比对法在计量标准考核验证中的应用探讨

为探讨比对法在计量标准考核验证中的可行性,将5家实验室分别对同一分度吸量管的检定结果进行比对,同时采用具有溯源性的传递比较法进一步验证比对法的可靠性。验证结果显示,采用比对法和传递比较法对被考核实验室的验证结果一致,说明采用比对法对检定结果的验证是有效的、可行的。     

葡萄糖氧化酶的固定化方法与改进

本文在固定化葡葡糖氧化酶方面,对几种固定化载体和方法做了最佳条件摸索和比较。改进了戊二醛偶联法,尝试用二乙醇胺先保护戊二醛一端,另一端与载体偶联,然后脱去保护基再偶联酶,效果较好。用该法制得固定化酶在所研究的三种方法中效果最好。固定化酶活力回收率比用聚丙烯酰胺包埋法高出一倍多,比用重氮盐共价键合法高出10%左右。     

鱼类干货中过氧化氢(H2O2)残留的测定

利用过氧化氢与高锰酸钾发生定量氧化还原反应引起高锰酸钾溶液的吸光度下降,建立了用紫外-可见分光光度计测定鱼类干货(鱼鳞、鱼皮)中微量过氧化氢的方法。结果表明,回收率在87.7%~105.4%之间,该法可测定鱼类干货中的微量过氧化氢,方法简单、快速、准确度高。     

间二氯苯纯度的测定

通过测定间二氯苯中的杂质含量得到间二氯苯的纯度。分别采用高效液相色谱法(HPLC–UVD)和气相色谱法(GC–FID)测定间二氯苯中主要杂质邻、对二氯苯的含量,结果表明两种方法所得杂质含量一致;用高效液相色谱法(HPLC)测定间二氯苯中杂质苯的含量,卡尔费休(Karl Fischer)法测定水分含量,热重法(TGA)进行灰分测定,电感耦合等离子体质谱(ICP–MS)法测定无机杂质含量。最终确定间二氯苯的纯度为99.60%,扩展不确定度为0.03%(k=2)。该方法测定结果准确可靠,具有可溯源性。     

分段流动分析法测定海水中挥发酚

研究了分段流动分析法测定海水中痕量挥发酚。在实验优化的最佳条件下测定海水中挥发酚,与国家标准方法有良好的相关性。方法的相对标准偏差(RSD)为1.9%,略优于国家标准方法,且操作简便。应用于海水中挥发酚的分析,取得结果满意。     

全自动凯氏定氮仪测定化肥中的含氮量

建立全自动凯氏定氮仪检测化肥中氮含量的分析方法。称取总氮量在0.03~0.3 g范围的样品,经消解仪消解及全自动凯氏定氮仪碱化蒸馏,以硼酸为接收液,用0.15~0.20 mol/L的盐酸标准溶液进行自动滴定。该方法加标回收率为99.0%~101.5%,测定结果的相对标准偏差小于1.0%(n=6)。与国标GB/T 8527–2010法测定结果无显著性差异,全自动凯式定氮仪法的准确度、精密度均优于国标方法,而且所用试剂少、分析时间短,满足实验室快速检测大批量化肥中含氮量的需要。     

介孔分子筛SBA-15的脂肪酶固定量分析测定

利用吸附法将假丝酵母脂肪酶(candida rugosa lipase,CRL)固定在介孔分子筛SBA-15上,对比了由单波长紫外分光光度法、双波长紫外分光光度法和二辛可宁酸法(bicinchoninic acid method, BCA)法测定的酶蛋白浓度及酶蛋白固定量.结果表明: SBA-15对紫外吸收有明显干扰,单波长紫外法测定结果远大于双波长紫外法和BCA法,双波长紫外法和BCA法测定结果较接近.利用BCA法测定了不同浓度CRL在介孔分子筛上的固定量,考察了固定化酶的泄漏量.在编号分别为Lu001和LLSD1的介孔分子筛SBA-15上的载酶量分别为16.6和114.12 mg/g.在缓冲溶液中SBA-15固定化酶的泄漏率只约为0.5%,可作为良好的酶固定化载体.     

凝胶色谱法测定聚丙烯腈相对分子质量的验证

采用窄分布的聚苯乙烯和聚甲基丙烯酸甲酯分别对凝胶色谱柱进行标定,测定了聚丙烯腈的相对分子质量。聚苯乙烯、聚甲基丙烯酸甲酯标定法得到聚丙烯腈的重均相对分子质量分别为5.008×10^5,2.929×10^5,两种标定方法所得数据相差较大。采用光散射法验证窄分布标准样品标定测试数据的准确度,得到聚丙烯腈的重均相对分子质量为1.485×10^5,与窄分布聚合物标定凝胶色谱法得到的数据相差很大,表明凝胶色谱标定法测定聚丙烯腈的相对分子质量是一种相对方法,其量值无法实现准确溯源。     

磷酸浸渍黏土砖中五氧化二磷含量测定方法的改进

研究了磷钼酸铵容量法测定磷酸浸渍黏土砖中五氧化二磷含量的方法,对国家标准分析方法进行了改进,讨论了不同实验条件对实验结果的影响,并进行了精密度和准确度实验。结果准确、重现性好,证明方法可靠。     

Liquid chromatographic determination of pyrantel tartrate in medicated formulations

The validation of a novel liquid chromatographic (LC) method for the determination of pyrantel tartrate in feed is presented. The method provides a significant improvement over the efficiency and precision of AOAC Official Method 978.30. The method was shown to be accurate, precise, linear, and robust for medicated articles. Unlike the official method, the LC method was shown to be a superior stability-indicating method. After the method was validated by using laboratory blends, the effectiveness of the method was demonstrated with marketed product as well.     

