高效液相層析法檢測血清中9,10──蔥醌酮－2－羧酸濃度及其在藥物動力學研究之應用   全文下載

HPLC Determination of 9,10-Anthraquinone-2-Carboxylic Acid in Serum and Its Application to Pharmacokinetic Study

本研究發展出一簡單、高感度和精確之高效液相層析法，用以檢測家兔血清中9,10－蔥?酮－2－羧酸（AQCA）濃度。本分析法以LichrosherR 100RP-18為層析管柱，0.4%磷酸：乙氰（7:3） ／ 甲醇 = 45:55 為移動相，流速為1.2 mL ／ min，紫外光檢測器之檢測波長為256 nm，甲基苯甲酸為內標準品。檢品經簡單處理後，注入管柱分析。最低檢測值為0.6 μg／mL，於0.6～18μg／mL 濃度間具有良好的線性檢量關係，確效試驗顯示，精密度佳（變異係數少於2%），回收率為96.98%。繼以此分析法應用於藥品動力分析研究之探討。十八隻家兔分成三組，取不同劑型之AQCA，以相同的單一劑量（10 mg／kg），分別以血管注射、口服和直腸投藥，檢測各組各時段血清中的AQCA濃度，經Winnolin軟體處理資料，並以ANOVA試驗（α = 0.05）評定各組重要的藥品動力參數之異同。注射投藥時，半衰期（t1／2）、血清濃度時間曲線下面積（AUC）分別為4.43 ± 0.13 hrs和141.79 ± 7.84μg* hr／mL。口服投藥的參數數據為血清中尖峰濃度（Tmax） = 7.17 ± 0.41 hrs； t1／2 = 15.32 ± 1.86 hrs； 平均滯留時間（MRT） = 20.02 ± 2.07 hrs； AUC = 107.53 ± 4.50μg* hr／m，顯示口服途徑的藥品吸收速率慢、滯留時間長、吸收總量低。直腸投藥的參數數據為Tmax = 1.58 ± 0.20 hrs； t1／2 = 5.67 ± 0.82 hrs； AUC = 121.18 ± 6.19μg* hr／mL，依數據比較，藥物吸收速率及吸收總量均比前者好。另者，在本劑量下，口服和直腸投藥的絕對生體可用率分別為0.876 和0.872。這些參數上的差異可能是AQCA於不同的劑型與投藥部位、有不同的溶離性質所致。"

A simple and sensitive high-performance liquid chromatographic (HPLC) method involving UV detection was developed for the determination of 9,10-anthraquinone-2-carboxylic acid (AQCA) in rabbit serum. A reverse-phase column (Lichrosher R 100RP-18) was eluted with a mobile phase of 0.4% phosphoric acid: acetonitrile (7:3) / methanol = 45 : 55 at a flow rate of 1.2 mL/min. The UV absorbance was monitored at 256 nm. After a simple clean-up procedure, the limit of quantitation achieved was 0.6 μg/mL and the standard curve was found to be linear over the serum concentration of 0.6 ~ 18μg/mL. The intra-assay and inter-assay coefficient of variance in serum was less than 2%, and the recovery was 96.98%. The established HPLC method was applied to the study of pharmacokinetics and bioavailability of 9,10-anthraquinone-2-carboxylic acid (AQCA). Eighteen healthy rabbits were divided into three groups and given intravenous (i.v.) , oral and rectal administrations of different AQCA preparations with a single dose of 10 mg/kg. Blood samples were collected and AQCA concentrations in serum were analyzed. The results were performed with Winnolin programs and the ANOVA test (α = 0.05) was used to compare the pharmacokinetic parameters of AQCA in the three regimens. For the i.v. route, the t1/2 and AUC were 4.43 ± 0.13 hrs and 141.79 ± 7.84 μg*hr/mL, respectively. Parameters for the oral route were : Tmax = 7.17 ± 0.41 hrs ; t1/2 = 15.32 ± 1.86 hrs; MRT = 20.02 ± 2.07 hrs and AUC = 107.53 ± 4.50 μg*hr/mL. The oral route indicated a slower absorption rate, a longer residence time and a lower extent of the AQCA absorption. For the rectal route, Tmax occurred at 1.58 ± 0.20 hrs, the t1/2 was 5.67 ± 0.82 hrs and the AUC was 121.18 ± 6.19 μg*hr/mL. The results indicated that the rate and extent of AQCA rectal absorption were better compared to those of the oral route. At this dose, the absolute bioavailabilities of AQCA were 0.876 and 0.872 for oral and rectal administrations, respectively. The differences of these pharmacokinetic parameters might be due to the physicochemical properties of AQCA in different conditions. 本研究發展出一簡單、高感度和精確之高效液相層析法，用以檢測家兔血清中9,10－蔥醌酮－2－羧酸（AQCA）濃度。本分析法以LichrosherR 100RP-18為層析管柱，0.4%磷酸：乙氰（7:3） ／ 甲醇 = 45:55 為移動相，流速為1.2 mL ／ min，紫外光檢測器之檢測波長為256 nm，甲基苯甲酸為內標準品。檢品經簡單處理後，注入管柱分析。最低檢測值為0.6 μg／mL，於0.6～18μg／mL 濃度間具有良好的線性檢量關係，確效試驗顯示，精密度佳（變異係數少於2%），回收率為96.98%。繼以此分析法應用於藥品動力分析研究之探討。十八隻家兔分成三組，取不同劑型之AQCA，以相同的單一劑量（10 mg／kg），分別以血管注射、口服和直腸投藥，檢測各組各時段血清中的AQCA濃度，經Winnolin軟體處理資料，並以ANOVA試驗（α = 0.05）評定各組重要的藥品動力參數之異同。注射投藥時，半衰期（t1／2）、血清濃度時間曲線下面積（AUC）分別為4.43 ± 0.13 hrs和141.79 ± 7.84μg* hr／mL。口服投藥的參數數據為血清中尖峰濃度（Tmax） = 7.17 ± 0.41 hrs； t1／2 = 15.32 ± 1.86 hrs； 平均滯留時間（MRT） = 20.02 ± 2.07 hrs； AUC = 107.53 ± 4.50μg* hr／m，顯示口服途徑的藥品吸收速率慢、滯留時間長、吸收總量低。直腸投藥的參數數據為Tmax = 1.58 ± 0.20 hrs； t1／2 = 5.67 ± 0.82 hrs； AUC = 121.18 ± 6.19μg* hr／mL，依數據比較，藥物吸收速率及吸收總量均比前者好。另者，在本劑量下，口服和直腸投藥的絕對生體可用率分別為0.876 和0.872。這些參數上的差異可能是AQCA於不同的劑型與投藥部位、有不同的溶離性質所致。