HPLC测定非冷冻人全血中吲达帕胺及相对生物利用度

HPLC determination of indapamide and relative bioavailability in unfreezed human blood

目的:建立人全血中吲达帕胺的 HPLC—UV 测定法,研究2种吲达帕胺片的相对生物利用度。方法:全血样品液液萃取后,经 Diamonsil C_(18)柱(4.6 mm×200 mm,5 μm)分离,240 nm 检测,流动相为乙腈-0.1%三乙胺溶液(磷酸调 pH 至4.0)(38:62,v/v),流速为1.0mL·min^(-1)。18名健康志愿者采用随机交叉方式分别单次口服吲达帕胺片 T 或 R 5 mg,在不同时间点取血,样品以新建立的 HPLC—UV 法测定,研究比较2制剂的药动学及相对生物利用度。结果:吲达帕胺与全血中内源性杂质分离度好,吲达帕胺浓度住3.1～500μg·L^(-1)范围内与峰面积比线性良好,最低定量浓度为3.1μg·L^(-1)。方法回收率为95.6%～102.6%,日内精密度(RSD)小于14.2%,日间精密度(RSD)小于9.9%。2种制剂的 C_(max)为(250.9±84.1)μg·L^(-1)和(245.4±78.8)μg·L^(-1);T_(max)为(2.4±0.5)h 和(2.4±0.5)11;AUC_(0-72)为(4417.4±976.3)μg·h·L^(-1)和(4372.7±942.0)μg·h·L^(-1);T_(1/2)为(21.9±8.6)h 和(21.4±5.5)h。结论:该方法选择性好,灵敏度高,可用于吲达帕胺的体内过程研究。药动学参数经方差分析表明吲达帕胺片 T 和 R 中吲达帕胺的主要药动学参数之间均无明显差异,双单侧 t 检验结果表叫2种制剂为尘物等效制剂。

Objective：To develop an HPLC -UV method for the determination of indapamide in unfreezing blood and evaluate the bioequivalence of indapamide tablets. Methods： Sample preparation based on liquid - liquid extraction. The compounds were separated on a Diamonsil C,s column （4. 6 mm× 200 mm,5μm）with acetonitrile - 0. 1% triethylamine （ pH 4.0） （38： 62,v/v） as mobile phase at a flow rate of 1.0 mL ·min^- 1, and detected at 240 nm. In a randomized crossover design 18 healthy male volunteers were given 5 mg indapamide tablets T （test drug） or tablets R （reference drug）. The blood samples were obtained and determined by the newly - developed HPLC - UV method and the pharmacokinetics and bioavailability were studied. Results：The calibration curve of indapamide was linear over the range of 3.1 - 500 μg·L^-1 with the limit of quantity 3.1 μg·L^-1, the methodology recoveries were in the range of 95.6% - 102.6%. Inter - day and intra - day RSDs were less than 14. 2% and 9.9% ,respectively. Pharmacokinetic parameters of indapamide after a single dose of tablets T and R were as following ： Cmax were （250.9 ±84. 1 ）μg·L^-1 and （245.4 ±78.8） μg·L^-1,T were（ 2.4 ±0.5） h and（ 2.4 ±0.5） h,AUC0-72 were （4417.4±976.3）μg·h·L^-1 and （4372.7 ±942.0） μg·h· L^-1,T1/2 were （21.9 ±8.6） h and（ 21.4 ± 5.5 ） h. Conclusions ： The method is selective, sensitive and rapid for indapamide determination. The results of analysis of variance and two one - side t - test analysis show that no significant differences between the pharnlacokinetic parameters of the two formulations and bioequivalence are observed.