Study On The Spectrum Effect Of Volatile Oil From Nutmeg On Relieving Chronic Pain

In order to evaluate the spectrum-effect relationships of volatile oil from nutmeg on relieving chronic pain,the optimal extraction process of traditional steam distillation and microwave-assisted steam distillation was first optimized by orthogonal design to provide an effective way for obtaining the volatile oil from nutmeg.The optimum process for the extraction of volatile oil from nutmeg by traditional steam distillation method was as follows:the particle size of the nutmeg was 40-65 mesh,12times of water was added,soaked for 1.5 hours,and extracted for 6.0 hours;The optimum process for the extraction of volatile oil from nutmeg by microwave-assisted steam distillation was crushed to a size of 40-65 mesh,300 ml of water was added,for 1.5 hours,extracted for 2.5h,and the microwave power was 390 W.Gas chromatography-mass spectrometry（GC-MS）was used to analyze the components of volatile oil from nutmeg obtained by traditional steam distillation and microwave-assisted steam distillation,and GC-MS fingerprints of volatile oil from nutmeg were established.The volatile oil from nutmeg,which were extracted by traditional steam distillation and microwave-assisted steam distillation,were mainlyβ-terpinene,（1R）-（+）-α-pinene,β-pinene,and m-cymene.In order to obtain more differentiated volatile oil from nutmeg for spectra-l studies,the volatile oil from nutmeg extracted by the above method and supercritical CO₂ fluid extraction method were fractionated by vacuum fractionation,and the components were analyzed by GC-MS.After identification,19 kinds of different volatile oil from nutmeg were finally obtained.The chromatographic similarity evaluation system of traditional Chinese medicine（2004A version）was used to analyze the control fingerprints and GC-MS fingerprints of volatile oil from nutmeg.The similarity of the comparison between the volatile oil samples were between0.008 and 0.999.There were indeed differences between the 19 volatile oils from nutmeg,which could be used as samples for spectrum analysis.According to the fingerprint chromatogram of the volatile oil from nutmeg,26 characteristic common peaks were obtained,and the sum of the peak areas accounted for 87.00%to 99.98%of the total peak area of each sample,which could represent the sample better.This article established a model of chronic inflammatory pain by subcutaneous injection of 0.15 ml Complete Freund’s Adjuvant into the midfoot of the right hind limb of Wistar rats.132 rats（150-200 g）were randomly divided into 22 groups of 6 rats each.They were fed for one week and were divided into control groups（subjected to 0.15 ml of saline in the middle of the plantar part of the right hind limb）,model group,model+positive control group（diclofenac sodium 2.5 mg/Kg）,model+volatile oil from nutmeg group（19 groups,30 mg/kg）.The control group and model group were given an equal volume of normal saline by gavage every day.The model+positive control group was given the corresponding dose of diclofenac sodium every day by gavage.The model+nutmeg volatile oil group was given the corresponding dose of volatile oil from nutmeg by gavage every day.The main observation was survival,detection of body weight,degree of toe swelling,thermal pain threshold,HE staining of rat foot sole skin and the fifth lumbar dorsal root ganglion,NK-1R,COX-2 and substance Pet al.The volatile oil from nutmeg had no significant effect on body weight,swelling structure of DRG and unformed foot.Different volatile oils from nutmeghad alleviated the toe swelling,thermal pain,NK-1R expression,COX-2 expression and substance P release by different degree.The effect of most volatile oils from nutmeg were better than model group,which indicated that the volatile oil from nutmeg has the effect of relieving chronic pain.AHP was used to determine the weight coefficient of each efficacy index for the total drug efficacy.Combining the weight coefficient of each efficacy index for the total drug efficacy,the grey relational analysis method was used to analyze the GC-MS fingerprints of volatile oil from nutmeg with the efficacy evaluation indicators and total drug efficacy.The effect of volatile oil from nutmeg in relieving chronic pain was the result of the combination of the chemical components represented by the 26 characteristic common peaks.Although the correlations between the chemical components represented by the 26 characteristic common peaks with the the efficacy evaluation indicators and total drug efficacywere different,but all of the chemical components represented by the 26 characteristic common peaks were the active ingredients of volatile oil from nutmeg to relieve chronic pain.The top five peaks which associated with relieving chronic pain were No.6peak>No.8 peak>No.9 peak>No.5 peak>No.12 peak.