分子光谱结合高效液相色谱研究人参根不同部位的差异性
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摘要
人参作为一种贵重中草药,对其研究十分广泛。常用研究方法中高效液相色谱法的应用最多,但会对样品造成不可逆的破坏,而且分析时间长。无损分子光谱技术,如红外光谱法与近红外光谱法可弥补其不足。本文以高效液相色谱法,结合红外光谱法和近红外光谱法为手段,建立了对人参根的周皮、韧皮部及木质部不同位置的差异性研究方法。
人参根的周皮、韧皮部和木质部的红外光谱间有明显的差异,尤其是周皮它含有很高的挥发油特征峰(峰2920 cm-1和1736 cm-1),但却不含多糖特征峰(峰925.7 cm-1)。主成分分析图中周皮清晰地分为一类,韧皮部和木质部的部分点有交叉。
常规近红外光谱分析粉末样品要求载样厚度需大于1 cm,论文开发了一种性能优良的厚度为1-2 mm小体积载样装置,用于少量人参根样品的近红外光谱检测。分析结果表明人参根不同部位有明显差异,而且同一部位从芦头到须根的不同位置间还有逐渐的有规律的差异。
建立了一种稳定的高效液相色谱法分析人参皂苷含量变化的方法。对色谱图分析结果表明,人参根三个部分的化学成分明显不同。从主成分分析结果看,色谱方法与红外和近红外方法的分类结果相同。表明红外光谱和近红外光谱一定程度上可以替代高效液相色谱用于人参的差异性分析。
Ginseng as a valuable Traditional Chinese Medicine, its research is a hot field. HPLC is one of the most popular methods in the research, but it will causes irreversible damage of samples and needs long analytical time normally. Molecular spectroscopy, such as IR and NIR that can supply nondestructive analysis can offset insufficiency of HPLC. A method has been established to study differences between parts of a ginseng including epidermis, phloem and xylem using infrared spectroscopy, near-infrared spectroscopy and HPLC.
Infrared spectra of ginseng epidermis, phloem and xylem are clear different. The epidermis contains very high characteristic peaks of volatile oil (peaks 2920 cm-1 and 1736 cm-1), but do not contains characteristic peak of polysaccharide (peak 925.7 cm-1). PCA result shows that epidermis clearly separated from phloem and xylem, and some dots of phloem and xylem are mixed.
Particle size and thickness of sample loaded are important interference factors for determination of near infrared spectroscopy.200 mesh of sample granularity was chosen, according to experimental comparison. Sample thickness must be over 1 cm normally to record stable and reliable spectra. In the study, large numbers of samples from different positions of ginseng were collected each containing small volume. A small volume sample holder (1-2mm thickness sample) was developed for the measurements of ginseng samples.
The analysis results of near infrared spectra also show that the epidermis, phloem and xylem parts of ginseng root are obvious different according to the NIR spectra, even spectra measured at different positions from top to bottom of epidermis, phloem and xylem show gradually changes.
Operation conditions of HPLC analysis for ginseng samples were established and the samples from the epidermis, phloem and xylem parts were analyzed. The classification results of the three parts due to HPLC data are similar with those of IR and NIR data. The results indicate that IR and NIR spectroscopy can replace HPLC to analyze the differences between different parts of epidermis, phloem and xylem of ginseng.
人参根的周皮、韧皮部和木质部的红外光谱间有明显的差异,尤其是周皮它含有很高的挥发油特征峰(峰2920 cm-1和1736 cm-1),但却不含多糖特征峰(峰925.7 cm-1)。主成分分析图中周皮清晰地分为一类,韧皮部和木质部的部分点有交叉。
常规近红外光谱分析粉末样品要求载样厚度需大于1 cm,论文开发了一种性能优良的厚度为1-2 mm小体积载样装置,用于少量人参根样品的近红外光谱检测。分析结果表明人参根不同部位有明显差异,而且同一部位从芦头到须根的不同位置间还有逐渐的有规律的差异。
建立了一种稳定的高效液相色谱法分析人参皂苷含量变化的方法。对色谱图分析结果表明,人参根三个部分的化学成分明显不同。从主成分分析结果看,色谱方法与红外和近红外方法的分类结果相同。表明红外光谱和近红外光谱一定程度上可以替代高效液相色谱用于人参的差异性分析。
Ginseng as a valuable Traditional Chinese Medicine, its research is a hot field. HPLC is one of the most popular methods in the research, but it will causes irreversible damage of samples and needs long analytical time normally. Molecular spectroscopy, such as IR and NIR that can supply nondestructive analysis can offset insufficiency of HPLC. A method has been established to study differences between parts of a ginseng including epidermis, phloem and xylem using infrared spectroscopy, near-infrared spectroscopy and HPLC.
Infrared spectra of ginseng epidermis, phloem and xylem are clear different. The epidermis contains very high characteristic peaks of volatile oil (peaks 2920 cm-1 and 1736 cm-1), but do not contains characteristic peak of polysaccharide (peak 925.7 cm-1). PCA result shows that epidermis clearly separated from phloem and xylem, and some dots of phloem and xylem are mixed.
Particle size and thickness of sample loaded are important interference factors for determination of near infrared spectroscopy.200 mesh of sample granularity was chosen, according to experimental comparison. Sample thickness must be over 1 cm normally to record stable and reliable spectra. In the study, large numbers of samples from different positions of ginseng were collected each containing small volume. A small volume sample holder (1-2mm thickness sample) was developed for the measurements of ginseng samples.
The analysis results of near infrared spectra also show that the epidermis, phloem and xylem parts of ginseng root are obvious different according to the NIR spectra, even spectra measured at different positions from top to bottom of epidermis, phloem and xylem show gradually changes.
Operation conditions of HPLC analysis for ginseng samples were established and the samples from the epidermis, phloem and xylem parts were analyzed. The classification results of the three parts due to HPLC data are similar with those of IR and NIR data. The results indicate that IR and NIR spectroscopy can replace HPLC to analyze the differences between different parts of epidermis, phloem and xylem of ginseng.
引文
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