Establishment And Application Of HPLC Fingerprint Before And After Fermentation Of Piweisan

Piweisan uses Codonopsis Radix as the main monarch to tonify the middle Jiao,strengthen the spleen and benefit the lung,and play the main therapeutic role for the main evidence.Fried Atractylodis Macrocephalae Rhizoma and Poria are the complementary subjects,which together have the effect of strengthening the spleen and inducing diuresis,in order to supplement and enhance the spleen-strengthening function of Codonopsis Radix.Citri Fructus,Fructus Noveniae,Aurantii Fructus Immaturus,Hippophae Fructus,Ginger Processed Pinellia,Wume Fructus,Citrus Aurantium,Citri Sarcodactylis Fructus and Perilla Frutescens all adjuvar,which have the effects of soothing the liver and Qi,dispelling phlegm and relieving cough,and harmonizing the stomach and resolving stagnation,in order to alleviate the secondary symptoms and treat the multiple symptoms of stomach diseases.Glycyrrhiza Uralensis is used to harmonize all the medicines,to relieve the toxic potency of the medicines in this formula,and to increase their synergistic effect.Piweisan can be used for the treatment of chronic gastritis,and the clinical use of this herbal compound has proved its good efficacy,but there is a lack of suitable quality standards for Piweisan.In this paper,we used Aspergillus oryzae as the fermenting strain for the solidstate fermentation of Piweisan as the solid-state medium for the concoction of Piweisan.Firstly,the active ingredients of Piweisan fermentation were investigated by UPLC-MS and HPLC,based on which the solid-state fermentation process of Piweisan was optimized using response surface methodology and the quality control standard of Piweisan was established.The conclusions were as follows:(1)The components of Piweisan were analyzed by UPLC-MS,and 102 substances were detected,among which flavonoids accounted for the highest proportion of active ingredients(37).Flavonoids have the effects of accelerating gastric mucosa repair,progastrointestinal motility,anti-inflammation and antioxidation,and can be used as indicator components of Piweisan;(2)The HPLC fingerprints of Piweisan were established,and a total of 48 common peaks were identified,9 of which were attributed and quantitatively analyzed at 1.36mg/g for Siniferin,0.25 mg/g for Adenosine,0.34 mg/g for Nicotinic acid,0.29 mg/g for Dihydromyricetin,0.80 mg/g for Glycyrrhizin,1.38 mg/g for Naringenin,2.29mg/g for Hesperidin,4.64 mg/g for Neohesperidin,0.45 mg/g for Rosmarinic acid,the similarities of the established fingerprint profiles were all greater than 0.9,which could be used as quality control standards for Piweisan;(3)After response surface optimization experiments,the optimal fermentation conditions for the fermentation concoction of Piweisan were confirmed as follows: 65%water content,4.5 d fermentation time,6.5% inoculum,and the average reduction of peak area of the common peaks of Piweisan measured under these conditions was90.36%,which was in accordance with the expected results;(4)The HPLC fingerprints of the solid-state fermentation concoction of Piweisan were established,25 shared peaks were identified,7 of them were attributed and quantitatively analyzed,and the contents of the active ingredients were 1.92 mg/g of Siniferin,0.38 mg/g of Adenosine,0.23 mg/g of Nicotinic acid,0.02 mg/g of Dihydromyricetin,0.03 mg/g of Naringenin,0.63 mg/g of Neohesperidin and 0.10 mg/g of Neohesperidin,the similarity of the established fingerprint profiles were all greater than 0.9,which can be used as quality control standards for the solid state concoction of Piweisan;(5)The HPLC chromatograms of 10 batches of fermentation tank concoction of Piweisan and the control HPLC fingerprint profiles of solid-state fermentation concoction of Piweisan had similarities greater than 0.9,which met the established quality control standards of fermentation concoction of Piweisan;(6)Solid-state fermented concoction of Piweisan has better clinical therapeutic effect on patients with Chronic gastritis,and the therapeutic efficiency of fermented concoction of Piweisan reached 97.14%,which was higher than that of unfermented Piweisan group(89.89%).