the quality of Ziziphi Spinosae Semen produced in Tianjin and comparing with Ziziphi Spinosae Semen traded on the market in different places. Twelve kinds of wild Ziziphi Spinosae Semen samples collected from the four represent regions were gathered by systematic combination of regional stratified sampling method and HPLC method was used to determine spinosin, jujuboside A, jujuboside B, and betulinic acid. The contents of spinosin, jujuboside A, jujuboside B, and betulinic acid from wild Ziziphi Spinosae Semen produced in Tianjin are 0.810-1.925, 0.695-1.708, 0.201-0.390, and 0.651-0.789 mg/g; Ziziphi Spinosae Semen produced in Chengde, Hebei province has the highest betulinic acid contents of 1.654 mg/g. The quality of wild Ziziphi Spinosae Semen produced in Tianjin is excellent. What's more, the wild Ziziphi Spinosae Semen produced in Baxian Mountain, Tianjin takes particularly prominent advantages of better quality. Quality of wild Ziziphi Spinosae Semen produced in other three regions is equal to the average level of that traded on the market.
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This article was aimed to study An-shen drug substances of semen ziziphi spinosae and the relationship between index component distribution in vivo and meridian tropism. Intragastric administration of thyroid tablet suspension at the dose of 160 mg·kg-1 was given for 13 days for the establishment of yin deficiency rat model. Elevated plus maze test (EPM) was combined with light/dark box test to evaluate the effect of semen ziziphi spinosae on anxiety behavior among yin deficiency rats. The yin deficiency anxiety model rats' eyeballs were picked for blood at 10, 20, 30, 40, 60, 90, 120, 240 min after intragastric administration of semen ziziphi spinosae decoction. The rats were sacrificed for the collecting of heart, liver, spleen, lungs, kidneys, stomach, brain, large intestine and small intestine and other tissues. HPLC-PDA-ELSD was combined to detect concentrations of spinosin, jujuboside A and jujuboside B in different tissues of rats. Related pharmacokinetic parameters were achieved after processing detected data with DAS 2.0 software. The results showed that compared with the yin deficiency group, semen ziziphi spinosae significantly reduced the rats' abnormal increased food-intake (P stomach > liver > brain > large intestine > spleen > lungs > heart > kidneys. The order of AUC0-t in tissues was small intestine > stomach > liver > large intestine > spleen > brain > heart > kidneys > lungs. The order of average concentration of jujuboside A among tissues was lungs > large intestine > heart > spleen > liver > kidneys > small intestine > stomach > brain. The order of AUC0-t in tissues was lungs > spleen > liver > heart > large intestine > brain > stomach > kidneys > small intestine. The order of average concentration of jujuboside B among tissues was large intestine > small intestine > stomach. The order of AUC0-t in tissues was large intestine > small intestine > stomach. It was concluded that semen ziziphi spinosae can obviously improve yin deficiency symptoms with good anti-anxiety effects. Spinosin and jujuboside A in semen ziziphi spinosae were the drug substances of An-shen effect. They were also the material basis of sweet and sour taste. The spinosin and jujuboside A distribution in vivo of yin deficiency anxiety model rats were close to the meridian tropism of semen ziziphi spinosae.
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This study was aimed to establish an analytical method to determine spinosin, jujuboside A, jujuboside B and betulinic acid content of Ziziphi Spinosae Semen quickly and accurately. The UPLC determination of Ziziphi Spinosae Semen method was established on an HSS T3 Column (2.1 mm × 100 mm, 1.8 μm) eluted with the mobile phase consisted of water containing 0.03% phosphoric acid and acetonitrile in gradient mode with the flow rate of 0.5 mL?min-1 and the detection wavelength was set at 204 nm. The standard curves of spinosin, jujuboside A, jujuboside B, betulinic acid showed a good linearity in 29.70 - 594.0 μg?mL-1, 10.68 - 213.6 μg?mL-1, 6.175 - 123.5 μg?mL-1, 22.00 - 440.0 μg?mL-1, respectively. The mean recoveries (n = 9) of the four components were between 98 . 3% and 99 . 4%, RSD < 1 . 4%. This method which is quick and accurate can be used to determine spinosin, jujuboside A, jujuboside B and betulinic acid in Ziziphi Spinosae Semen.