Evaluating the Quality of Volatile Oil Extracted from Nutmeg by CO2 Supercritical Extraction by Dose-effect Relationship
Objective:Gas chromatography-triple quadrupole mass spectrometry(GC-MS/MS)was used to quantitatively analyze terpinen-4-ol,α-pinene,methyleugenol,elemicin,safrole and myristcin in 16 batches of volatile oil of nutmeg.Combined with anti-inflammatory activity dose-effect analysis,the quality of the active ingredients in nutmeg was evaluated.Methods:CO2 supercritical extraction method was used to extract the volatile oil of nutmeg,and a GC-MS/MS method was established to analyze the six components in nutmeg.Methodological investigation was conducted and the content of six components in 16 batches of volatile oil samples was determined.Gas chromatographic conditions:J&W DB-17ms gas column(30 m×0.25 mm,0.25 μm);temperature rise procedure:50 ℃,keep for 1 min,rise to 150 ℃ at the rate of 10 ℃·min-1,keep for 3 min;then it was raised to 250 ℃ at the rate of 20 ℃·min-1 and keep for 3 min.Inlet temperature 280 ℃;the carrier gas was nitrogen,the flow rate was 1 mL·min-1,the shunt ratio was 10∶1,and the sample size was 1 μL.The inflammation model of Raw264.7 macrophages induced by lipopolysaccharide(LPS)was established in 16 batches of volatile oil,and the pharmacodynamic analysis was performed.The volatile oil samples were analyzed by principal component analysis(PCA-X)and Pearson coefficient was positive compounds as contributing components.Results:There was a good linear relationship between terpinen-4-ol,α-pinene,methyleugenol,elemicin,safrole and myristicin in the corresponding concentration range.RSD of the six components ranged from 0.18％to 0.90％for precision test,1.53％to 2.90％for repeatability test,and 2.11％-2.94％for stability test.The average recoveries(n=9)were 98.32％,99.41％,94.79％,98.82％,96.14％,97.81％,and RSD was 1.51％,1.62％,1.87％,0.71％,1.82％,1.31％,respectively.The contents of terpinen-4-ol,α-pinene,methyleugenol,elemicin,safrole and myristicin in 16 batches of volatile oil samples ranged from 4.674-12.432 mg·g-1,2.316-11.121 mg·g-1,0.201-4.653 mg·g-1,1.047-10.488 mg·g-1,201.776-402.163 mg·g-1,7.888-39.570 mg·g-1,respectively.The results showed that there was a correlation between the anti-inflammatory activity and the content of six components.The anti-inflammatory activity of methyleugenol,elemicin and myristicin contributed,and elemicin contributed the most.Conclusion:A dose-effect relationship analysis was established for the anti-inflammatory activity of six components in 16 batches of volatile oil of nutmeg.The method is simple and rapid,and provides a basis for the quality control,safe and effective application of nutmeg.
volatile oil of nutmegCO2 supercritical extraction methodGC-MS/MScontent determinationdose-effect relationship
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