HPLC法测定注射用替曲朵辛盐酸利多卡因的含量及有关物质

HPLC determination of contents and related substances in the tetrodotoxin lidocaine hydrochloride for injection

目的建立注射用替曲朵辛盐酸利多卡因的含量及有关物质检测方法。方法采用高效液相色谱法,替曲朵辛的检测条件为:ZORBAX SB-C8柱(4.6mm×250mm,5μm),流动相为辛烷磺酸钠的磷酸盐缓冲液,流速0.3mL·min^(-1),激发波长365nm,发射波长510nm,柱后衍生剂为4mol·L^(-1)氢氧化钠溶液,衍生剂流速0.3mL·min^(-1),柱后衍生温度110℃,进样量50μL;盐酸利多卡因的检测条件为:TECHMaTe C18-STⅢ柱(4.6mm×250mm,5μm),流动相为磷酸盐缓冲液-乙腈(50∶50),流速1mL·min^(-1),柱温30℃,紫外检测波长254nm,进样量5μL。结果替曲朵辛和盐酸利多卡因的线性范围分别为0.102 4~10.24μg·mL^(-1)和0.501 0~15.032 4 mg·mL^(-1);平均回收率分别为100.48%和99.78%,RSD分别为1.13%和0.65%(n=9);检测限分别为0.41ng·mL^(-1)和10.3μg·mL^(-1),定量限分别为0.81ng·mL^(-1)和28.5μg·mL^(-1)。结论本方法可用于测定该制剂中替曲朵辛和盐酸利多卡因的含量及有关物质。

Objective To establish an HPLC method for determining the contents and related substances in the tetrodotoxin lidocaine hydrochloride for injection.Methods The analysis of tetrodotoxin was performed on a ZORBAX SB-C8column（4.6 mm×250 mm,5μm）,using a mobile phase of phosphate buffer solution of octane sulfonate at the flow rate of 0.3mL·min^（-1）.The excitation wavelength and emission wavelength were set at 356 and 510nm,respectively.The post column derivative reagent was 4mol·L^（-1） NaOH at the flow rate of 0.3 mL·min^（-1）,the post column derivatization temperature was maintained at 110 ℃ and injection volume was 50μL.The analysis of lidocaine hydrochloride was performed on a TECHMaTe C18-STII column（4.6mm×250mm,5μm）,using a mobile phase of phosphate buffer-acetonitrile（50∶50）at the flow rate of 1.0 mL·min^（-1）.The column temperature was maintained at 30℃.The detection wavelength was set at 254 nm and injection volume was 5μL.Results The linear ranges of tetrodotoxin and lidocaine hydrochloride were 0.102 4～10.24μg·mL^（-1） and0.501 0～15.032 4 mg· mL^（-1）,respectively.The average recoveries of tetrodotoxin and lidocaine hydrochloride were 100.48% and 99.78%,respectively.The RSDs of this method were all less than2%（n=9）.The limits of detection of tetrodotoxin and lidocaine hydrochloride were 0.41ng·mL^（-1）and 10.3μg·mL^（-1）,respectively.And the limits of quantitation of tetrodotoxin and lidocaine hydrochloride were 0.81ng·mL^（-1） and 28.5μg·mL^（-1）,respectively.Conclusion The method was applicable for the contents and related substances control of tetrodotoxin lidocaine hydrochloride for injection.