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.

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.

A fast screening method for multi-residue pesticide analysis of TCM herbs using liquid chromatography-quadrupole-time-of-flightmass spectrometry

OBJECTIVE Identification of pesticide residues in TCM herbs is important for drug safety and public health.However,it is very challenging for the development of an instrument-based multi-residue pesticide analysis method for theherbalproducts due to their complex matrix.This study aims to develop a rapid method for the high-throughput detection of pesticide residues in TCM herbal products using liquid chromatography coupled to quadrupoletime of flight(LC-QTOF)mass spectrometry.METHODS In this study,we attempt to investigate possible pesticide residues in TCM herbal products which are commonly available in Singapore and to consolidate a comprehensive pesticide database for more than 250 regulated pesticides in FDA Regulations.We also attempt to perform non-targeted and targeted screenings of pesticide residues in herbal products by searching our database.RESULTS The developed LC-QTOF method demonstrates have high mass accuracy,resolution and selectivity,which makes it a very suitable tool for monitoring both target and non-target pesticides with a limit of detection at the low ppb level.Moreover,the LC-QTOF technology permits to retain all spectral data in a single run.It may avoid many difficulties related to matrix effects and will save much manpower.CONCLUSIONThis means that a variety of TCM herbs can be easily detected by this method for the presence or absence of pesticide residues that must conform to government regulations.

Expression of human TNF-related apoptosis-inducing ligand extracellular region in E. coli

This study is conducted to clone the cDNA encoding human TNF-related apoptosis-inducing ligand (hTRAIL) extracellular region (amino acids 41-281, hTRAIL41-281) and to express it in E.coli. The hTRAIL41-281 cDNA is amplified by reverse transcription (RT) PCR from total RNA derived from human acute promyelocytic leukemia cell line HL-60. After sequenced, the cDNA is cloned into the vector pQE-80L and transformed into E.coli DH5 to express the recombinant hTRAIL41-281 (rhTRAIL41-281) induced by IPTG. The recombinant protein is analyzed by SDS-PAGE. The cloned cDNA is consistent with the cDNA sequence encoding hTRAIL41-281 reported in GenBankTM. After inducing, the hTRAIL41-281 protein is expressed, and the mass of the recombinant protein is about 30 % of total bacteria protein, which demonstrates that the cDNA encoding hTRAIL41-281 is successfully cloned and expressed in E.coli.

Automorphisms of Maps with a Given Underlying Graph and Their Application to Enumeration

A graph is called a semi-regular graph if its automorphism group action onits ordered pair of adjacent vertices is semi-regular. In this paper, a necessary and sufficientcondition for an automorphism of the graph Γ to be an automorphism of a map with the underlyinggraph Γ is obtained. Using this result, all orientation-preserving automorphisms of maps onsurfaces (orientable and non-orientable) or just orientable surfaces with a given underlyingsemi-regular graph Γ are determined. Formulas for the numbers of non-equivalent embeddings of thiskind of graphs on surfaces (orientable, non-orientable or both) are established, and especially, thenon-equivalent embeddings of circulant graphs of a prime order on orientable, non-orientable andgeneral surfaces are enumerated.

On κ-ordered Graphs Involved Degree Sum

Abstract A graph g is k-ordered Hamiltonian, 2 h k h n, if for every ordered sequence S of k distinct vertices of G, there exists a Hamiltonian cycle that encounters S in the given order. In this article, we prove that if G is a graph on n vertices with degree sum of nonadjacent vertices at least $n+{{3k - 9} \over 2}$, then G is k-ordered Hamiltonian for k=3,4,...,￠${n \over {19}}$?. We also show that the degree sum bound can be reduced to n+ 2 ￠ ${k \over {2}}$ m 2 if $\kappa(G)\ge {{3k - 1} \over 2}$ or '(G) S 5k m 4. Several known results are generalized.