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Objective:To establish reverse-phase high-performance liquid chromatography and simultaneously determine gallic acid, methyl gallate, corilagin, Sennoside B, and Sennoside A levels in Sana preparations.Methods:From January to December 2021, Phenomenex Hydro-RP 80A column (4.60 mm × 250 mm, 4 μm) was used. Elution was conducted using mobile phases methanol (A)-0.2% formic acid (B). The following gradients were applied: 1%-3%A for 0-18 minutes, 3%-15%A for 18-19 minutes, 15%-17%A for 19-40 minutes, 17%-25%A for 40-45 minutes, 25%-35%A for 45-65 minutes, 35%-60%A for 65-95 minutes, 60%-90%A for 95-96 minutes, 90%-1%A for 96-97 minutes. The flow rate was 1.0 mL/minute. The column temperature was 35 ℃. The detection wavelength was 270 nm.Results:The linear ranges of gallic acid, methyl gallate, corilagin, Sennoside B and Sennoside A were 0.182-1.099 μg ( r = 0.999 9), 0.046-0.278 μg ( r = 0.999 2), 0.266-1.598 μg ( r = 0.999 4), 0.172-1.036 μg ( r = 0.999 2), and 0.176-1.056 μg ( r = 0.999 9). The average dosing recovery rates were 100.02%, 99.14%, 99.38%, 101.77%, and 100.92%, respectively. Conclusion:Reverse-phase high-performance liquid chromatography can be used for quality control of Sana preparations because of high accuracy, sensitivity, reliability, and reproduction.
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Objective: Sennosides A and B, which are dianthrone glycosides contained in Rhubarb and Senna Leaf, exhibit laxative effect. Although a number of over-the-counter (OTC) drugs used as laxatives contain Rhubarb or Rhubarb and Senna Leaf, the total amounts of sennosides A and B are not mentioned in the package insert. To determine the total amounts of sennosides A and B in OTC drugs containing Rhubarb or Rhubarb and Senna Leaf, quantitative analyses of sennosides A and B were performed for 24 OTC drugs.Methods: Sennosides A and B were extracted from 24 OTC drugs and quantitatively analyzed by high-performance liquid chromatography. Statistical analyses were carried out by a one-way analysis of variance followed by Dunnett's test or Tukey's test.Results: The OTC drugs contained sennosides A and B in the range of 1.5-10 mg in the minimum daily dosage and in the range of 2.7-17 mg in the maximum daily dosage. In 11 of the OTC drugs (Products Nos. 1-5, 11, 12, and 15-18), the maximum daily dosage contained almost equal or higher amounts of sennosides A and B compared to that in a tablet of the prescription medicine Pursennid® 12 mg. Furthermore, the amounts of sennosides A and B in the maximum daily dosage were significantly higher in products Nos. 1 and 11 and lower in products Nos. 8-10, 14, and 20-24 compared to those of a tablet of Pursennid® 12 mg.Conclusion: Although some OTC drugs have the same Rhubarb content, the total amounts of sennosides A and B can vary. Thus,there is no correlation between the Rhubarb content and total amounts of sennosides A and B. This is because of the inconsistent quality of Rhubarb and/or the differences in the manufacturing methods of the OTC drugs containing Rhubarb. Because the total amounts of sennosides A and B cannot be estimated based on the Rhubarb content, a constipated patient should start taking an OTC drug containing Rhubarb at the minimum daily dosage. It is also recommended that the total amounts of sennosides A and B are mentioned in the package insert of OTC drugs containing Rhubarb or Rhubarb and Senna Leaf.
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OBJECTIVE Senna and rhubarb are classified as stimulative laxatives, and known to have similar effec?tive constituents, the anthraquinones. Being protected by theβ-glucoside bond, the anthraquinones can reach the intes?tines where they are degraded into complex metabolites by enzymes secreted from the intestinal microbiome. It is these complex metabolites that produce the laxative effects. Then the similarities and differences of action between the anthra?quinones require further elucidation. METHODS Here, we studied metabolites of senna anthraquinones (SAQ), rhubarb anthraquinones (RAQ) and their chemical marker, sennoside A (SA), in a rat diarrhea model. In the in vitro biotransfor?mation experiments, SAQ, RAQ and SA were incubated with rat fecal flora solution and the metabolites produced were analyzed using HPLC. In the in vivo studies, the same compounds were investigated for purgation induction, with mea?surement of histopathology and multiple aquaporins (Aqps) gene expression in six organs. RESULTS SAQ and RAQ had similar principal constituents but could be degraded into different metabolites. A similar profile of Aqps down-regula?tion for all compounds was seen in the colon, suggesting a similar mechanism of action for purgation. However, in the kidneys and livers of the diarrhea-rats, down-regulation of Aqps was found in the RAQ-rats whereas up-regulation of Aqps was seen in the SAQ-rats. Furthermore, the RAQ-rats showed lower aquaporin 2 (Aqp2) protein expression in the kidneys, whilst the SA-rats and SAQ-rats had higher Aqp2 protein expression in the kidneys. This may have implications for side effects of SAQ or RAQ in patients with chronic kidney or liver diseases. CONCLUSION SAQ and RAQ showed similar laxative actions with a similar mechanism, they could display different actions in rat kidneys and livers. We suggest that the clinical usage of senna or rhubarb products should be clarified for patients having chronic kidney or liver diseases.
