GC-MS Analysis Of Volatile Components In Guben Yanling Pills And Study On The Spectrum Effect Relationship Of Tonifying Kidney Yin

Kidney-yin deficiency syndrome is a common syndrome in traditional Chinese medicine.Patients with kidney yin deficiency often have symptoms such as xerostomia,laryngoxerosis,insomnia,and soreness in the waist and lower limbs.In modern medicine,neuroendocrine immune network has been proved to be closely related to kidney-yin deficiency syndrome.Guben Yanling Pills originated from Wanbing Huichun(Ming dynasty),has the effects of strengthening the foundation,nourishing yin and marrow.It is composed of 28 medicinal herbs,with ginseng and cypress kernel as the main drugs.Ginseng volatile oil has pharmacological effects such as anti-aging,sedation,and anti-tumor,while Bai Ziren volatile oil has sedative and hypnotic effects.The ministerial drugs such as Acorus calamus,Muxiang,and Schisandra chinensis also have rich volatile components and extensive pharmacological effects.At present,the volatile components and pharmacological effects of Guben Yanling Pills have not been studied.For the further development and utilization of Guben Yanling Pills,this study established the GC-MS fingerprints of Guben Yanling Pills and studied its pharmacological action of tonifying kidney-yin by duplicating the rat model of kidneyyin deficiency syndrome.The spectrum-effect relationship of Guben Yanling Pills was established to screen its pharmacodynamic substances of tonifying kidney-yin deficiency.The main research contents of this paper are as follows:1.Study on volatile components and GC-MS fingerprints of Guben Yanling PillsThe GC-MS fingerprints of Guben Yanling Pills were established,and the method was validated.The results showed that the precision,stability and repeatability of the method met the requirements and can be used to establish the fingerprints of Guben Yanling Pills.Through similarity retrieval in the NIST database and the Kovat’s retention index method,67 volatile components were identified.Taking β-asarone as the reference peak,42 characteristic common peaks were calibrated.The similarity between the fingerprint of 20 batches of Guben Yanling Pills and the control fingerprint is 0.672～0.954.Big honey pill and little honey pill were clearly distinguished by orthogonal partial least squares discriminant analysis,and 11 differential components were founded,6 of them were identified as followed: peak 1(hexanoic acid),2(2,5-dicarboxyfuran),3(furanylhydroxymethyl ketone),4(maltol),5(3,5-dihydroxy-2-methyl-4-pyranone),and 15(β-Asarone).2.Study on the pharmacological effect of Guben Yanling Pills on tonifying kidney-yin deficiencyKidney-yin deficiency syndrome rat model was established by intragastric hydrocortisone.Body weight,anal temperature,serum hormone levels(cortisol,thyroxine)and pathological tissue sections of testis were used as evaluation indicators,while using Liuwei Dihuang Pills,commonly used for treating kidney yin deficiency syndrome,as a positive control drug,to explore the tonifying kidney-yin effect of five batches of Guben Yanling Pills.Compared with the control group,the kidney-yin deficiency model rats appeared the phenomena of yellow hair,slower growth of body mass,high anal temperature,and the levels of serum cortisol and thyroxine were significantly increased,pathological changes such as disordered arrangement of spermatogenic cells at various levels and obvious necrosis of some seminiferous tubules were observed in testicular tissue sections,indicated that the kidney-yin deficiency rat model was established successfully.Compared with the model group,each batch of Guben Yanling Pills can significantly increase the rate of body weight growth(P<0.01 or 0.05),significantly decrease the levels of serum cortisol,thyroxine and anal temperature(P<0.01 or 0.05),and adjust the testicular tissue structure to the normal level,these results suggest that Guben Yanling Pills have an ameliorative effect on kidney-yin deficiency syndrome induced by hydrocortisone.This study provides a theoretical basis for exploring the mechanism of tonifying kidney-yin of Guben Yanling Pills.3.Study on material basis of effect of Guben Yanling Pills on tonifying kidneyyin deficiencyFirstly,the data analysis methods used in the study of spectral-effect relationship were investigated,combining the advantages and disadvantages of each method and the actual situation of this experiment,grey correlation analysis(GRA)and partial least squares regression analysis(PLSR)were selected to establish the relationships between3 pharmacodynamic indexes(body weight growth rate,cortisol,thyroxine)and 42 characteristic peak areas of GC-MS fingerprints of Guben Yanling Pills of 5 batches.The chromatogram peaks which contributed to the effect of tonifying kidney-yin deficiency were screened out.The results of GRA and PLSR showed that peak 9(cismethyl isoeugenol),peak 18(unknown),peak 21(β-costol),peak 27(unknown),peak28(β-cyclocostunolide),peak 39(ferruginol),peak 41(unknown)were positively correlated with the effect of tonifying kidney-yin deficiency,may be the material basis of tonifying kidney-yin of Guben Yanling Pills.