HPTLC Analysis of multi-ingredient traditional Chinese medicine supplement

OBJECTIVE Analysis of traditional Chinese medicinal(TCM)supplements has always been a laborious task,particularly in the case of multi-ingredient formulations.Traditionally,herbal extracts are analyzed using one or few markers compounds.In the recent years,however,pharmaceutical companies are introducing health supplements of TCM ingredients to cater to the needs of consumers in the fast-paced society in this age.As such,new problems arise in the aspects of composition identification as well as quality analysis.In most cases of products or supplements formulated with multiple TCM herbs,the chemical composition and nature of each raw material differs greatly from the others in the formulation.This results in a requirement for individual analytical processes in order to identify the marker compounds in the various botanicals.The aim of this study is to establish a rapid and low cost analytical approach using high performance thin layer chromatography(HPTLC)for the analysis of 3 key components of the LAC Liver Protector,a supplement with TCM formulation aimed at improving liver health.METHODS The TCM herbs were extracted with methods derived from Chinese Pharmacopeia standards.The herbal extracts were then applied to HPTLC plates using CAMAG Linomat 5 and developed using fully saturated twin-trough chromatographic chambers.The developed plates were derivatized and documented with CAMAG TLC Visualizer.The images were processed with CAMAG winCATS software to calculate the Rf values and confirm the presence of key constituents of the herbs.RESULTS The 3 key components of LAC Liver Protector product,namely Radix notoginseng,Rhizoma alismatis and Radix paeoniae alba/rubra were identified in all 6 batches of products tested.CONCLUSION With the increasing trend of small and medium-sized enterprises(SMEs)bringing natural products and health supplements into the market,it is crucial that the qualities of both raw materials and end products be well-assured for the protection of consumers.With the technology of HPTLC,science can be incorporated to help SMEs with their quality control,thereby ensuring product quality.

In vitro and in vivoimmunomodulatory activities of polysaccharides purified from four species of Dendrobium

OBJECTIVE To investigate the isolation,extraction,purification,and immunomodulatory activities in vitro and in vivo of polysaccharides from four different species of Dendrobiumherbs widely used in Singapore.METHODS To apply the classic method of water extraction and ethanol precipitation and the gel column chromatography technique,the polysaccharides from four species of Dendrobium herbs(D.moniliforme,D.crystallinum,D.primulinum,D.chrysanthum)were obtained.The polysaccharides were tested first on RAW 264.7 cell line with MTT method to test the effect of polysaccharides on macrophages.The two polysaccharides from D.moniliforme were found to promote proliferation of the cells,and were then tested on mice splenocyte primary cell culture.The same polysaccharides were also injected to female BALB/c mice at various doses for 21 dto see whether they would have affected the organs,especially the livers,spleens and thymuses.RESULTS Out of the 4 Dendrobiumspecies tested,it was found that polysaccharides from D.moniliformeto be most supportive of the proliferation of macrophages in a dose-dependent manner,with increasing effect with the increasing doses used.Polysaccharides from D.chrysanthumand D.crystallinum were also found to act similarly albeit to a lesser extent.The two D.moniliforme polysaccharides were also found to greatly enhance the proliferation of mouse splenocytes in vitro and growth of the spleen in vivo.CONCLUSION The immunomodulatory activities in vitro and in vivo of polysaccharides from four species of Dendrobium herbs were different.Their diverse chemical characters might contribute to their varied bioactivity,which should be further investigated.In addition,it was suggested that the polysaccharides from D.moniliforme could be potential immunostimulants used in the healthcare and food industry.

