Quantitative Analyses of Sennosides A and B in Over-the-Counter Drugs Containing Rhubarb or Rhubarb and Senna Leaf / 医薬品情報学

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.

Simultaneous determination of five active ingredients including gallic acid in different dosage forms of Sana preparation by reverse-phase high-performance liquid chromatography / 中国基层医药

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.

Organ specific differences in alteration of aquaporin expression in rats treated with sennoside A, senna anthraquinones and rhubarb anthraquinones / 中国药理学与毒理学杂志

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.

Simultaneous Determination of 8 Non-anthraquinone Constituents in Rheum palmatum by HPLC / 中国药房

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.

Regulation of microbiota-GLP1 axis by sennoside A in diet-induced obese mice

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.

Multiwavelength detection and multivariate analysis for tongzhi surunjiang preparation with substitute reference substance method by high performance liquid chromatography / 中国药学杂志

OBJECTIVE: To propose a holistic strategy for quality control of Chinese patent medicines, and establish an HPLC analytical method for Tongzhi Surunjiang preparation according to the strategy. METHODS: The strategy contained three steps.The first step was multi-wavelength chromatographic detection.The second step was multivariate analysis for identification and assay. The third step was to establish substitute reference substance method.The preparations were extracted by ultrasound with methanol, chromatography was performed on ODS column with gradient elution with acetonitrile and 0.1%phosphoric acid aqueous solution.The detection wavelengths were set at 270, 350, 410 and 440 nm.The radar chart, HCA heatmap, principal component analysis and cosine similarity were used for data analysis.At last, linear calibration using two reference substances, relative retention time method and PDA spectrum method were used for peak identification, and relative correction factor method was used for quantitative analysis. RESULTS: The multi-components determination method and fingerprint analysis met the method validation requirements. Data analysis showed that there were some differences among the samples of different manufacturers. Strong characteristic peaks for classification were gallic acid, chebulinic acid, chebulea fructus, sennae folium, sennae folium and crocin.CONCLUSION: The method is specific, with low cost, and could be used to accurately control the quality of Tongzhi Surunjiang preparation.

Simultaneous Determination of Contents of 6 Components in Aerial Part of Rheum tanguticum Maxim.ex Balf by HPLC Method / 中国中医药信息杂志

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.

Simultaneous Determination of 10 Kinds of Chemical Components in Processed Products of Rhei Radix et Rhizoma / 中国药房

OBJECTIVE:To establish a method for the contents of gallic acid,catechin,sennosides B,aloe-emodin,rhein, emodin,chrysophanol,physcion,chrysophanol-1-O- glucoside and emodin-8-O- glucoside in Rhei Radix et Rhizoma,Jiu Rhei Radix et Rhizoma,Shu Rhei Radix et Rhizoma,Rhei Radix et Rhizoma tan,Cu Rhei Radix et Rhizoma,and analyze the differ-ences. METHODS:HPLC was performed on the column was Hypersil C18 with mobile phase of methanol- 0.2% acetic acid(gradi-ent elution)at a flow rate of 1.0 ml/min,the detection wavelength was 260 nm,column temperature was 25 ℃,injection volume was 10 μl. RESULTS:The linear range was 0.252 5-4.040 0 μg for gallic acid(r=0.999 6),0.600 0-9.600 0 μg for catechin(r=0.999 6),0.297 4-4.758 4 μg for sennosides B(r=0.999 9),0.001 8-0.028 8 μg for aloe-emodin(r=0.999 9),0.005 0-0.080 0 μg for rhein(r=0.999 9),0.019 0-0.304 0μg for emodin(r=0.999 8),0.380 2-6.083 2μg for chrysophanol(r=0.999 7),0.008 2-0.131 2μg for physcion(r=0.999 8),0.126 0-2.016 0 μg for chrysophanol-1-O-glucoside(r=0.999 6)and 0.111 3-1.780 8 μg for emo-din-8-O-glucoside (r=0.999 8);RSDs of precision,stability and reproducibility tests were lower than 3.0%;recoveries were 96.17%-97.21%(RSD=1.67%,n=6),97.60%-100.54%(RSD=2.55%,n=6),99.45%-101.32%(RSD=1.63%,n=6), 95.31%-98.19%(RSD=2.42%,n=6),98.99%-100.35%(RSD=1.86%,n=6),98.95%-101.21%(RSD=2.17%,n=6), 99.81%-100.62%(RSD=1.66%,n=6),96.78%-98.52%(RSD=1.99%,n=6),97.80%-100.14%(RSD=3.32%,n=6) and 97.40%-101.24%(RSD=2.89%,n=6). Compared with Sheng Rhei Radix et Rhizoma,the contents of gallic acid,catechin,sen-nosides B and anthraquinones in Cu Rhei Radix et Rhizoma,Jiu Rhei Radix et Rhizoma and Rhei Radix et Rhizoma tan decreased. The contents of catechin,sennosides B and anthraquinones in Shu Rhei Radix et Rhizoma. Catechin,sennosides B,chrysopha-nol-1-O- glucoside,aloe-emodin and rhein were not detected in Dahuang tan. CONCLUSIONS:The method is simple with good precision,stability and reroducibility,and can be used for the simultaneous determination of 10 chemical components in processed products of Rhei Radix et Rhizoma;there were significant differences in contents of 10 chemical components in processed prod-ucts of Rhei Radix et Rhizoma.

