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基于高效液相色谱-蒸发光散射检测器特征图谱的地黄炮制前后成分变化研究

[Ingredients changes in Rehmanniae Radix processing based by HPLC-ELSD specific chromatogram].

作者信息

Zhang Wen-Ting, He Jia, Guo Zeng-Xi, Shi Shang-Mei

机构信息

Zhenjiang Institute for Food and Drug Control Hangzhou 310052, China NMPA Key Laboratory for Quality Evaluation of Traditional Chinese Medicine (Traditional Chinese Patent Medicine) Hangzhou 310052, China.

College of Pharmaceutical Sciences, Zhejiang Chinese Medical University Hangzhou 310053,China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2020 Aug;45(16):3877-3882. doi: 10.19540/j.cnki.cjcmm.20200523.303.

Abstract

To establish the HPLC-ELSD specific chromatogram analysis method of Rehmanniae Radix and Rehmanniae Radix Prae-parata, and analyze and compare their chemical compositions, so as to reveal the change regularity of compositions during the proces-sing. By HPLC-ELSD method, the chromatographic column for Prevail Carbohydrate ES(4.6 mm ×250 mm, 5 μm) was adopted, with acetonitrile(A)-water(B) as mobile phase for gradient elution, and the evaporative light-scattering detector was used. A total of 23 batches of Rehmannia Radix samples, and 25 batches of Rehmanniae Radix Praeparata samples and processing dynamic samples were compared. The established method had a great repeatability, precision and stability. Eight common chromatographic peaks were extracted from 23 batches of Rehmanniae Radix samples, 8 common peaks were extracted from 25 Rehmanniae Radix Praeparata, and 7 chromatographic peaks were identified. The composition ratio of Rehmannia Radix was changed greatly during the processing. When the simila-rity≥0.95 and the fructose peak area was more than 2 times of stachyose tetrahydrate or more than 20 times of raffinose, the processing degree conformed to the requirements of empirical identification. The three main oligosaccharides of Rehmanniae Radix were sucrose that was heated to generate fructose and glucose, stachyose tetrahydrate that was heated to generate melibiose, sucrose and fructose, and stachyose tetrahydrate that was heated to generate manninotriose. The change in the index of proportion between monosaccharides and oligosaccharides can be used as the quantitative criterion for the processing quality of Rehmanniae Radix Praeparata.

摘要

建立生地黄和熟地黄的高效液相色谱-蒸发光散射检测(HPLC-ELSD)特征图谱分析方法,分析比较二者的化学成分,以揭示炮制过程中成分的变化规律。采用HPLC-ELSD法,色谱柱为Prevail Carbohydrate ES(4.6 mm×250 mm,5μm),以乙腈(A)-水(B)为流动相进行梯度洗脱,采用蒸发光散射检测器。比较了23批生地黄样品、25批熟地黄样品及炮制动态样品。所建立的方法具有良好的重复性、精密度和稳定性。从23批生地黄样品中提取出8个共有色谱峰,从25批熟地黄中提取出8个共有峰,并鉴定出7个色谱峰。生地黄在炮制过程中成分比例变化较大。当相似度≥0.95且果糖峰面积为水苏糖的2倍以上或棉子糖的20倍以上时,炮制程度符合经验鉴别要求。生地黄的三种主要低聚糖为加热生成果糖和葡萄糖的蔗糖、加热生成蜜二糖、蔗糖和果糖的水苏糖以及加热生成甘露三糖的水苏糖。单糖与低聚糖比例指标的变化可作为熟地黄炮制质量的定量标准。

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