| Gastrodia(Tianma)is the dried tubers of Gastrodia elata Bl.,which own the effect to suppress wind and antispasmodic,clam the liver and suppress yang,dispel wind and remove obstruction in the meridians.And more for the endogenous liver wind,hyperactivity of liver-yang,apoplectic dyscinesia and rheumatic arthralgia.Modern researches indicate that Gastrodia has the role of prevention of Alzheimer's disease and cardiovascular disease.In recent years,with the "traditional Chinese medicine health hot",Tianma as a healthcare products are widely favored.Tianma decoction pieces includes Tianma slice,freeze-dried Tianma slice,Tianma powder,etc.The processing method of whole Tianma capsule is direct filling Tianma medicine powder,this research regards it as a kind of Tianma herbal pieces.Because of the complexity of processing technology,the original appearance and characteristic odor of Tianma were lost or disappeared,it is difficult to identify.And the current quality of Tianma herbal pieces research only on the sum of gastrodin and p-hydroxybenzyl alcohol,it is difficult to effectively evaluate the quality of Tianma herbal pieces.So,this article study on the quality of Tianma herbal pieces in the following four aspects of the study:First,quality evaluation of various Tianma decoction pieces by HPLC fingerprint: Tianma slice,Tianma powder,whole Tianma capsule and other different characters were collected in totally 42 samples.The HPLC-DAD-MS was applicated to determination and comparison these samples,a total of 11 chromatographic peaks were identified as the common peaks in Tianma herbal pieces fingerprint.Based on the retention time,the UV spectra and the MS,the 6 peaks of gastrodin,p-hydroxybenzyl alcohol,parishin A,parishin B,parishin C,and parishin E were confirmed.These 6 peaks of HPLC fingerprint was used as the characteristic peaks.And the fingerprint of Tianma slice,freeze-dried Tianma slice,Tianma powder and whole Tianma capsule was established,and calculate the similarity of various types of Tianma herbal pieces.There were 11 common peaks and 6 characteristic peaks in the four HPLC fingerprints,the peak shape of the characteristic peaks was consistent,and the average similarity was 0.925,0.967,0.981,0.968,respectively.This four different types of Tianma herbal pieces cannot distinguished by HPLC fingerprints.Genuine,fake and adulterated products of Tianma slice and Tianma powder can be identified by HPLC fingerprint,according to the characteristics of the peak number and peak shape.Besides the authenticity of Tianma in market can be distinguished by the HPLC fingerprint of this study.Second,determination of effective components in different Tianma decoction pieces by quantitative analysis of multi-components by single-marker: A method for determining the content of 6 kinds of active ingredients in Tianma herbal pieces was established,including gastrodin,p-hydroxybenzyl alcohol,parishin A,parishin B,parishin C,and parishin E.The contents of the active ingredients of different types of Tianma herbal pieces were compared,and the quality was evaluated.And the correlation between the components was studied by Pearson correlation analysis.(1)The relative correction factor(f)of gastrodin as internal reference substance,p-hydroxybenzyl alcohol,parishin A,parishin B,parishin C,parishin E and gastrodin were 2.113,0.879,0.814,0.801,0.654,the reproducibility of each relative correction factor was good(RSD <4.217%),the results of the quantitative analysis of multi-components by single-marker are consistent with the external standard method(correlation coefficient is 0.995-1.000).(2)For all samples,except one sample cannot meet the 2015 edition of "Chinese Pharmacopoeia" standard(gastrodin and p-hydroxybenzyl alcohol can not below 0.25%),other products are in line with the standard.In compared of the sum of 6 components in Tianma herbal pieces,Tianma powder(3.214%,n=11)> freeze-dried Tianma slice(3.059%,n=3)> whole Tianma capsules(2.808%,n=13)> Tianma slice(2.366%,n=16).(3)There was a positive or negative correlation between the 6 components showed by Pearson correlation analysis.The correlation coefficient between gastrodin and pharmacopoeia standard is 0.993(P = 0.001),indicating that gastrodin is positively correlated with the sum of gastrodin and p-hydroxybenzyl alcohol,but with p-hydroxybenzyl alcohol,balicillin A,(P <0.01),indicating that the higher the content of gastrodin,the lower the content of parishin A and parishin B.Third,optimization of processing procedure for Tianma slice and freeze-dried Tianma slice:(1)Tianma slice: In this study,Tianma herb and fresh Tianma used to process Tianma slice,and the optimum process was selected by the appearance and the content of 6 kinds of effective components in Tianma slice.The processing of Tianma slice was studied by 7 methods,respectively as cold water soaked to soft,cold water moistening to soft,70? hot water immersion 4 h,70 ? hot water immersion 2 h and,steam to soft,moistening 2 h and steam to soft,direct steaming into70%-80% soft,direct steaming into totally soft.Comparison with the total contents of 6 components in Tianma is cold water moistening to soft(3.974%)> 70 degrees hot water immersion 4h(3.346%)> 70 ? hot water immersion 2h(3.158%)> direct steaming into 70%-80% soft(2.701%)> cold water soaked to soft(2.278%)> direct steaming into totally soft(2.257%).Combined with the appearance,the best way to process Tianma slice is 70 ? hot water immersion 2 h(3.158%).Fresh Tianma was processed into 3 methods,raw Tianma steamed and sliced,sliced and steamed,as well as steamed and sliced after peeled.Comparison with the total contents of 6 components in fresh Tianma is steamed and sliced(3.56%)> sliced and steamed(3.44%)> steamed and sliced after peeled(1.83%).Combined with the appearance,the best way to process Tianma slice is steamed and sliced.In addition,the effects of different drying temperature on the processing technology of Tianma were investigated.(2)Freeze-dried Tianma slice: Two different processing techniques,fresh Tianma direct slice and freeze-drying as well as freeze-drying after steamed and sliced,were investigated.And the optimum process was selected by the appearance of Tianma slice and the content of 6 kinds effective components.Comparing the sum of the six components is freeze-drying after steamed and sliced(5.32%)> fresh Tianma direct slice and freeze-drying(2.54%).Combined with the appearance,the best way to process freeze-drying Tianma slice is freeze-drying after steamed and sliced.Fourth,study on the quality markers of Tianma: Tianma contains a variety of efficacy components such as gastrodin,p-hydroxybenzyl alcohol,parishin A,parishin B,parishin C and parishin E.To choose an effective components as a maker of Tianma,the contents of 6 active components in Tianma powder were determined at 37? in different time intervals after add pepsin and trypsin.Artificial Tianma fluid hydrolysis model was established.Dynamic changes in digestive tract transformation of gastrodin,p-hydroxybenzyl alcohol,parishin A,parishin B,parishin C and parishin E was investigated.And Tianma quality markers were determined.It was found that the active components of Tianma were not transformed in the digestive tract.In the small intestine is mainly reflected in the hydrolysis products such as gastrodin.The content of gastrodin increased with the residence time,when 10 h reached the highest in 0.604%,and no more increase.The content of parishin C in 0~12 h has been increasing continuously,when 12 h reached the highest in 0.453%.The content of parishin A reached the lowest after 12 h residence,it is 58.7% compared with the highest.Gastrodin and parishin C as the main components of Tianma,can be determined as makers by artificial Tianma fluid hydrolysis model.In addition,the advancement of quality evaluation of Tianma was reviewed to provide the basis for clinical application and development of related products. |
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