杨畅++白洁++杜守颖++崔雅华++张勤帅++马军明

[摘要] 建立活血止痛膏中有效成分丹皮酚、丁香酚、胡椒碱微透析体内外回收率校正方法，为其经皮药动学研究提供依据。利用高效液相色谱法测定透析液中丹皮酚、丁香酚、胡椒碱的浓度，并计算微透析探针回收率；研究浓度、校正方法对3种成分微透析探针回收率的影响，并考察探针吸附性、体内外回收率的稳定性和重复性。结果表明，以30%乙醇生理盐水作为灌流液时，探针对3种成分均无明显的吸附，短时间内即可从探针上被完全洗脱下来；在8 h 内，3种成分体内外回收率均能保持稳定；交替用含药溶液和空白灌流液灌流探针3次，3种成分体内外回收率基本不变。不同质量浓度丹皮酚（2.46～53.64 mg·L-1）、丁香酚（4.26～32.15 mg·L-1）和胡椒碱（5.75～45.87 mg·L-1）体外回收率分别为（45.7±4.66）%，（41.3±3.96）%，（27.82±2.95）%，即3种成分的探针回收率对浓度没有依赖性；在相同条件下，零净通量法、透析法和反透析法所测得的探针体外回收率几乎相等。因此，采集部位药物的浓度可用体外测得的回收率或在体回收率进行校正。同时也证明了该研究建立的微透析方法可用于活血止痛膏的经皮药动学研究。

[关键词] 微透析； 探针回收率； 丹皮酚； 胡椒碱； 丁香酚

In vitro and in vivo recoveries of cutaneous microdialysis probe of paeonol， eugenol and piperine

YANG Chang， BAI Jie*， DU Shouying*， CUI Yahua， Zhang Qinshuai， Ma Junming

（Beijing University of Chinese Medicine， Beijing 100102， China）

[Abstract] To establish a method for detecting microdialysis recovery of paeonol， eugenol and piperine in Huoxue Zhitong patch， in order to provide the basis for further percutaneous pharmacokinetics studies. The concentrations of paeonol， eugenol and piperine in dialysates were determined by HPLC， and probe deliveries were calculated respectively. The effects of concentration and calibration approaches on the microdialysis probe deliveries of the three components were investigated， and their probe absorbability， in vitro and in vivo probe stability and repeatability were also studied.The results indicated that little paeonol， eugenol and piperine were observed in probes with 30% alcohol as the perfusate， and could be cleaned from probe in a short time. And the in vivo and in vitro probe deliveries of three components were stable within 8 h， drugcontaining solution and blank perfusate were alternatively used for three times， and the in vivo and in vitro probe deliveries of three components were basically unchanged. The in vitro recoveries of paeonol， eugenol and piperine with a range of concentration were respectively （45.7±4.66）%， （27.82±2.95）%， （41.3±3.96）%， which indicated no concentration independent. Under the same conditions， the similar delivery was observed by dialysis， retrodialysis and nonet flux. Therefore， the concentrations of analyses of the collected fraction could be calibrated by in vitro or in vivo recoveries. Meanwhile， this also proved that the microdialysis method built by this study is applicable to the study on percutaneous pharmacokinetics of Huoxue Zhitong patch.

[Key words] microdialysis； probe delivery； paeonol； eugenol； piperine

doi：10.4268/cjcmm20162226

活血止痛膏应用历史悠久，能够活血止痛、舒筋通络，临床上广泛用于筋骨疼痛、肌肉麻痹、痰核流注、关节疼痛等的治疗。其中牡丹皮Paeonia suffruticosa Andr.为方中君药，与当归、川芎等配伍能够活血化瘀止痛，胡椒Piper nigrwm L.，丁香Eugenia caryophyllata Thunb.等辛温之品，加强散寒止痛之功。丹皮酚（paeonol，Pae）、丁香酚（eugenol，Eug）和胡椒碱（piperine，Pip）分别是牡丹皮、丁香和胡椒的主要活性成分，均为活血止痛膏中的有效成分[1]。……