ABSTRACT
The present study determined the quantitative markers of total proanthocyanidins in the purification of the industrial waste Choerospondias axillaris pericarp based on the comparison results of high-performance liquid chromatography(HPLC) and mass spectrometry(MS) and optimized the purification process with two stable procyanidins as markers. The adsorption and desorption of five different macroporous adsorption resins, the static adsorption kinetics curve of NKA-Ⅱ resin, the maximum sample load, and the gradient elution were investigated. The UPLC-Q-TOF-MS/MS was employed for qualitative analysis of the newly-prepared total proanthocyanidins of C. axillaris pericarp. As revealed by the results, NKA-Ⅱ resin displayed strong adsorption and desorption toward total proanthocyanidins. The sample solution(50 mg·mL~(-1)) was prepared from 70% ethanol crude extract of C. axillaris pericarp dissolved in water and 7-fold BV of the sample solution was loaded, followed by static adsorption for 12 h. After 8-fold BV of distilled water and 6-fold BV of 10% ethanol were employed to remove impurities, the solution was eluted with 8-fold BV of 50% ethanol, concentrated, and dried under reduced pressure, and purified total proanthocyanidin powder was therefore obtained. Measured by vanillin-hydrochloric acid method, the purity and transfer rate of total proanthocyanidins were 47.67% and 59.92%, respectively, indicating the feasibi-lity of the optimized process. UPLC-Q-TOF-MS/MS qualitative analysis identified 16 procyanidins in C. axillaris total proanthocyanidins. The optimized purification process is simple in operation and accurate in component identification, and it can be applied to the process investigation of a class of components that are difficult to be separated and purified. It can also provide technical support and research ideas for the comprehensive utilization of industrial waste.
Subject(s)
Adsorption , Anacardiaceae , Chromatography, High Pressure Liquid , Plant Extracts , Proanthocyanidins/analysis , Resins, Synthetic , Tandem Mass SpectrometryABSTRACT
OBJECTIVE：To establish a method for the content determination of to tal flavonoids from Amomum tsao-ko ，and to optimize the purification technology by macroporous resin. METHODS ：The content of total flavonoids was measured by HPLC. The determination was performed on Eclipse Plus C 18 column with mobile phase consisted of acetonitrile- 1％ acetic acid solution （15∶85，V/V）at the flow rate of 0.8 mL/min. The column temperature was 40 ℃，and the detection wavelength was set at 256 nm. The sample size was 10 μL. Taking the adsorption and desorption performance as indexes，6 kinds of macroporous resins were screened out by static adsorption and desorption tests ；adsorption and desorption time were investigated by static adsorption and desorption kinetics tests. Using the content of total flavonoids （calculated by rutin ）as index ，with sample concentration ，sample pH，ethanol volume fraction and elution amount as factors ，based on single factor test ，orthogonal design was used to optimize the purification technology of total flavonoids from A. tsao-ko ，and validation test was performed. RESULTS ：The linear range of rutin were 0.028-0.281 mg/mL（r＝0.999 9）. The limit of quantification was 437.5 ng/mL and the limit of detection was 109.4 ng/mL. RSDs of precision ，stability and reproducibility tests were all lower than 2％；the recoveries were 96.24％-99.75％（RSD＜2％，n＝ 6）. The comprehensive capacity of adsorption and desorption of HPD 450 macroporous resin was the most suitable ，and the best static adsorption and desorption time both were 12 h. The optimal purification technology was 1.854 4 mg/mL ； ethanol elution was 8 times of the column volume. Vertificationtests show that after optimized ，the content of total flavonoids from A. tsao-ko increased from 22.556 7 mg/g to 57.728 2 mg/g. The purity of was 2.56 and stable for the content determination. Optimal purification technology is stable and feasible ，which is suitable for purifieation of total flavonoids from A. tsao-ko .
ABSTRACT
Objective To optimize separation and purification technology of Zhizichi decoction by macroporous resina . Methods The content of total iridoid glycosides and total isoflavones were determined by ultraviolet spectrophotometry .Tak-ing the content of total iridoid glycosides and total isoflavones as indexes ,the effect of the concentration of sample solution , medicinal herbs on the amount of sample ,concentration and volume of eluent was investigated by single factor test .Results The optimum separation and purification technology was as following :the concentration of sample solution was 0 .1 g/ml ,the volume ratio of resin to material drug was 2∶1 ,the adsorption time was 2 h ,and the sample was firstly eluted with 1 BV wa-ter ,then 6 BV 20% ethanol and 60% ethanol ,and the eluent was collected .Conclusion This optimized separation and purifi-cation technology was reasonable and stable ,and it could be extended to large-scale production applications .
ABSTRACT
Objective To study the chemical constituents in the immature fruits of Momordica charantia. Methods Isolation and purification were carried out by macroporous absorption resin and silica gel, and compounds were identified and elucidated by spectral and chemical methods. Results Ten compounds were obtained from alcohol extract and five of them were determined as germanicyl acetate (Ⅰ), aglycone of momordicoside Ⅰ (Ⅱ), aglycone of momordicoside L (Ⅲ), charantin (Ⅳ), and ?-sitosterol (Ⅴ). Conclusion For all the first time, compound Ⅰ is found in the plants of Momordica L., compound Ⅱ is found as novel natural product in this plant, and compound Ⅲ is found in China.
ABSTRACT
AIM: To study the purification of Naomaitong Granules(Radix et Rhizoma rhei,Rhizoma chuanxiong,Radix Puerariae lobatae.etc) by macroporous absorption resin. METHODS: Naomaitong Granule was purificated by macroporous absorption resin AB-8.UV spectrothotometry was used to determine the contents of total anthraquinones,total ginsenosides,total alkaloids,and the content of puerarin was determined by HPLC.The technic of purification was optimized according to the content above. RESULTS: The optimized technological conditions consisted of eoncentranon of original sample 120 mg/mL,the diameter and height was in proportion of 1∶10,the ratio of maximum adsorption to resin volume was 1∶6,water elution was 2B multiple of resin volume,8B multiple of resin volume 50% alcohol was the elution. CONCLUSION: AB-8 macroporous absorption resin can be used to purify Naomaitong Granules.