Preparation, Quality Analysis And Activity Evaluation Of An Traditional Chinese Medicated Oil With Anti-inflammatory And Analgesic Effect

Traditional Chinese Medicated Oil has characteristics of percutaneous absorption, fast onset, and less side effects (gastrointestinal irritation etc.). However, there is few reports on the pharmacological activity of Traditional Chinese Medicated Oil, and no clear process quality standards was found currently. By analyzing the ingredient of medicated oil products in market and pharmacological studies on the herbs in these formulae, volatile oils (referred to as V) were extracted from Angelicae Pubescentis Radix, myrrh, Curcumae Rhizoma and Schizonepetae Herba by the extraction-rectification coupled device. Similarly, saponins (referred to as S) were isolated from Pubescent Holly Root, Himalayan Teasel Root and Chinese clematis root by solvent reflux extraction coupled macroporous resin adsorption, when the safflower flavonoids (referred to as F) was obtained by ultrasonic water extraction combined with macroporous resin purified. Nitidine alkaloids (eferred to as A) were extracted by heating reflux with enthol and solvent extraction. The UV spectrophotometry (UV) methods for detecting the content of total saponins of Pubescent Holly Root, Himalayan Teasel Root and Chinese clematis root were established, and the high performance liquid chromatography (HPLC) methods for detecting the content of Hydroxyl Safflor Yellow A (HSYA) in safflower flavonoids and nitidine chloride in nitidum alkaloids were set up at the same time. The gas chromatography-mass spectrometry (GC-MS) methods for analysis the volatile oil of Angelicae Pubescentis Radix, myrrh, Curcumae Rhizoma and Schizonepetae Herba were developed as well. Then herbal extracts V, S, F, A were formulated into the sample of medicated oil (HLYA) with wintergreen oil, camphor and menthol. In addition, we preparation four disassembled formulae samples (V, VF, VA, VFA, VAFS), and the disassemble samples contained the herbal extracts V, V+F, V+A, V+A+F and V+A+F+S, respectively. For each sample, the anti-nociceptive activity was evaluated by the hot plate test and acetic acid writhing in mice, when the anti-inflammatory activity was evaluated by ear edema model in mice. Thin layer chromatography (TLC) was applied for qualitative analysis on HLYA. Afterwords. HPLC and GC-MS were used for quantitative analysis and quality analysis of chlorinated nitidine and volatile oils contained in HLYA, respectively.The results showed that, the volatile oil of curcuma mainly contains eucalyptol (17.78%). curcuma ketone (37.86%). p-elemene (8.54%). etc.. Schizonepetae volatile oil mainly contains menthone (48.42%) and pulegone (43.01%), etc.. Angelicae pubescentis volatile oil mainly contains:a-Thuja ene (22.86%), p-Thuja ene (24.96%), etc.. Myrrh volatile oil mainly contains8-elemene (15.58%), p-elemene (12.98%), octyl acetate (39.33%), and so on. Total saponins content of Pubescent Holly Root, Himalayan Teasel Root and Chinese clematis root were51.21%,16.31%and25.69%, respectively. The content of HSYA in safflower flavonoids was21.60%, and the nitidine chloride content in nitidum alkaloids was1.56%. The TLC results show that HLYA contains extracts all from Pubescent Holly Root, Himalayan Teasel Root, Chinese clematis root, safflower, Schizonepetae Herba and Curcumae Rhizoma. The quantitative analysis result showed that HLYA containing nitidine chloride28.02μg/mL, which was closed to the theoretical value (30.56ug/mL).The results of anti-inflammatory and analgesic evaluation showed that:VA, VF has a better anti-inflammatory and analgesic effect than VAFS prescriptions; V has a good analgesic effect, but the anti-inflammatory effect of it was poor. Compared with the HuangDaoyi oil, HLYA efficacy is inferior, but there’s no statistically significant between the samples. The formulations of the medicated oil may be optimized by adding moderate turpentine and oleum omnamomi.