Discrimination of three medicinal materials from the Citrus genus by HPLC fingerprint coupled with two complementary software.

INTRODUCTION: Pericarpium Citri Reticulatae, Pericarpium Citri Reticulatae Viride and Fructus Aurantii come from the fruit of Citrus genus and possess analogous pharmacological activities. Either two or three of them are often present in one preparation. However, there is no standard method to differentiate the three herbal medicines for quality control. OBJECTIVE: To develop a fingerprint method for authentication of these three herbal medicines by HPLC. METHODOLOGY: Methanol extracts were analysed by HPLC, with a mobile phase of 0.1% phosphoric acid in water (A) and acetonitrile (B) in a gradient programme. The flow rate was set at 0.8 mL/min and UV detection at 320 nm. Principal component analysis and similarity evaluation were employed to analyse the chromatographic dataset. RESULTS: The chromatograms of 20 Pericarpium Citri Reticulatae, 13 Pericarpium Citri Reticulatae Viride and 15 Fructus Aurantii samples from diverse habitats had 13 peaks in common, and showed one peak characteristic for Pericarpium Citri Reticulatae Viride and two peaks characteristic for Fructus Aurantii. Furthermore, it was possible to differentiate the three medicines by a 'fingerprint region'. The difference between Pericarpium Citri Reticulatae and Fructus Aurantii was much bigger than that between Pericarpium Citri Reticulatae and Pericarpium Citri Reticulatae Viride, as could be shown by calculation of common peak ratio and variation peak ratio. CONCLUSION: A reliable HPLC fingerprint method coupled with principal component analysis and similarity evaluation was developed and showed substantial differentiation power for the three medicines.

A monoclonal-antibody-based ELISA for the detection of human FADD (Fas-associated death domain).

FADD (Fas-associated death domain) has been widely expressed in various tissues and its expression has been recently demonstrated to correlate with tumour progression and prognosis. Currently, measurement of FADD expression mainly depends on Western-blot or immunohistochemical approaches. To develop a conventional sandwich ELISA avenue for the detection of FADD protein to supplement Western blotting or immunohistochemistry, a series of mAbs (monoclonal antibodies) specific for FADD protein, designated 3A3, 3F9, 3G4, 4B9, 4G1, 7A8, 7B8 and 7F4, were produced by fusing mouse s/p20 myeloma cells with the spleen cells of a mouse immunized with the Escherichia coli-expressed recombinant His(6)-FADD protein. On the basis of the characterization of these mAbs, purified 3F9 was selected as the capture antibody and the biotin-conjugated 3A3 was selected as the detection antibody in sandwich ELISA. The limit of detection for the ELISA was 0.3 ng of purified His(6)-FADD (FADD tagged with hexahistidine), and it could detect both recombinant and native human FADD protein. Furthermore, the positive reaction of the ELISA could be blocked by rabbit anti-FADD sera. All of these results indicated that the ELISA developed in the present paper could be a promising tool for detection of FADD protein.

[Experimental study on anti-pyretic effect of gegen qin lian decoction and its compounds].

OBJECTIVE: To investigate composition principles of Gegen Qin Lian decoction through anti-pyretic experiment. METHOD: Pharmacological effects of different compounds of Gegen Qin Lian decoction according to six hours temperature response index (TRI6) and average top temperature response height (deltaT) after the decoction was given to feverish animal model by inactived bacteria suspension. RESULT: As for reducing six hour temperature response index, Scutellaria baicalensis root was the main effective drug. Pueraria lobata root could enforce the effect while Coptis chinensis rhizome and Glycyrrhiza uralensis root counteracted it. As for reducing average top temperature response height, the Effects of four herbal drugs were the same as for TRI6. CONCLUSION: Of the compounds of Gegen Qin Lian decoction, as to the pharmcological anti-pyretic effects, the best one is the compound of Scutellaria baicalensis and Pueraria lobata roots.