A Study On Preparation Process And Quality Standard Of Yifa Tincture

Yifa Tincture,formerly known as Shengfaling Tincture,was a Chinese herbal compound preparation for external use researched and developed by Nanfang hospital. It was composed of seven Chinese herbs,such as Radix Paeonia rubra,Radix Angelicae sinensis,Flos Carthami,Cacumen Biotae,and prepared by volatile oil extraction and 75%ethanol impregnation processes.Yifa Tincture was indicated for alopecia areata caused by hemopenia and blood stagnation.The clinical observation showed that the total efficiency of treatment group was 93.9%in 280 cases of alopecia areata patients and was better than that of control group significantly.On the basis of the clinical observation,the blood theology and the hair grows of the model rats with acute blood stasis,hair removal,qi-deficiency were investigated after the model rats were treated with Yifa Tincture.The results showed that Yifa Tincture had the effects of activating blood circulation to dissipate blood stasis and could promote the hair growth.And then,the preliminary quality control of the preparation was established by double-wave TLC scanning method.Because of the coarse preparation process and the backward quality standard,the clinical effects of the tincture were influenced.Therefore,it was necessary to improve the preparation process and quality standard. Objectives:The preparation process and quality control of Yifa Tincture were studied systematically to screen a reasonable and economical preparation process,and establish an accurate,reproducible and specific quality standard.It can not only serve better for clinic,but also provide the experimental basis for the further development of Yifa Tincture.Methods:1.To optimize the extraction process of Yifa Tincture1.1 All of the herbs and the herbs contained violate oil were extracted respectively by water-steam distillation,and the yield of volatile oil was investigated.1.2 The influence factors of extraction procedure,the concentration of alcohol,the volume of alcohol,and the extraction time,were considered by the means of orthogonal design indicated by the content of ferulaic acid.2.To establish the quality standard of Yifa Tincture2.1 The main herbs of the tincture were respectively identified by TLC referred to the reference substances and reference herbs.2.2 The content of paeoniflorin and ferulic acid in Yifa Tincture was determined by HPLC,and the methodology was also investigated.Results:1.Extraction technology of Yifa Tincture1.1 The yield of volatile oil had no significant difference between the two methods. The amount of volatile oil did not increase after the herbs were extracted for 4.5 h. Thus,the optimal process was defined that the herbs contained volatile oil was extracted first for 4.5h and then combined with the other herbs to lixiviate and prepare the tincture.1.2 The content of ferulaic acid amounted to the highest when the herbs were impregnated with 8 times volumes of 75%alcohol corresponding to the amount of the herbs for 120 h.The analysis of variance revealed that the concentration of alcohol and the extraction time had significant influence on the preparation process.2.Quality standard of Yifa Tincture2.1 The six herbs in the tincture had been respectively identified by TLC,and the specific chromatogram of Radix Paeonia rubra,Radix Angelicae sinensis,Flos Carthami,Cacumen Biotae,Radix Polygoni multiflori praeparata cum succo glycines sotae and Radix Rehmanniae was observed with clear dot and good resolution.The negative sample had no interference and the method was specific.2.2 The methodology investigation proved that the determination of peaoniflorin and ferulic acid by HPLC was satisfied by liner correlation,precision,reproducibility, stability and recovery.The average content of peaoniflorin and ferulic acid in Yifa Tincture was 0.0348mg/ml and 0.0375mg/ml respectively.Conclusion:The optimal preparation process was as follows:the tree herbs contained volatile oil was extracted by water-steam distillation first,and then combined with the other herbs consisted of the prescription and impregnated with 8 times volume of 75% alcohol for 120 h.Under the optimal conditions,the content of ferulic acid in Yifa Tincture was satisfied.The TLC identification of the herbs in tincture was conformed to the requirements.The determination method of peaoniflorin and ferulic acid in tincture by HPLC was simple,accurate and had a good reproducibility.The established method could be used for the control of Yifa Tincture quality.