Simultaneous quantification of seven bioactive components in Caulis Lonicerae Japonicae by high performance liquid chromatography

This study presents a new HPLC method for the simultaneous determination of seven major components, namely chlorogenic acid, caffeic acid, loganin, sweroside, secoxyloganin, rutin and luteolin 7-O-glucoside in Caulis Lonicerae Japonicae, a commonly used traditional Chinese medicinal herb derived from the caulis of Lonicera japonica Thunb. These seven compounds, belonging to the chemical types of phenolic acids, iridoids and flavonoids, were separated on a C18 column (250 x 4.6 mm, 5.0 microm) with the column temperature at 30 degrees C. The mobile phase was composed of (A) aqueous acetic acid (0.4%, v/v) and (B) acetonitrile using a gradient elution of 10% B at 0-12 min, 10-17% B at 12-25 min and 17% B at 25-35 min. The flow rate was 1.0 mL/min and detection wavelength was set at 245 nm. The limit of detection (S/N = 3) ranged from 0.10 to 0.23 microg/mL and the limit of quantification (S/N = 10) ranged from 0.69 to 3.56 microg/mL. All calibration curves showed good linear regression (r2 > 0.9990) within the test ranges. The intra- and inter-day precisions as determined from sample solutions were below 1.24 and 2.28%, respectively. The recoveries for seven compounds were found to range from 94.2 to 103.6%. This verified method has been successfully applied to evaluation of commercial samples of Caulis Lonicerae Japonicae from different markets in China.     

Binding study of Flos Lonicerae Japonicae with bovine serum albumin using centrifugal ultrafiltration and liquid chromatography

The screening and analysis of bioactive components in traditional Chinese medicines (TCMs) is very important not only for the quality control of Chinese herbs but also for elucidating the therapeutic principles. This study developed a new method for screening and analyzing bioactive compounds from TCMs using centrifugal ultrafiltration coupled with high-performance liquid chromatography. The method was successfully applied in the binding study of Flos Lonicerae Japonicae with bovine serum albumin (BSA), and 11 compounds were found to be bound with the BSA. Eight of them were positively identified as chlorogenic acid, caffeic acid, rutin, quercetin-3-O-glucoside, luteolin-7-O-glucoside, lonicerin, 3, 5-di-O-caffeoyl quinic acid and 3,4-di-O-caffeoyl quinic acid. Another three compounds were tentatively identified as two isomers of chlorogenic acid and one isomer of di-O-caffeoyl quinic acid by comparing the UV data and MS data with the previous reports. Based on modern pharmacological study, these compounds are the major bioactive components in Lonicera japonica. Therefore, the proposed method could be a good approach to predicting the potential bioactivities of multiple compounds in TCMs simultaneously.     

Development of microwave-assisted extraction followed by headspace solid-phase microextraction and gas chromatography-mass spectrometry for quantification of camphor and borneol in Flos Chrysanthemi Indici

In the work, microwave-assisted extraction (MAE) followed by headspace solid-phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC-MS) was developed for quantitative analysis of the bioactive components of camphor and borneol in a traditional Chinese medicines (TCM) of Flos Chrysanthemi Indici. After systematical investigation, the optimal experimental parameters microwave power (400 W), irradiation time (4 min), fiber coating (PDMS/DVB fiber), extraction temperature (40 °C), extraction time (20 min), stirring rate (1100 rpm), and salt effect (no salt added) were investigated. The optimized method provided satisfactory precision (RSD values less than 12%), good recovery (from 86% to 94%), and good linearity (R² > 0.999). The proposed method was applied to quantitative analysis of camphor and borneol in Flos Chrysanthemi Indici samples from 11 different growing areas. To demonstrate the method feasibility, steam distillation was also used to analyze camphor and borneol in Flos Chrysanthemi Indici samples from these different growing areas. The very close results were obtained by the two methods. It has been shown that the proposed ME-HS-SPME-GC-MS is a simple, rapid, solvent-free and reliable method for quantitative analysis of camphor and borneol in TCM, and a potential tool for quality assessment of Flos Chrysanthemi Indici.     

高效液相色谱法测定野菊花中蒙花苷的含量

建立高效液相色谱法测定野菊花中蒙花苷含量的方法。采用KromosilTM C18十八烷基硅烷键合硅胶色谱柱,甲醇-水-乙酸(体积比为55∶44.5∶0.5)为流动相,流速为0.8mL/min,柱温为室温,检测波长为334nm。蒙花苷含量在11.2~89.6μg/mL范围内,线性关系良好(r=0.9998),方法的相对标准偏差为0.51%(n=5),平均加标回收率为98.92%。