秦向輝 常海民 甘莉 王彥 閆超
摘 要 目的:建立高效液相色譜方法測(cè)定前列地爾注射液中的有關(guān)物質(zhì)。方法:以辛烷基硅烷鍵合硅膠為填充劑的色譜柱(4.0 mm×250 mm,4 mm)、0.025 mol/L磷酸二氫鈉溶液(pH 2.5)為流動(dòng)相A、乙腈為流動(dòng)相B,梯度洗脫,流速1 mL/min,檢測(cè)波長(zhǎng)200 nm,柱溫35 ℃。結(jié)果:前列地爾在0.2~1.6 mg/mL的濃度范圍內(nèi)線性關(guān)系良好,檢測(cè)限為0.06 mg/mL,定量限為0.2 mg/mL,回收率為84.19%~103.30%,RSD為8.4%(n=9)。結(jié)論:前列地爾主成分色譜峰可與有關(guān)雜質(zhì)完全分離,方法專屬性強(qiáng),重復(fù)性好,可用于前列地爾注射劑有關(guān)物質(zhì)的檢查。
關(guān)鍵詞 前列地爾 有關(guān)物質(zhì) 注射劑 高效液相色譜
中圖分類號(hào):R972.4; R917 文獻(xiàn)標(biāo)志碼:A 文章編號(hào):1006-1533(2021)19-0074-05
Study on the determination method for the related substances in alprostadil injections
QIN Xianghui1, CHANG Haimin2, GAN Li3, WANG Yan1, YAN Chao1[ 1. School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China; 2. F.B.C. (Shanghai) Pharmaceutical Technology Co., LTD., Shanghai 201210, China; 3. School of Chemical & Environmental Engineering, Shanghai Institute of Technology, Shanghai 201418, China]
ABSTRACT Objective: To establish an HPLC method for the determination of related substances in alprostadil injections. Methods: HPLC was run on a Merck Superspher 60 RP-Select B column (4.0 mm×250 mm, 4 μm) at the flow rate of 1 mL/min, the detection wave length of 200 nm and column temperature 35 ℃ by gradient elution using 0.025 mol/L sodium dihydrogen phosphate solution (pH 2.5) and acetonitrile as mobile phase A and B. Results: The alprostadil showed good linearity over the range of 0.2-1.6 mg/mL with detection limit 0.06 mg/mL and quantitation limit 0.2 mg/mL, recovery 84.19%-103.30% and RSD (n=9) 8.4%. Conclusion: The chromatographic peaks of main components in alprostadil can be completely separated from the related impurities. The established method is highly specific and reproducible and can be used for the inspection of related substances in alprostadil injection.
KEy wORDS alprostadil; related substances; injections; HPLC
前列地爾是廣泛存在于體內(nèi)的生物活性物質(zhì)[1],前列地爾注射液是1988年在日本上市的前列地爾脂肪乳制劑[2],臨床上主要用于治療慢性動(dòng)脈閉塞癥、血栓閉塞性脈管炎、閉塞性動(dòng)脈硬化癥等引起的四肢潰瘍及微小血管循環(huán)障礙引起的四肢靜息疼痛,有效改善心腦血管微循環(huán)障礙和器官移植后抗栓治療,用以抑制血栓形成,防止移植后的器官和血管內(nèi)的血栓形成和血運(yùn)障礙[3]。
中國(guó)藥典、美國(guó)藥典、歐洲藥典、日本藥典和國(guó)家標(biāo)準(zhǔn)(國(guó)家食品藥品監(jiān)督管理局國(guó)家藥品標(biāo)準(zhǔn)WS1 -(X-041)-2002Z-2008)均未關(guān)注前列地爾注射液中可能由合成過(guò)程引入及貯存過(guò)程降解產(chǎn)生的各種雜質(zhì)[4-8]。因此,有必要建立一套專屬性強(qiáng),重復(fù)性好的有關(guān)物質(zhì)檢查法,有效控制前列地爾注射液的質(zhì)量。本研究參考《歐洲藥典》(EP10.1)中的前列地爾原料標(biāo)準(zhǔn)[6],建立前列地爾注射液有關(guān)物質(zhì)的檢查方法,并采用強(qiáng)制破壞試驗(yàn),驗(yàn)證主成分與已知雜質(zhì)及相鄰的未知雜質(zhì)之間達(dá)到基線分離,同時(shí)對(duì)已知雜質(zhì)進(jìn)行定量分析(校正因子雜質(zhì)D、E、F、G、H和單個(gè)雜質(zhì)的校正因子分別為1.0、0.7、0.8、0.7、0.7和1.0,來(lái)源于EP10.1),最終,采用加校正因子的主成分自身對(duì)照法,對(duì)前列地爾注射液進(jìn)行有關(guān)物質(zhì)檢查。
1.1 試藥和試劑
前列地爾對(duì)照品(LGC GmbH公司,批號(hào)77025,含量100.2%)、雜質(zhì)G對(duì)照品(EP,批號(hào)3,含量99.6%)、雜質(zhì)H對(duì)照品(EP,批號(hào)3,含量按100%計(jì))、前列地爾注射液(批號(hào)20190703,規(guī)格1 mL∶5 mg)和空白輔料溶液(批號(hào)19060601)均來(lái)自輔必成(上海)醫(yī)藥科技公司。