Chemical Components Analysis And Quality Control Of Wanglaoji Herbal Tea

Objective:Wanglaoji Herbal Tea is famous as one of the Cantonese Herbal Tea which possesses actions to eliminate heat and toxic materials,cure thirst disease,bases on the theory of TCM(Traditional Chinese Medicine).It is consisted of seven Chinese herbal medicines as Chinese Mesona,Mexican Fangipani,Lonicerae Japonicae Flos,Chrysanthemi Flos,Prunellae Spica,Microctis Folium and Glycyrrhizae Radix et Rhizoma.Chinese medicine contains kinds of chemical compositions,so as Wanglaoji Herbal Tea which is consisted of seven kinds of Chinese medicine.At present,there are some deficiencies in the quality evaluation for Wanglaoji Herbal Tea in industry,such as the lack of specificity.In order to ensure the stability of Wanglaoji Herbal Tea preferably,it is necessary to carry out the analysis for the ingredients of main effective components.Therefore,we took the two kinds of samples from two procudures as objects,established a simple and specificity method for the identification and quality evaluation of Wanglaoji Herbal Tea effectively,which provided the basis for the upgrading of Wanglaoji Herbal Tea products.Method:1.The HPLC separation was performed on a ZORBAX SB-C18(150 × 4.6mm,1.8?m)by using acetonitrile and 0.1%formic acid aqueous solution as mobile phase with the gradient elution.To find out the different components between two kinds of processes'samples,and monitored the transmission from semi-finished products to finished products in the negative ion mode.2.The GC-MS method was applied with a fiber colum HP-5 MS(30.0 m × 250 ?m,0.25 ?m)in temperature programmer,with He as the carrier gas.Calculated the relative content of each compound by area normalization method,and compared the different volatile components of Wanglaoji Herbal Tea products from two processes.3.Established the fingerprint of Wanglaoji herbal tea products from BM process and ST process by HPLC,the separation was performed on a Phenomenex Luna C18(250 × 4.6 mm,5 ?m)colum by using acetonitrile and 0.1%formic acid aqueous solution as mobile phase with the gradient elution,the column temperature was 20 ?.The detection wavelength was 235 nm.The fingerprints were compared and comprehensively evaluated by the evaluation system of Traditional Chinese Medicine fingerprint and statistical software.4.The GC analysis was performed on DB-WAX(30 m × 0.25 mm,0.25 ?m)colum,with FID detector,nitrogen was used as the carrier gas with its flow rate 3.0 mL/min.The flow rate of atmosphere was 400 mL/min,and the flow rate of hydrogen was 40.0 mL/min in temperature programer.The fingerprint was comprehensively evaluated by the evaluation system of Traditional Chinese Medicine fingerprint.5.Based on the HPLC fingerprint,we analyzed the contents of tanshinol,cerberic acid B,15-demethylplumieride,rosmarinic acid glycosides and glycyrrhizic acid.Then proposed the content limit of four main target components included tanshinol,cerberic acid B.15-demethylplumieride and glycyrrhizic acid.6.To develop a method for the determination of disacoside B,macranthoidin B,caffeoylplumieride,narcissoside,rutin,keampferol-3-O-?-D-glucose-7-O-a-L-rhamnase,isochlorogenic acid A,trifolin,liquiritin,rosmarinic acid,chlorogenic acid,caffeic acid,protocatechuic acid and protocatechualdehyde in Wanglaoji Herbal Tea by HPLC/MS/MS.The HPLC separation was performed on an Aglient Poroshell 120 EC-C18(50 × 3.0mm,2.7?m)by using acetonitrile and 0.1%formic acid aqueous solution as mobile phase with the gradient elution.The column temperature was 25 ?.The analytes were detected in negative mode by tandem mass apectrometry with the ESI source and carried out under the multiple reaction monitoring(MRM)modes.To find out the main target components by statistical analysis method.Results:1.The UHPLC-MS analysis results presented that the key chemical constituents of Wanglaoji Herbal Tea were phenolic acids and flavonoids.As 29 compounds were isolated in semi-products of ST process,among which phenolic acids(11 kinds)had the most types and flavonoids(6 kinds)followed.And 22 compounds were isolated in finished-products of ST process which contained 9 kinds of phenolic acids and 4 kinds of flavonoids followed.Simultaneously,37 compounds were isolated in semi-products of BM process contained 14 kinds of phenolic acids and 7 kinds of flavonoids followed,29 compounds were isolated in finished-products of BM process contained 13 kinds of phenolic acids and 4 kinds of flavonoids followed.The information