| ObjectiveTo establish a comprehensive, scientific, reliable, and adaptable methods for evaluation and quality control of Guifuzhitong cataplasm, and followed by evaluation of its therapeutic effects on osteoarthritis, rheumatism and kidney tonic.MethodsThin layer chromatography was applied to identify radix dipsaci, clematis root, mulberry parasitism, radix zanthoxyli. In the formula, content determinations was carried out use HPLC, and were quantitatively analyzed for quality control of active compounds in the ingredients. At the same time, limits of double-ester alkaloids were also determined to ensure such toxic compounds were in acceptable ranges. Other items were tested according to regulations on cataplasm in "China pharmacopeia" (edition 2010). For the part of pharmacology, acetic acid caused mice pain model, radiant heat mice model and hot plate and electrical stimulation mice model were used. Acute toxicity and skin irritation was carried out in rabbit model and guinea pig was used to evaluate skin hypersensitivity.ResultsAs developed TLC chromatograms indicated, dots of radix dipsaci, clematis root, mulberry parasitism and radix zanthoxyli in preparation was clear without interferences in negative references, proving that the optimized TLC method can be used as identification. For content determination, the HPLC method used successfully determined concentrations of Guifuzhitong cataplasm, which were cinnamic aldehyde is 0.33 mg/stick, benzoylmesaconine 2.25 mg/stick, benzoylaconitine 0.83 mg/stick and benzoylhypacoitine 2.51 mg/stick, respectively; and double-ester alkaloids were not detected in the preparation. Results of general principle of preparation showed that paste content was 9.58g/cm2, ball 13 was stocked by the cataplasm in adhesion measurement; weigh difference were within ±5%. Results of microorganism showed that the limits of bacteria, mould and yeast were less than 100cfu/10cm2, and staphylococcus aureus and pseudomonas aeruginosa were not detected. Significant deterioration was observed in 18 months, indicating that the stability of the preparation was good.For pharmacology experiments, nociception tests mentioned above showed that the preparation has good analgesia effects in both medium and high dosage (p<0.05), which proved that Guifuzhitong cataplasm has good effect in analgesia.Results in acute toxicity, skin irritation and skin hypersensitivity tests showed no obvious effects, therefore, the safety of this preparation was guaranteed.Conclusions:This research proposed a comprehensive, scientific and reliable method for evaluation of Guifuzhitong cataplasm and results of identification and quantitative determination proved the stability and controllability of the preparation. In addition, the pharmacological experiment was effective and suitable for evaluation of its therapeutics on osteoarthritis. In general, the above mentioned researches built the foundation and reference for experimental production and pre-clinical research of Guifuzhitong cataplasm. |
