复康健元方HPLC指纹图谱和含量测定方法建立及其应用形式的研究

HPLC Fingerprinting,Multi-component Determination,and Application of Fukang Jianyuan Prescription

目的:建立具有促进奶牛产后恢复功能的复康健元方指纹图谱及多元成分定量分析的质量评价方法,探究压制成颗粒饲料对复康健元方中多元成分的影响。方法:采用高效液相色谱法-光电二极管阵列检测器(HPLC-PDA)构建复康健元方指纹图谱,同时对复康健元方和含复康健元方的颗粒中柚皮苷、新橙皮苷、芍药苷、藁本内酯、芍药内酯苷、甘草酸、绿原酸、6-姜酚和甘草苷进行定量分析。结果:复康健元方的HPLC指纹图谱共有18个色谱峰,并指认18个峰的归属,10批样品相似度均大于0.984;经对照品指认9个峰并进行定量分析,这9个成分在一定质量浓度范围内与峰面积呈良好的线性关系,r均大于0.9950,方法学考察表明,各项指标均符合要求;定量分析表明,复康健元方中9个成分含量由高到低分别为柚皮苷、新橙皮苷、芍药苷、藁本内酯、芍药内酯苷、甘草酸、绿原酸、6-姜酚和甘草苷,其中柚皮苷质量分数最高,为3191.21~4756.10μg·g^(–1);压制成颗粒饲料使6-姜酚、绿原酸、藁本内酯、甘草酸和甘草苷的含量显著降低(P<0.05),其中6-姜酚降低56.77%,对芍药苷、芍药内酯苷、柚皮苷和新橙皮苷的含量影响差异无统计学意义。结论:所建立的复康健元方HPLC指纹图谱分析方法及多指标成分含量测定方法稳定可行、专属性强,可用于其质量控制,为混合型天然植物饲料原料的质量评价和应用形式提供借鉴。

Objective:To establish a method for the quality evaluation of Fukang Jianyuan(FKJY)Prescription with the function of promoting postpartum recovery of cows based on fingerprinting and quantitative analysis of multiple components and explore the influences of pellet feed production by pressing on the multiple components.Methods:High performance liquid chromatography-photo-diode array(HPLC-PDA)was used for the fingerprinting of FKJY Prescription.At the same time,nine components such as chlorogenic acid in FKJY Prescription and the pellet feed containing FKJY were quantitatively analyzed by HPLC-PDA.Results:The HPLC fingerprint of FKJY Prescription was established.It presented 18 common peaks,which were identified as 18 components.The fingerprints of 10 batches of FKJY Prescription samples showed the similarity above 0.984.The content of nine components was determined.All of the calibration curves showed good linear regression(r>0.9950)within the tested ranges.The methodological investigation results showed that all indicators met the requirements.The quantitative results showed that the content of 9 compounds in FKJY Prescription was in a descending order of naringin,neohesperidin,paeoniflorin,ligustilide,paeoniflorin,glycyrrhizic acid,chlorogenic acid,6-gingerol,and glycyrrhizin,among which naringin had the highest content of 3191.21-4756.10μg·g-1.The content of 6-gingerol,chlorogenic acid,ligustilide,glycyrrhizic acid,and glycyrrhizin decreased significantly in the process of pressing pellet feed(P<0.05),among which 6-gingerol decreased by 56.77%.The pellet feed production process had no significant effect on the content of paeoniflorin,albiflorin,naringin,and neohesperidin.Conclusion:The HPLC fingerprinting and multi-component determination method was stable,feasible,and specific and it can be used for the quality control of FKJY Prescription.The findings provide reference for the quality evaluation and application of raw materials of plant-derived feed.