Human urine metabolome in response to Fermentum rubrum(Hongqu Capsules)by HILIC-UHPLC-QTOF

OBJECTIVE ″-omics″study represents an unbiased perspective to examine the bio-system to discover the novel biomarker(s)which might be overlooked when targeted analysis is performed instead.Urine,due to its ease of collection,minimal invasion involved and rich information of the downstream metabolites,has been extensively studied.Fermentum rubrum(Hongqu,HQ capsules)is a well-known traditional medicine with the claimed slimming effect which could be related to the reduction in the deposition of total cholesterol and glycerides.Lack in sufficient clinical evidence is always one of the leading reasons that hamper traditional medicines from gaining world-wide recognition.In our pursuit of scientific support for HQ capsules in managing obesity,we aim to examine the alteration of urinary metabolites in this small-scale human clinical research.METHODS 6 Chinese subjects were included and exposed to short-term administration of HQ capsules for 3d,twice daily,two capsules each dosing.The urine samples were collected for three days prior to the dosing and on the day following the last dosing.In total,96 urine samples were collected and then separated on hydrophilic interaction chromatography(HILIC)and analyzed by Agilent 6550 ESI-Quadrupoletime-of-flight(QTOF)mass spectrometer.RESULTS Under positive mode,two compounds were found to be present only in post-dosing urine,and one compound was significantly lowered in post-dosing samples.Those two compounds might be associated with the administered HQ capsules,which is formulated with multiherbal constituents.CONCLUSION Further elucidation on the structures of these compounds is needed to enable better understanding of the mechanism of HQ capsules in managing obesity.

Gut microbiome in acute pancreatitis:A review based on current literature

The gut microbiome is a complex microbial community,recognized for its potential role in physiology,health,and disease.The available evidence supports the role of gut dysbiosis in pancreatic disorders,including acute pancreatitis(AP).In AP,the presence of gut barrier damage resulting in increased mucosal permeability may lead to translocation of intestinal bacteria,necrosis of pancreatic and peripancreatic tissue,and infection,often accompanied by multiple organ dysfunction syndrome.Preserving gut microbial homeostasis may reduce the systemic effects of AP.A growing body of evidence suggests the possible involvement of the gut microbiome in various pancreatic diseases,including AP.This review discusses the possible role of the gut microbiome in AP.It highlights AP treatment and supplementation with prebiotics,synbiotics,and probiotics to maintain gastrointestinal microbial balance and effectively reduce hospitalization,morbidity and mortality in an early phase.It also addresses novel therapeutic areas in the gut microbiome,personalized treatment,and provides a roadmap of human microbial contributions to AP that have potential clinical benefit.

Qualitative and quantitative analysis of Arnebiae Radix and Dictamni Cortex and efficacy study of herbal extracts on allergic contact dermatitis using 3D human reconstructed epidermis

Objective:To evaluate the quality of Arnebiae Radix(AR)and Dictamni Cortex(DC)and study the efficacy of herbal extracts of these two herbs on the treatment of allergic contact dermatitis(ACD).Methods:Qualitative and quantitative analysis of effective components was performed using High Performance Thin Layer Chromatography(HPTLC),High Performance Liquid Chromatography(HPLC),and HPLC-Quadrupole Time of Flight-Mass Spectrometry(HPLC-QTOF-MS).In vitro allergic ACD 3D model was established by incubating 3D reconstructed human epidermis(RHE)with skin sensitizer,potassium dichromate.A total of 65 gene expression that were associated with ACD,which included 24 antioxidant responsive element(ARE)and 41 SENS-IS genes were quantified by q RT-PCR.More than or equal to 10 ARE genes and 18 SENN-IS genes were induced by 1.3-fold,demonstrating the successful establishment of in vitro ACD model.Oil extracts of AR and DC were applied on the in vitro ACD model to study the efficacy.Results:Batch 3 of AR and batch 2 of DC showed presence of all active ingredients with the highest concentrations.Active ingredients of the herbs were extracted using a special oil and formulated into herbal oil extracts.The herbal oil extracts were able to down regulate the induced genes in the in-vitro ACD skin model,bringing the tissue back to homeostatic status.Conclusion:The oil extracts showed the potent efficacy of using AR and DC in ACD treatment.The combination study will be done to optimize the formulation ratio which will be developed into a topical cream.