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OBJECTIVE： To establish a method for simultaneous determination of 8 non-anthraquinone constituents in Rheum palmatum. METHODS： HPLC method was adopted. The determination was performed on Symmetry C18 column with mobile phase consisted of methanol-0.1％ phosphoric acid （gradient elution） at the flow rate of 1.0 mL/min. The column temperature was 30 ℃ and detection wavelength was 280 nm. Sample size was 30 μL. RESULTS： The linear range of gallic acid， catechin， epicatechin， resveratrol 4′-O-glucopyranoside， epicatechin gallate， resveratrol 4′-O-β-D-（6″-O-galloyl）-glucopyranoside， sennoside A， 4′-hydroxyphenyl-2-butanone-4′-O-β-D-（2″-O-galloyl-6″-O-p-hydroxy cinnamyl）-glucopyranoside were 6.16-2 464 ng（r＝0.999 9）， 37.4-14 960 ng（r＝0.999 9）， 7.635-3 054 ng（r＝0.999 7）， 7.63-3 052 ng（r＝0.999 9）， 8.32-3 328 ng（r＝0.999 9）， 11.5-4 600 ng（r＝0.999 9）， 16.08-6 432 ng（r＝0.999 9）， 29.3-11 720 ng（r＝0.999 9）， respectively. The limits of quantitation were 3.48， 4.30， 6.40， 4.40， 3.39， 2.87， 8.40 and 4.95 ng， respectively. The limits of detection were 2.32， 2.58， 2.40， 2.64， 2.26， 1.23， 4.20， 2.97 ng， respectively. RSDs of precision， stability and reproducibility tests were all lower than 5％. Recoveries were 94.32％- 100.54％（RSD＝2.78％，n＝6）， 91.15％-99.36％（RSD＝3.72％，n＝6）， 92.16％-98.04％（RSD＝2.39％，n＝6）， 93.41％-100.73％（RSD＝3.17％，n＝6）， 93.89％-98.40％（RSD＝1.99％，n＝6）， 92.61％-101.74％（RSD＝3.71％，n＝6）， 92.66％-103.40％（RSD＝3.76％，n＝6）， 95.45％-102.70％（RSD＝3.06％，n＝6）， respectively. CONCLUSIONS： The established method is simple， accurate and specific， and can be used for the simultaneous determination of 8 non-anthraquinone constituents in R. palmatum.
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Sennoside A (SA) is a bioactive component of Chinese herbal medicines with an activity of irritant laxative, which is often used in the treatment of constipation and obesity. However, its activity remains unknown in the regulation of insulin sensitivity. In this study, the impact of SA on insulin sensitivity was tested in high fat diet (HFD)-induced obese mice through dietary supplementation. At a dosage of 30 mg/kg/day, SA improved insulin sensitivity in the mice after 8-week treatment as indicated by HOMA-IR (homeostatic model assessment for insulin resistance) and glucose tolerance test (GTT). SA restored plasma level of glucagon-like peptide 1 (GLP1) by 90% and mRNA expression of by 80% in the large intestine of HFD mice. In the mechanism, SA restored the gut microbiota profile, short chain fatty acids (SCFAs), and mucosal structure in the colon. A mitochondrial stress was observed in the enterocytes of HFD mice with ATP elevation, structural damage, and complex dysfunction. The mitochondrial response was induced in enterocytes by the dietary fat as the same responses were induced by palmitic acid in the cell culture. The mitochondrial response was inhibited in HFD mice by SA treatment. These data suggest that SA may restore the function of microbiota-GLP1 axis to improve glucose metabolism in the obese mice.
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Objective To develop an HPLC method to determine the contents of chrysophanol, emodin, physcion, aloe-emodin, sennoside A, and sennoside B simultaneously in the aerial part of Rheum tanguticum Maxim.ex Balf. Methods The analysis was achieved with an Agilent ZORBAX SB-C18 analytic column (250 mm × 4.6 mm, 5 μm) by gradient elution of methanol-0.1％ phosphoric acid in water gradient elution system (0 min, 70％ B; 5 min, 65％ B;8–16 min, 60％ B; 18–23 min, 55％ B; 25–30 min, 45％ B; 32–39 min, 35％ B; 49–57 min, 24％ B; 65–72 min, 20％ B). The flow rate was 1.0 mL/min; detection wavelength was 254 nm; the column temperature was room temperature; the quantification used external standard method. Results The peak areas and concentrations of chrysophanol, emodin, physcion, aloe-emodin, sennoside A, and sennoside B showed good linear relationship within a certain concentration range (r≥0.9995); the RSD of precision was 0.75％–1.17％; the RSD of repeatability was 0.99％–2.06％; the RSD of stability was 0.97％–1.76％; the average recoveries were 99.7％–100.4％. The results showed that there were differences in content between the root and aerial part of Rheum tanguticum Maxim.ex Balf. Conclusion HPLC method for simultaneous determination of contents of 6 contents of the aerial part of Rheum tanguticum Maxim.ex Balf can be used as the references for quality control.
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Sennoside A (erythro) and sennoside B (threo) are dianthrone glycosides and diastereomers. We investigated their abilities to prevent the gastric lesions associated with diseases, such as, gastritis and gastric ulcer. To elucidate their gastroprotective effects, the inhibitions of HCl*EtOH-induced gastritis and indomethacin-induced gastric ulcers were assessed in rats. It was observed that both sennoside A and sennoside B increased prostaglandin E2 (PGE2) levels and inhibited H+/K+-ATPase (proton pump). In a rat model, both compounds reduced gastric juice, total acidity and increased pH, indicating that proton pump inhibition reduces gastric acid secretion. Furthermore, sennoside A and B increased PGE2 in a concentration-dependent manner. In a gastric emptying and intestinal transporting rate experiment, both sennoside A and sennoside B accelerated motility. Our results thus suggest that sennoside A and sennoside B possess significant gastroprotective activities and they might be useful for the treatment of gastric disease.