不同生长年限鲜参与生晒参多类型化学成分差异分析与评价

Differential analysis and evaluation of multiple types of chemical components in fresh and white ginseng with different growth years

目的:探讨不同生长年限鲜参与生晒参中多类型成分的差异,为人参质量控制及开发利用提供科学依据。方法:采用高效液相色谱-蒸发光散射检测器法(HPLC-ELSD法)对人参中皂苷类化学成分组成与含量进行分析;分析条件:采用Dimaonsil®ODS C_(18)(250 mm×4.6 mm,5μm)色谱柱,以乙腈(A)-水(B)为流动相,梯度洗脱,流速1.0 mL·min^(-1),蒸发光散射检测器漂移管温度100℃,气体流量2.8 L·min^(-1)。采用紫外-可见分光光度法测定人参中可溶性多糖含量,以葡萄糖与葡萄糖醛酸为对照品测定中性多糖与酸性多糖含量,检测波长分别为490、512 nm。采用超高效液相色谱-三重四极杆质谱串联法(UPLC-T Q MS法)对人参中氨基酸类与核苷类化学成分组成与含量进行分析;分析条件:采用ACQUITY UPLC BEH Amide(100 mm×2.1 mm,1.7μm)色谱柱,以含有5 mmol·L^(-1)甲酸铵、5 mmol·L^(-1)乙酸铵和0.2%甲酸的水溶液为流动相A,以含有1 mmol·L^(-1)甲酸铵、1 mmol·L^(-1)乙酸铵和0.2%甲酸的乙腈溶液为流动相B,梯度洗脱,流速0.40 mL·min^(-1),柱温为30℃,进样量为2μL;电喷雾(ESI)离子源,正离子模式多反应监测采集。结果:在相同生长年限下,生晒参中8个皂苷类成分(人参皂苷Re、人参皂苷Rg1、人参皂苷Rf、人参皂苷Rb1、人参皂苷Rc、人参皂苷Rb2、人参皂苷Rb3、人参皂苷Rd)与7个核苷类成分(胸腺嘧啶、胸苷、尿苷、腺苷、胞苷、鸟苷、腺嘌呤)平均总量分别为7.10~12.75、0.1949~0.8784 mg·g^(-1),均高于鲜参;鲜参中可溶性多糖(中性多糖与酸性多糖)与15个氨基酸类成分(L-亮氨酸、L-苯丙氨酸、L-色氨酸、γ-氨基丁酸、L-异亮氨酸、L-缬氨酸、L-脯氨酸、L-酪氨酸、β-丙氨酸、L-苏氨酸、L-谷氨酰胺、L-天冬酰胺、L-天冬氨酸、L-精氨酸、L-赖氨酸)平均总量分别为11.03%~18.29%、7.51~13.58 mg·g^(-1),均高于生晒参。比较3~6年生鲜参与生晒参,发现在6年生人参中可溶性多糖、8个人参皂苷类成分、15个氨基酸类成分与7个核苷类成分的平均总量最高,分别为18.29%、12.75 mg·g^(-1)、13.58 mg·g^(-1)、0.8784 mg·g^(-1)。结论:不同生长年限鲜参与生晒参多类型成分含量具有差异性,且随生长年限的延长,其可溶性多糖、8个人参皂苷类成分、15个氨基酸类成分与7个核苷类成分总量呈增加趋势。研究结果为鲜参与生晒参药效差异物质基础与质量控制提供科学依据。

Objective:To explore the differences of multiple types of chemical constituents in fresh and white ginseng with different growth years,which provided reference for the quality control and comprehensive exploitation of Panax ginseng.Methods:The saponins in ginseng was determined by HPLC-ELSD;Analytical conditions:a Dimaonsil®ODS C_(18)(250 mm×4.6 mm,5μm)column was used with(A)-water(B)(gradient elution)as the mobile phase at a flow rate of 1.0 mL·min^(-1),the temperature of the drift tube was 100℃,the gas flow rate was 2.8 L·min^(-1).The UV-Vis spectrophotometric was used to determine the soluble polysaccharides.Glucose and glucuronic acid were used as reference substances of the neutral polysaccharide and acidic polysaccharide with detection wavelengths of 490 nm and 512 nm,respectively.UPLC-T Q MS was used for analyzing amino acids and nucleosides of Panax ginseng.Analytical conditions:an ACQUITY UPLC BEH Amide(100 mm×2.1 mm,1.7μm)column was used with an aqueous solution containing 5 mmol·L^(-1)ammonium formate,5 mmol·L^(-1)ammonium acetate,and 0.2%formic acid as mobile phase A,and an acetonitrile solution containing 1 mmol·L^(-1)ammonium formate,1 mmol·L^(-1)ammonium acetate,and 0.2%formic acid as mobile phase B with gradient elution at the flow rate of 0.40 mL·min^(-1).Column temperature was 30℃,and injectionvolume was 2μL.Electrospray ion source was adopted with positive ion modes and multi-reaction monitoring and acquisition.Results:Under the same growth years,the content of 8 ginsenosides(ginsengside Re,ginsengside Rg_(1),ginsengside Rf,ginsengside Rb_(1),ginsengside Rc,ginsengside Rb_(2),ginsengside Rb_(3),ginsengside Rd)and 7 nucleosides(thymine,thymidine,uridine,adenosine,cytidine,guanosine,adenine)in white ginseng were higher than that in fresh ginseng,with the average content of 7.10-12.75 mg·g^(-1)and 0.1950-0.8784 mg·g^(-1),respectively.The soluble polysaccharides(neutral polysaccharide,acid polysaccharide)and 15 amino acids(L-leucine,L-phenylalanine,L-tryptophan,gamma-aminobutyric,L-isoleucine,L-valine,L-proline,L-tyrosine,β-alanine,L-threonine,L-glutamine,L-asparagine,L-asparticacid,L-arginine,L-lysine)in fresh ginseng were higher than that in white ginseng,with the average contents of 11.03%-18.29%and 7.61-13.58 mg·g^(-1),respectively.Comparison of 3-6 years old with fresh ginseng and white ginseng revealed that the highest average total amounts of soluble polysaccharides,8 ginsenosides,15 amino acids and 7 nucleosides were found in the 6 years old ginseng,which were 18.29%,12.75 mg·g^(-1),13.58 mg·g^(-1),0.8784 mg·g^(-1).Conclusion:The contents of multiple types of components in fresh ginseng and white ginseng from different growth years were different.The total amount of soluble polysaccharides,8 ginsenosides,15 amino acids and 7 nucleosides all increased with growth years.The results provide scientific basis for efficacy differences and the quality control of fresh ginseng and white ginseng.