HPLC法同时测定氯霉素滴眼液中有关物质和尼泊金酯类防腐剂

HPLC determination for related substances and antimicrobial agents content of chloramphenicol eye drops

目的:建立高效液相色谱法同时测定氯霉素滴眼液中有关物质(氯霉素二醇物和对硝基苯甲醛)和防腐剂(对羟基苯甲酸甲酯、乙酯和丙酯)的含量。方法:用十八烷基硅烷键合硅胶为填充剂;流动相为水(含0.1mol·L-1庚烷磺酸钠溶液,0.5mol·L-1磷酸二氢钾与0.5%三乙胺,混匀,用磷酸调pH至2.5)(A)-甲醇(B),梯度洗脱[0min(68%A)→15min(68%A)→40min(35%A)→50min(68%A)→60min(68%A)];流速为1.0mL·min-1;检测波长为277nm。结果:二醇物、对硝基苯甲醛及对羟基苯甲酸甲酯、乙酯和丙酯的线性范围分别为3.56～14.24μg·mL-1(r=0.9999),1.74～6.96μg·mL-1(r=0.9999),9.20～36.80μg·mL-1(r=0.9998),8.56～34.24μg·mL-1(r=0.9999),4.80～19.20μg·mL-1(r=0.9999)。结论:本文建立的高效液相色谱法结果准确可靠,可作为氯霉素滴眼液中有关物质和防腐剂含量的控制方法。

Objective：To establish an RP-HPLC method for determination of related substances and antimicrobial agents content of chloramphenicol eye drops.Methods：ODS column was adopted,the mobile phase consisted of water（including 0.1 mol·L-1 1-heptanesulfonic acid sodium salt,0.5 mol·L-1 patassiumdihydrogen and 0.5% triethylamine,adjust pH=2.5 with phosphoric acid）（A）-methanol（B）with gradient elution[0 min（68%A）→15 min（68%A） →40 min（35%A）→50 min（68%A） →60 min（68%A）] at the flow rate of 1.0 mL·min-1 and the detection wavelength was 277 nm.Results：The linear response ranges of glycols,4-nitrobenzaldehyde,and methyl p-hydroxybenzoate,ethyl p-hydroxybenzoate and propyl p-hydroxybenzoate were 3.56-14.24 μg·mL-1（r=0.9999）,1.74-6.96 μg·mL-1（r=0.9999）,9.20-36.80 μg·mL-1 （r=0.9998）,8.56-34.24 μg·mL-1 （r=0.9999）and 4.80-19.20 μg·mL-1 （r=0.9999）.Conclusion：The result of determination is accurate and reliable.This method is suitable for determination of related substances and antimicrobial agents content of chloramphenicol eye drops.