Study on the High Performance Liquid Chromatography Applied to Pharmaceutical Analysis

Abstract: High performance liquid chromatography (HPLC) as an important modern measurement of separation now is one of high performance and fastly speed analysis and separation technology emerging in the 1970's. Due to its' convenience, high-speed, sensitivity and fairly accuracy, it has been widely used in the areas of production of medicine, foodstuff, and environment etc.This thesis establishes several detection methods by Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC). It includes two sections—review and research report. In the review section, information about high performance liquid chromatography such as history, principles, HPLC equipment and the applications in the medicine analysis was summarized. In the next research report section, analysis methods of several compounds in the taita beauty essence and beeswax tablets were reported. That is: Respective determination of ferulic acid, cinnamic acid and quercetin in taita beauty essence by reversed-phase high-performance liquid chromatography; Determination of caffeic acid in beeswax tablets and simultaneous determinations of five compositions in beeswax tablets by reversed-phase high-performance liquid chromatography.1. Determination of Ferulic Acid in Taita Beauty Essence by Reversed-Phase High-Performance Liquid ChromatographyThe HPLC method for the determination of ferulic acid in Taita Beauty Essence was established. A Shim-pack VP-ODS column ( 150 x 4.6 mm, 5 urn ) was used, with the mobile phase of methanol-water-acetic acid (32: 66: 2) at the detection wavelength of 323 nm. The flow rate was 1 ml/min. The calibration curve was linear in the range of 0.252~5.04μg/ml (r=0.9993 ). The average recovery was 99.9 % ( n=5 ) and RSD was 1.2 %. It was proved that the method was convenient, rapid, accurate and suitable for the determination of ferulic acid in Taita Beauty Essence. 2 Determination of Cinnamic Acid in Taita Beauty Essence by Reversed-Phase High-Performance Liquid ChromatographyA method using HPLC was developed for determination of cinnamic acid in taita beauty essence. The separation was carried on a Shim-pack VP-ODS column (150×4.6 mm, 5μm ) and a guard column (10×4.6 mm, 5.0μm), with the mobile phase of methanol-water-acetic acid=( 70: 29: 1,V/V ) at the detection wavelength of 280 nm. The flow rate was 1 ml/min. The calibration curve was linear in the range of 0.012~4.80μg/mL, (r=0.9998 ). The average recovery was 99.1 % (n=5 ) and RSD was 3.26 %. The method has been successfully applied to determination, which was proved to be simple and rapid.3 Determination of Quercetin in Taita Beauty Essence by Reversed-Phase High-Performance Liquid ChromatographyIn this chapter, a HPLC method was developed for the determination of quercetin in Taita Beauty Essence. A Shim-pack VP-ODS column (150×4.6 mm, 5μm ) and a guard column (10×4.6 mm, 5.0μm) were used, The analysis was carried under the condition of mobile phase of methanol-0.2 % (v/v) phosphoric acid in water ( 60:40) and at the detection wavelength of 254 nm. The flow rate was 1 ml/min. The calibration curve was linear in the range of 0.00720~0.360μg/mL (r=0.9999 ). The average recovery was 93.9 %~98.6 % (n=5) and the relative standard deviation was 2.07 %. The method was suitable for the determination of quercetin in Taita Beauty Essence. A good result was obtained.4. Determination of Caffeic Acid in Beeswax Tablets by Reversed-Phase High-Performance Liquid ChromatographyThe HPLC method for the determination of caffeic acid in beeswax tablets was established. A Shim-pack VP-ODS column (150×4.6 mm, 5μm) and a guard column (10×4.6 mm, 5.0μm) were used, with the mobile phase of methanol-water-formic acid (35: 64: 1) at the detection wavelength of 323 nm. The flow rate was 1 ml/min. The calibration curve was linear in the range of 0.120~24.0μg/mL (r=0.9995 ). The average recovery was 101.0 % (n=5) and the relative standard deviation was 2.69 %. It was proved that the method was convenient, rapid and suitable for the determination of caffeic acid in Taita Beauty Essence.5. Simultaneous Determinations of Five Components in Beeswax Tablets by Reversed-Phase High-Performance Liquid ChromatographyA reversed high-performance liquid chromatographic method was employed to the determination of ferulic acid, benzoic acid, salicylic acid, cinnamic acid and quercetin in beeswax tablets. These five compounds were analyzed simultaneously with a shim-pack VP-ODS (150×4.6 mm, 5.0μm) and a guard column (10×4.6 mm, 5.0μm) by gradient elution using 0.2 % (v/v) phosphoric acid in water-methanol as the mobile phase. The flow rate was 1 ml/min and the detection wavelength was performed according to time program: 0-20 min, 323 nm; 20-25 min, 230 run; 25-37 min, 236 nm; 37-45 min, 280 nm; 45-60 min, 254 nm. The recovery of the method was in the range of 94.2 %-111.2 %, and all the compounds showed good linearity (r-0.9979-0.9996) in a relatively wide concentration range. The method has been successfully applied to determination of the five compounds in beeswax tablets…
Key words: high-performance liquid chromatography; medicine analysis; beeswax tablet; taita beauty essence

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