Gastroprotective Activities of Sennoside A and Sennoside B via the Up-Regulation of Prostaglandin E2 and the Inhibition of H+/K+-ATPase

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.

Simultaneous determination of eight components in Hedan Tablets by UPLC / 中草药

Objective: To develop a UPLC method for the simultaneous determination of nuciferine, psoralen, isopsoralen, tanshinone IIA, danshensu, sennoside A, sennoside B, and ursolic acid in Hedan Tablets. Methods: The chromatographic separation was achieved on an Acquity UPLC BEH C18 column (100 mm × 2.1 mm, 1.7 μm) with methanol-acetonitrile (40:60, A)-0.1% formic acid (B) as mobile phases at the flow rate of 0.6 mL/min for gradient elution: 0-4.0 min, 90% B; 4.0-6.0 min, 90%-80% B; 6.0-9.0 min, 80%-70% B; 9.0-10.5 min, 70%-60% B; 10.5-12.0 min, 60%-50% B, 12.0-16.0 min, 50%-10% B; Detection with variable wavelength: 0-4.0 min was 215 nm, 4.0-12.0 min was 270 nm, 12.0-16.0 min was 340 nm, and the column temperature was 40°C. Its linear relationship, precision, repeatability, stability, and recoveries were investigated. Results: The results showed that the eight active components were well separated and showed good linearity. The precision was good and RSD was less than 3.0%. The repeatability was good and RSD was less than 3.0%. The stability was good in 24 h. The average recoveries were between 99.21%-101.91% and RSD was less than 3.0%. Conclusion: The method is accurate, sensitive, credible, and repeatable. It could be applied to the quality control of Hedan Tablets.

Determination of anthraquinones and bianthrones in rhubarb from different sources by HPLC / 中草药

Objective: To develop a HPLC method for determining the content of aloe-emodin, rhein, emodin, chrysophanol, physcion, sennoside A, and sennoside B simultaneously. The method was used in content determination of constituents in rhubarb from different sources. Methods: The separations were carried out at 30°C on an Agilent Eclipse XDB-C18 column (150 mm x 4.6 mm, 5μm) and eluted with acetonitril and water containing 0.1% phosphoric acid in gradient mode. The flow rate was 1.0 mL/min, detection wavelengths were 254 nm for aloe-emodin, rhein, emodin, chrysophanol, physcion and 340 nm for sennoside A and sennoside B, column temperature was 30°C. Results: The peak areas and injection ammounts of aloe-emodin, rhein, emodin, chrysophanol, physcion, sennoside A and sennoside B showed a good linear relationship within a certain range. The average recoveries were standards-compliant. The method is simple, accurate and repeatable to be used in content determination of rhubarb. Conclusion: The total anthraquinones content and total sennosides content in cultivated samples are much less than those in the wild samples.

A Herbological Study on the Medicinal Effects and Employments of Rhubarb Processed with Liquor (2) / 日本東洋医学雑誌

In traditional Chinese medicine, crude drugs are sometime processed and prepared for specific purposes. Rhubarb (Da-huang in Chinese; Daio in Japanese) has been processed by dipping or soaking it in huangjiu (Chinese fermented wine). However, the pharmacologic significance of this liquor processing has not been elucidated thoroughly. In this report, we describe how processing with ethanol altered the levels of the principal compounds in rhubarb: sennoside A, sennoside B, aloe-emodin, rhein, emodin, chrysophanol, physcion, lindleyin, isolindleyin, and total tannins. Liquor-dipping, dipping rhubarb in 16% ethanol for 30 seconds, did not affect the content of sennosides. Thus, the purgative effect of rhubarb is likely to be preserved. Liquor-soaking, soaking rhubarb in ethanol for 12 to 24 hours, the content of sennosides and tannins decreased and the content of anthraquinones increased. Liquor-soaking of rhubarb may increase its anti-bacterial and anti-inflammatory effects, thereby improving blood stasis. These results are in agreement with the descriptions in medicinal literatures published since the Jin and Yuan Dynasties.

Clinical Findings in Chronic Renal Failure by Administering Wen-Pi-Tang (Onpito) Specially Compounded with Processed Rhubarb / 日本東洋医学雑誌

Rhubarb has been reported effective in patients with chronic renal failure, especially those patients with a high blood urea nitrogen (BUN) value. Rhubarb, however, is difficult to administer in large quantities because of its purging effect. Thus we prepared heated rhubarb, measured its sennoside A content using high performance liquid chromatography, and administered it to 2 patients.<br>Renal function of the first patient did not improve, following administration of extract derived from decocted rhubarb heated for 4 hours at 100°C.The extract's protective effect on renal function was considered lost by the boiling procedure. Dry-heated rhubarb was also prepared at 130°C for 4 minutes, and administered to the second patient. In this case, the extract's quantity could be increased up to 4.0g per day, and the patient's BUN value gradually decreased.<br>Onpito was later administered, because of increased BUN and creatinine values. Onpito was administered as a Ninjinto extract, Kakobushimatsu and rhubarb. And Onpito had a better protective effect on renal function in the second patient, than rhubarb alone.<br>In conclusion, we believe that dry-heated rhubarb can be effective for the improvement of nitric metabolism in patients with chronic renal failure, because of its low purging effect.