Determination Of The Content Of The Diarrhea Type Traditional Chinese Medicine Preparations By SPE-HPLC

The determination of Chinese medicine effective composition content is the core part of the quality control system, this study establish a new approach for the determination of the content of active ingredients in 5 kinds of diarrhea type Chinese medicine compound preparations by using high performance liquid chromatography(HPLC) method, including Changweining Capsule, Xielixiao Tablet, Xieliguchang Tablet, Er’bao Particle and Bupiyichang Pill. Meanwhile combine with solid phase extraction(SPE) pretreatment technology separate and purify the samples, to separate the impuritiy and the target effectually, and improve the recovery rate. The method is simple, less impurity interference, high sensitivity, accurate and reliable, can provide effective basis for quality control of diarrhea type Chinese medicine compound preparations. This thesis mainly has the following six chapters content:The first chapter ReviewsThrough literature research, summarize the research status about the diarrhea class content of effective components in traditional Chinese medicine compound preparations in recent years, pretreatment technology, determination method and mechanism, simultaneously, discusses the development prospects of the high performance liquid chromatography and solid phase extraction.The second chapter Determination of 4 kinds of effective components in Xielixiao Tablets by SPE-HPLCTo establish a solid-phase extraction with high-performance liquid chromatography(SPE-HPLC) method for determination of paeoniflorin, liquiritin, naringin and hesperidin in Xielixiao Tablet. Through a series of optimization of chromatographic conditions and solid phase extraction conditions. Using C18 solid phase extraction column to purge and enrichment the active ingredients of Xielixiao Tablet. The contents was analyzed by HPLC method with Kromasil C18 column(250mm×4.6mm, 5μm). The mobile phase composed of acetonitrile and 5m L·L-1 phosphoric acid in gradient elution. Detection wavelength were set at 230 nm, 276 nm and 284 nm. The column temperature was maintained at 28℃, and injection volume was 20μL. The flow rate was 1.0m L·min-1. The calibration curves of paeoniflorin, liquiritin, naringin, hesperidin respectively showed good linearity within 0.6170~30.85μg·m L-1(r=1.0000), 0.1598 ~ 7.944μg·m L-1(r=0.9998), 0.9728 ~ 48.64μg·m L-1(r=1.0000), 0.1236 ~6.180μg·m L-1(r=0.9999). The average recoveries of these were 99.4%, 98.3%, 97.8% and 97.7%,RSDs were 0.97%, 1.11%, 1.28% and 1.26%, respectively.The third chapter Simultaneous Determination of 6 kinds of effective components in Changweining Capsule by HPLCThrough the optimization of chromatographic conditions, confirm the best extraction solvent, solid-liquid ratio and ultrasonic time. To investigate the identification of six kinds effective components in Changweining Capsule by HPLC, these components are puerarin, peoniflorin, liquiritin, psoralen, isopsoralen and costunolide. The HPLC column is the Kromasil C18 column(4.6mm×250mm, 5μm). The mobile phase composed of acetonitrile and 0.5% phosphoric acid in gradient elution. Detection wavelength were set at 230, 276, 246 and 225 nm. The flow rate was 1.0m L·min-1. The injection volume was 20μL. The calibration curves of puerarin, paeoniflorin, liquiritin, psoralen, isopsoralen and costunolide respectively showed good linearity within 1.650 ~82.51μg·m L-1, 0.4628~23.14μg·m L-1, 0.3178~15.89μg·m L-1, 0.5818~29.09μg·m L-1, 0.4864~24.32μg·m L-1, 0.3280~16.40μg·m L-1. The average recoveries of these were 96.9%, 97.2%, 97.5%, 96.0%, 95.2%, 97.2%, respectively.The fourth chapter Determination of Paeoniflorin, Liquiritin and Hesperidin in Xieliguchang Tablet by SPE-HPLCTo establish a method for determination of three kinds effective components in Xieliguchang Tablet with Kromasil C18 column separation and Hyper Sep C18 solid phase extraction column purification the samples by SPE-HPLC. The mobile phase composed of acetonitrile and 0.1% phosphoric acid(24:76). Detection wavelength were set at 230nm(paeoniflorin, liquiritin) and 284nm(hesperidin). The column temperature was maintained at 25℃, and injection volume was 20μL, the flow rate was 1.0m L·min-1. The calibration curves of paeoniflorin, liquiritin and hesperidin respectively showed good linearity within 3.998~199.8μg·m L-1, 0.6356~31.78μg·m L-1, 0.4940~24.70μg·m L-1. The repeatability and stability RSDs were less than 3%. The average recoveries of these were 98.6%, 97.0%, 98.0%, respectively.The fifth chapter Determination of Puerarin, Paeoniflorin and Hesperidinthe in Er’bao Particle by HPLCTo investigate a method for the identification of puerarin, paeoniflorin and hesperidinthe three kinds effective components in Er’bao Particle by HPLC, in order to provide a scientific basis to control its quality standard. Using Kromasil C18 column(4.6mm×250mm, 5μm). The mobile phase composed of acetonitrile and 0.5% phosphoric acid in gradient elution(0~5min, 24% A; 5~10min, 24% Aâ†'28% A). Detection wavelength switching were 0~4.0min, 250nm; 4.0~6.0min, 230nm; 6.0~10min, 284 nm. The flow rate was 1.0m L·min-1. The column temperature was maintained at 28℃. The injection volume was 20μL. Results, the calibration curves of Puerarin, paeoniflorin and hesperidinthe respectively showed good linearity within 3.098~77.44μg·m L-1, 3.456~86.40μg·m L-1, 0.4416~11.04μg·m L-1. The average recoveries of these were 99.1%, 98.6%, 98.2%, respectively. RSDs were 1.76%, 1.02%, 1.23%, respectively.The sixth chapter Determination of psoralen, isopsoralen and peoniflorin in Bupiyichang Pill by SPE-HPLCTo establish a method for determination of three kinds effective components in Bupiyichang Pill, with Hyper Sep Retain PEP solid phase extraction small column to purge and enrichment the psoralen, isopsoralen and peoniflorin of Bupiyichang Pill. The contents was analyzed by HPLC method with Kromasil C18 column(4.6mm×250mm, 5μm). The mobile phase composed of acetonitrile and 0.1% phosphoric acid in gradient elution. The flow rate was 1.0m L·min-1. Detection wavelength were set at 246 nm and 230 nm. The column temperature was maintained at 25℃, and injection volume was 20μL. The calibration curves of psoralen, isopsoralen and peoniflorin respectively showed good linearity within 0.8726 ~ 43.63μg·m L-1(r=0.9999), 1.216 ~60.80μg·m L-1(r=0.9998), 2.912~145.6μg·m L-1(r=0.9997). The average recoveries of these were 97.6%, 97.2%, 97.5%, respectively. RSDs were 1.00%, 1.09%, 0.52%, respectively.