showed that BM process extracted relatively completely,and the chemical composition of semi-finished products almostly contained the products of finished.Therefore,the transfer rate of the BM process from the intermediate to the finished product is 73.0%as 72.4%in ST process,which showed that the chemical compositions were transmitted well in these two processes.2.The GC-MS analysis presented that scarcely volatile substance in the end products of Wanglaoji Herbal Tea.However,46 compounds were isolated and discerned in semi-products of BM process among which the alcohol as the key constitiuent accounted for 15.0%.So as semi-finished products of ST process can be separated to 25 components contained the mainly ingredient furane accounted for 17.0%.The results exposed that BM process could extract the volatile components completely.3.The fingerprint similarity of Wanglaoji Herbal Tea from two kinds of processes was higher than 0.95,which was created by HPLC-DAD method.Indicating these two processes were stability.The cluster analysis divided 40 batches into 4 groups,the same process samples gathered for a class,samples of the same process under different product phases(semi-finished products,finished products)respectively gathered as a class,which is the same as the actual classification.4.The GC fingerprint similarity about products of BM process was higher than 0.98,reflecting that this production process was steady.5.The determination of 5 chemical components by HPLC indicated that components of 15-demethylplumieride and cerberic acid B had higher extraction in BM process.Tanshinol,rosmarinic acid glycosides and glycyrrhizic acid were higher in ST process'samples.In addition,according to the content of 4 taget components,we suggesed that tanshinol was not less than 4.9 ?g/mL,cerberic acid B was not less than 6.5 ?g/mL,15-demethylplumieride was not less than 2.8 ?g/mL and glycyrrhizic acid was not less than 8.0 pg/mL in BM process finished products.Concurrently,tanshinol was not less than 13.7?g/mL,cerberic acid B was not less than 2.8 ?g/mL,15-demethylplumieride was not less than 1.8 ?g/mL and glycyrrhizic acid was not less than 14.2 ?g/mL in ST process finished products.6.The determination of 14 chemical components in Wanglaoji Herbal Tea by HPLC/MS/MS demonstrated that 6 components of keampferol-3-O-?-D-glucose-7-O-?-L-rhamnase,rutin,coffeic acid,trifolin,chlorogenic acid,and caffeoylplumieride had higher extraction in BM process.And 5 components of disacoside B,macranthoidin B,liquiritin,rosmarinic acid,protocatechualdehyde were higher in ST process' samples.Additionally,narcissoside,isochlorogenic acid A and protocatechuic acid were equal in these two processes.According to the principal component analysis we known the 5 components of liquiritin,rosmarinic acid,caffeic acid,narcissoside,and protocatechualdehyde were significantly important in the classification of Wanglaoji hebal tea.In hence,we suggesed that liquiritin was not less than 1.5 ?g/mL,rosmarinic acid was not less than 6.4 ?g/mL,caffeic acid was not less than 1.9 ?g/mL,narcissoside was not less than 1.0 ?g/mL and protocatechualdehyde was not less than 0.4 ?g/mL in BM process finished products.Concurrently,liquiritin was not less than 2.6 ?g/mL,rosmarinic acid was not less than 7.6?g/mL,caffeic acid was not less than 0.4 ?g/mL,narcissoside was not less than 1.0 ?g/mL and protocatechualdehyde was not less than 0.5 ?g/mL in ST process finished products.Conclusion:1.The established HPLC-MS and GC-MS qualitative analysis method comprehensively reflected the information of volatile and non-volatile components in Wanglaoji Herbal Tea,laying the foundation for the further establishment of the quality evaluation system of Wanglaoji Herbal Tea.2.The established fingerprint method would distinguish the different processes of Wanglaoji Herbal Tea,and reflected the overall characteristics and individual characteristics of two processes's products,intuitively and quickly identify the Wanglaoji Herbal Tea.3.The determination for the content of multi-index components generally reflected the condition about the seven raw materials,which can be used to control the quality of Wanglaoji Herbal Tea.In this paper,we established a quality control mode which contained fingerprint and determination of multiple components.Observed the raw material,semi-finished products and finished products,the major components had a perfectly correlation and strongly ascripition,which comprehensively reflected the overall quality of Wanglaoji Herbal Tea.