基于CuB(C_(6)H_(5))_(4)缔合物微粒体系的共振散射光谱法测定异烟肼药片中异烟肼的含量

Determination of Isoniazid in Isoniazid Drug Tablets by Resonance Scattering Spectrometry Based on CuB(C_(6)H_(5))_(4)Associaton Particles System

将异烟肼(INH)药片研碎后分取含2.00 mg INH的药粉,加入20 mL水,用玻棒搅拌2 min,所得溶液用蒸馏水稀释至100.0 mL,分取5.00 mL,离心2 min,分取上清液1.00 mL,用水稀释至10.0 mL,即制得INH样品溶液。取1.00 mL 0.20 mol·L^(-1)乙酸-乙酸钠缓冲溶液(pH 4.6)、1.50 mL 0.50 g·L^(-1)NaB(C_(6)H_(5))_(4)溶液、2.00 mL 1.00 g·L^(-1)CuSO4溶液,混匀后加入1.00 mL INH样品溶液,用水稀释至10.0 mL,振荡0.5 min,静置反应10 min,INH将Cu(Ⅱ)还原为Cu(Ⅰ),Cu(Ⅰ)与B(C_(6)H_(5))_(4)-反应生成CuB(C_(6)H_(5))_(4)缔合物,并进一步聚集生成缔合微粒,形成具有较强共振光散射信号的固液界面。分取适量上述溶液,置于荧光分光光度计中,在200~700 nm内同步扫描激发(λex)、发射(λ_(em))波长(λ_(em)=λex),读取397 nm处共振散射峰的信号强度IRSS。同步测定试剂空白,得到IRSS0,以ΔIRSS(ΔIRSS=IRSS-IRSS0)进行定量。结果显示:100倍淀粉、120倍葡萄糖、150倍麦芽糖、200倍蔗糖、600倍NO3-,800倍Cl-、1000倍K+、800倍Zn2+不干扰INH的测定;INH的质量浓度在0.010~2.00 mg·L^(-1)内和对应反应体系ΔIRSS呈线性关系,检出限(3s/k)为0.004 mg·L^(-1);方法用于实际样品分析,所得INH测定值的相对标准偏差(n=5)为1.8%~2.6%,测定值和《中华人民共和国药典》(2020年版)高效液相色谱法的基本一致;按照标准加入法进行回收试验,回收率为97.0%~99.0%。

Isoniazid(INH)drug tablets were ground,and the drug powder containing 2.00mg of INH was taken.After adding 20mL of water,the mixture was stirred for 2min with glass rod.The obtained solution was diluted to 100.0mL by water.An aliquot(5.00mL)was taken,and centrifuged for 2min,and 1.00mL of the supernatant was taken,and diluted to 10.0mL by water to prepare the INH sample solution.Then 1.00mL of 0.20mol·L^(-1)acetic acid-sodium acetate buffer solution(pH 4.6),1.50mL of 0.50g·L^(-1)NaB(C_(6)H_(5))_(4) solution,and 2.00mL of 1.00g·L^(-1)CuSO4solution were added and mixed,and 1.00mL of INH sample solution was added.The mixture was diluted to 10.0mL by water,shaken for 0.5min,and settled to react for 10min.INH reduced Cu(Ⅱ)to Cu(Ⅰ),and Cu(Ⅰ)reacted with B(C_(6)H_(5))_(4)-to form CuB(C_(6)H_(5))_(4) complex,which further aggregated to form associated particles,forming a solid-liquid interface with strong resonance light scattering signal.An appropriate amount of the above solution was taken and placed into a fluorescence spectrophotometer,and synchronously scan was made on excitation(λex)and emission(λ_(em))wavelengths in the range of 200-700nm(λex=λ_(em)),reading the signal intensity IRSSof the resonance scattering peak at 397nm.The reagent blank was analyzed simultaneously to obtain I0 RSS,withΔIRSS(ΔIRSS=IRSS-I0 RSS)for quantification.It was shown that 100times starch,120times glucose,150times maltose,200times sucrose,600times NO3-,800times Cl-,1000times K+,and 800times Zn2+did not interfere with the determination of INH.Linear relationship between values of the mass concentration of INH andΔIRSSof the reaction system was kept in the range of 0.010-2.00mg·L^(-1),with detection limit(3s/k)of 0.004mg·L^(-1).The proposed method was used for the analysis of actual samples,and RSDs(n=5)of the INH determined values were found in the range of 1.8%-2.6%.The determined values were basically consistent with those obtained from the high performance liquid chromatography in the Pharmacopoeia of the People′s Republic of China(2020version).Test for recovery was conducted according to the standard addition method,giving recoveries in the range of 97.0%-99.0%.