基于微量量热法的连花清瘟胶囊质量一致性评价方法的建立

目的考察连花清瘟胶囊(LQC)对绿脓杆菌Pseudomonas aeruginosa生长代谢的影响,建立评价LQC质量一致性的新方法。方法采用微量量热法测定不同质量浓度LQC对绿脓杆菌的生长代谢的影响,得到热谱曲线及相应的热动力学参数,如生长峰达峰时间(T)、生长期发热量(Q)、生长期最大发热功率(P)、生长期生长速率常数(k)等。以LQC质量浓度(C)为横坐标,T、P、Q、k为纵坐标进行回归分析,根据相关系数优选最佳参数,计算抑菌有效率(antibacterial effective rate,E)。以E作为评价LQC抗菌活性指标,分别检测11个批次市售LQC以及经特殊处理后的LQC样品对绿脓杆菌生长代谢的影响。结果 LQC质量浓度在10.00~47.68 mg/m L时对绿脓杆菌生长代谢具有明显的抑制作用,且具有质量浓度依赖性。通过回归分析得出的4个热动力学参数中,相关性最好的参数为T,r=0.990,其变化可以直观评价LQC对绿脓杆菌生长代谢的影响,且该参数稳定性较好,故选择指数生长期的T计算E。结果表明不同批次LQC样品E无明显差异,样品放于潮湿或高温环境中其E发生明显变化。结论该方法具有灵敏度高、重现性好的特点,可以区分不同质量LQC的抗菌生物活性,为LQC质量的一致性评价提供了一种参考方法,并为其他大复方中药制剂质量评价提供了参考依据和方法学借鉴。

Consistency evaluation method for Lianhua Qingwen Capsule based on microcalorimetry

Objective To study the effect of Lianhua Qingwen Capsule (LQC) on the growth and metabolism of Pseudomonas aeruginosa and to establish a new method to evaluate the consistency of LQC. Methods The power-time curves and corresponding thermodynamic parameters such as appearance time (T), total heat output (Q), maximum power output (P), and growth rate constant (k) of P. aeruginosa were determined by microcalorimetry. The regression analysis was carried out with T, P, Q, and k as parameters, and the optimal parameters were selected according to the correlation coefficient to calculate the antibacterial effective rate. The effects of 11 batches of LQC and specially treated LQC samples on growth and metabolism of P. aeruginosa were determined by an index antibacterial effective rate to evaluate the antibacterial activity of LQC. Results LQC had obvious inhibitory effect on P. aeruginosa in the range of 10.00 to 47.68 mg/mL, and the bacteriostatic efficacy and dosage of LQC had good correlation. By regression analysis, it was found that T was the best parameter in the four thermodynamic parameters, r=0.990. The change of this parameter could directly evaluate the effect of LQC on the growth and metabolism of P. aeruginosa, with good stability. The appearance time was selected as a parameter to calculate its antibacterial effective rate (E). The results showed that there was no significant difference in E of different batches of LQC samples. And the E value was significantly changed when the sample was placed in a humid/high temperature environment. Conclusion The microcalorimetric method can be used to evaluate the antibacterial activity and the quality consistency of LQC capsule, which has the high precision and good accuracy. The study has provided a new method to evaluate the quality consistency of TCM.