| The dried flower of safflower (Carthamus tinctorius L), a common huoxuehuayu remedy in the traditional Chinese medicine, has been used for gyaechological diseases and heart diseases and sedative and anti-inflammatory. A major ingredient in the flower of safflower, which has been found to possess pharmacological effects, is safflor yellow. Safflor yellow A is the main component of safflor yellow. Until now, the report of safflor yellow A has focused on pharmacology and extraction, but there are few studies about its analysis and pharmacokinetics. In this study, the methods for the extraction and separation of safflor yellow A from the flower of safflower, and the methods for the determination of safflor yellow A in the flower of safflower and safflowers injection were established, and the pharmacokinetics of safflor yellow A in rats and the distribution of safflor yellow A in tissue of mouse were also studied.1. The safflor yellow A was separated from the aqueous extraction of safflower by column chromatography on polyamide and macroporous absorption resin, followed by preparative liquid chromatography.2. Two high performance liquid chromatographic methods (HPLC) for the determination of safflor yellow A in the flower of safflower and safflowersinjection were established, respectively. In the method for the determination of safflor yellow A in the flower of safflower, the Hypersil ODS2(200 mmx 4.6 mm,5um) column and Methanol-0.2 % acetic acid (10:90,v/v) as mobile phase were used, and Vanillic acid was used as the internal standard. The flow rate was 1.0 mL·min-1. The UV detection wavelength was set at 222 nm. The calibration curve was linear in the range from 9.4 ug·mL-1 to 150.4 Hg·mL-1, with r = 0.9998. The limit of detection was 0.087 ug·mL-1. The recovery was 97.6 %. The relative standard deviation (RSD) was 1.7 %. In the method for the determination of safflor yellow A in safflowers injection, the analytical column was Hypersil ODS2 column (200 mm×4.6mm, 5um). The mobile phase consisted of Methanol-0.2% acetic acid (13:87,v/v) at the flow rate of 1.0 mL·min-1. The UV detection wavelength was 402 nm. The calibration curve was linear in the range from 4.0ug·mL-1 to 80.8 ug-mL-1, with r =0.9996. The limit of detection was 0.055 ug·mL-1. The recovery was 97.7% , and RSD was 1.6%. And the content of safflor yellow A in the flower of safflower and safflowers injection were determined, respectively, by using those methods.3. A method for the determination of the concentration and pharmacokinetic parameters of safflor yellow A in plasma of rats were established by using HPLC. The analytical column was Hypersil ODS2 column (200 mm× 4.6 mm, 5um). The mobile phase consisted of Methanol-0.2% acetic acid (22:78,v/v). The flow rate was 0.8 mL·min-1. The UV detection wavelength was set at 402 nm. Riboflavin was used as internal standard. The calibration curve was linear in the range from 0.45 ng·mL-1 to35.60 ug·mL-1, with r = 0.9993, and the limit of detection was 0.060ug·mL-1.The RSDs of within-day and between-day were both below 10.0 %. Aftersingle dose (6.72 mg·kg-1) of safflor yellow A was injected into the rats through caudal vein, the primary pharmacokinetic parameters were estimated with the 3P87 software. The primary pharmacokinetic parameters were as follows: Vc :0.30±0.04 L·kg-1, CL : 1.12 + 0.33 mL·min-1, Ke : 0.91±0.19 h-1, t1/2 : 0.76±0.10 h, AUC :24.97±4.83 ug·h·mL-1, respectively. The metabolism of safflor yellow A in rats fit to one-compartment model. It distributed and eliminated quickly.4. An HPLC method for the determination of safflor yellow A in tissue of mouse was developed. Hypersil ODS2(200rnm × 4.6rnm,5um) column and Shimadzu YWG-ODS(10 mm ×4.6mm) pre-column were used. The mobile phase consisted of Methanol-0.2% acetic acid (24:76, v/v) at the flow rate of 0.8 mL·min-1. The UV detection wavelength was set at 402 nm. Pyrazolone was used as internal standard. The RSDs of within-day and between-day were all less than 8.0%. The characteri... |
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