新型镇痛多肽Eb1.6的含量及有关物质检测

Determination of content of novel analgesic conopeptide Eb1.6 and its related substances

目的:建立一种适用于新型alpha芋螺镇痛多肽Eb1.6含量及有关物质的分析方法,以及其水分和乙酸的含量测定方法。方法:采用高效液相色谱法测定Eb1.6原料药中Eb1.6的含量。使用Zorbax XDB-C18色谱柱(150 mm×4.6 mm,5μm),流动相A为含0.1%三氟乙酸(TFA)的水,流动相B为含0.1%TFA的乙腈,梯度洗脱,流速1 m L·min-1,检测波长214 nm,柱温25℃;采用卡尔菲休库仑滴定法及高效液相色谱法测定样品中水分及乙酸含量。结果:3批Eb1.6原料药中Eb1.6含量范围在(98.85±0.63)%^(99.04±0.33)%之间(按无水物计算),总有关物质含量在0.45%~0.84%之间,样品中水分含量在(1.23±0.32)%^(2.23±0.23)%之间,乙酸含量在(11.16±0.08)%^(12.66±0.03)%之间。结论:该方法快速简便,结果准确可靠,可用于Eb1.6的质量分析。

Objective： To established a method to determine the content of novel analgesic conopeptide Ebl. 6 and its related substances as well as the content of moisture and acetic acid. Methods： Ebl. 6 was detected with Zorbax Eclipse XDB - C18 column（ 150 mm×4.6 mm,5 μm） at the detection wavelength of 214 nm with the flow rate of 1.0 mL · min^- 1. The mobile phase A was 0. 1% TFA, the mobile phase B was 0. 1% CH3 CN, the gradient elution was adopted. The column temperature was 25 ℃C. The water content of Ebl. 6 was determined by Karl Fischer coulo- metric titration method, and the content of acetic acid was determined by HPLC method. Results： The content of Ebl. 6 in three batches of crude drugs was in the range of（98.85 ± 0. 63） % - （99.04 ± 0. 33 ） % （ calculated by anhydride）, while the content of total related substances was from 0.45 % to 0. 84%, the water content was （ 1.23 ±0.32） % - （2.23 ± 0. 23 ） % and the content of acetic acid was（ 11.16 ± 0. 08） % - （ 12.66± 0.03 ） %. Conclusion： The method is reliable, accurate and specific. It can be used for quality analysis of Ebl. 6.