Impacts of Different Procedures on the Extraction of Total Flavonoids from Chickpea(Cicer arietinum)and the Analgesic Effect on Mice
[Objective]This study aimed to determine the optimal extraction and processing method for total flavonoids from chickpea(Cicer arietinum),and to evaluate the analgesic effect of chickpea total flavonoids(CTFs)on mice.[Method]The chickpeas produced in Xinjiang Uygur Autonomous Region were used as the experimental materials.The aluminum nitrate-sodium nitrite colorimetric method was employed to assess the contents of CTFs obtained through ultrasonic-assisted extraction,Soxhlet reflux extraction,micro-wave extraction,ultrasonic-assisted extraction+rotary evaporation treatment,Soxhlet reflux extraction+rotary evaporation treatment,ultrasonic-assisted extraction+freeze-drying treatment,respectively,to determine the optimal extraction and processing method for CTFs.Kunming mice were randomly assigned into 5 groups.The high,medium and low dose groups were gavaged with 60,30 and 15 mg/(kg·BW)of CTFs.The positive control group was gavaged with 80 mg/(kg·BW)of aspirin.The blank control group was gavaged with 0.1 mL/(10 g·BW)of physiological saline.Each group was administered once daily for 3 consecutive days.According to the above grouping and administration scheme,the analgesic effects of different doses of CTFs on mice were evaluated using hot plate test(n=40,8 mice in each group),warm bath test(n=40,8 mice in each group)and acetic acid twisting test(n=40,8 mice in each group).[Result]The CTFs obtained by different extraction and processing methods were as follows:ultrasonic-assisted extraction+freeze-drying treatment(51.18 mg/g)>Soxhlet reflux extraction(13.16 mg/g)>ultrasonic-assisted extraction(4.86 mg/g)>Soxhlet reflux extraction+rotary evaporation treatment(4.75 mg/g)>micro-wave extraction(3.03 mg/g)>ultrasonic-assisted extraction +rotary evaporation treatment(0.57 mg/g).Hot plate test demonstrated that compared with the blank control group,CTFs high and low dose groups had extremely significantly(P<0.01)prolonged pain threshold at 10,30 and 60 min after administration,while CTFs medium dose group exhibited significantly(P<0.05)prolonged pain threshold at 10 and 30 min after administration.CTFs medium and low dose groups had extremely significantly(P<0.01)prolonged pain threshold in warm bath test at 2 h after administration than the blank control group.CTFs high,medium and low dose groups had extremely significantly(P<0.01)reduced the frequency of twisting induced by acetic acid compared with the blank control group,with the pain inhibition rates of 35.44%,35.44%and 61.37%,respectively.In addition,the frequency of twisting in CTFs low dose group was lower than that in the positive control group,with higher pain inhibition rate than that in the positive control group.[Conclusion]The optimal extraction and processing method for CTFs was ultrasound-assisted extraction+freeze-drying treatment.Different doses of CTFs had good analgesic effects on mice,with low dose[15 mg/(kg·BW)]having the best.
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