固相合成胸腺五肽(TP5)

采用Fmoc固相多肽合成中的活化酯方法和2,6-二氯苯甲酰氯(DCB)混合酸酐法, 对Fmoc-Tyr(t-Bu)-OH与Wang树脂反应中的反应级数和表观活化能进行了研究, 并采用常规方法和微波强化方法分别进行了胸腺五肽的合成. 实验结果表明, 活化酯方法的反应级数为1.855, 表观活化能15.24 kJ/mol, 混合酸酐法的表观活化能为35.14 kJ/mol. 与传统方法相比, 微波将缩合反应速率提高了30倍以上, 氨基酸过量倍数也从传统的三倍降低到两倍.     

新型薄层色谱内标法测定葛根素含量

以大豆甙元为内标物在高效硅胶薄层板上采用新型内标法测定了葛根素注射液、中药葛根及中成药玉泉丸中葛根素的含量,建立了新型TLC内标法测定葛根素的新方法。结果表明：用大豆甙元作为内标物测定葛根素可以满足TLC内标物的一般条件,新型TLC内标法测定葛根素的含量具有测定结果准确,精密度好,回收率高等优点。     

New refractive index method for measurement of alcoholic strength of small volume samples

A study of the correlation between the refractive index (RI) method adopted by the European Community (EC) and a reference pycnometric method for the measurement of alcoholic strength was undertaken. A new RI method with greater accuracy was also developed. Alcoholic strength measured by both RI methods presented a relatively constant negative bias compared with results by the pycnometric method. Differences found between the RI methods and the pycnometric method were 0.6-0.9% (v/v) when RI was measured by the EC method and 0.4-0.5% (v/v) when the new RI method developed in our laboratory was used. Statistical analysis of the results showed that differences between the 2 RI methods and the reference pycnometric method were statistically significant at the 95% confidence level. Correction factors are proposed for the accurate use of measurements of alcoholic strength obtained for small volume samples.     

t检验法用于评价中药色谱指纹图谱的相似度

本文采用t检验法评价中药色谱指纹图谱的相似度。利用来自六个不同厂家的三黄片样品的高效液相色谱指纹图谱数据来验证t检验法的适用性,并用主成分分析(PCA)进一步验证方法的可行性。在PCA的聚类图上,t检验无显著性差异的样品聚为一类,t检验有显著性差异的样品各为一类。结果表明:用t检验法能客观地反映中药样品间的相似性和差异性。     

甲氧氯普胺的流动注射光度分析

基于在盐酸介质中,利用甲氧氯普胺－亚硝酸钠－α－萘胺的重氮化－偶合反应,建立了流动注射光度法测定甲氯普胺的新方法,利用单纯形优化法选择了最佳实验条件。方法的线性范围为0.0-30mg/L,进样频率为160／h。11次重复测定的相对标准偏差小于0.6％,该方法已成功用于药物中甲氧氯普胺含量的测定,测定结果与药典法对照,并经统计学处理,结果满意。     

Determination of rosuvastatin in the presence of its degradation products by a stability-indicating LC method

A forced degradation study was successfully applied for the development of a stability-indicating assay method for determination of rosuvastatin Ca in the presence of its degradation products. The method was developed and optimized by analyzing the forcefully degraded samples. Degradation of the drug was done at various pH values. Moreover, the drug was degraded under oxidative, photolytic, and thermal stress conditions. Mass balance between assay values of degraded samples and generated impurities was found to be satisfactory. The proposed method was able to resolve all of the possible degradation products formed during the stress study. The developed method was successfully applied for an accelerated stability study of the tablet formulation. The major impurities generated during the accelerated stability study of the tablet formulation were matches with those of the forced degradation study. The developed method was validated for determination of rosuvastatin Ca, and the method was found to be equally applicable to study the impurities formed during routine and forced degradation of rosuvastatin Ca.     

离子色谱测定烟草中果胶方法的改进

为了提高YC/T 346–2010果胶含量测定方法的分析速度,将标准中醇化除糖、滤渣酸化两步前处理优化为一步,即酸醇溶液加热回流同时除糖和酸化,并将方法改进前后的测定结果进行比较分析。一步前处理方法线性相关系数为0.999 9,回收率为93.57%~102.29%,测定结果的相对标准偏差为4.92%(n=5)。该方法简化和缩短了样品前处理步骤和时间,测定结果与YC/T 346–2010一致,可用于烟草中果胶含量的批量测定。     

Determination of Atazanavir in the Presence of its Degradation Products by a Stability-Indicating LC Method

A forced degradation study was successfully applied for the development of a stability-indicating assay method for the determination of atazanavir in presence of its degradation products. The method was developed and optimized by analyzing the forcefully degraded samples. Degradation of the drug was done under acidic, alkaline, oxidative, photolytic and thermal stress conditions. The proposed method was able to resolve all of the possible degradation products formed during the stress studies. The major impurities are generated in acidic and alkaline conditions. The product was found stable under thermal, photolytic and oxidative conditions. The developed method was validated for determination of atazanavir, and the method was found to be equality applicable to study the impurities formed during routine and forced degradation of atazanavir.     

Determination of creatinine in serum by high-performance liquid chromatography: a comparison of three ion-exchange methods

Three ion-exchange high-performance liquid chromatographic methods for the determination of creatinine in serum have been compared. In method 1 a strong cation exchanger was used. In method 2 a reversed-phase column was given strong cation-exchange properties by the addition of N-methyloleoyl taurate to the mobile phase. In method 3 a weak cation exchanger was used. Elution was with a pH gradient in methods 1 and 2, and isocratic elution was used in method 3. The imprecision was similar for the three methods and varied between 0.9 and 2.5% as studied within-day and between 1.4 and 3.2% from day-to-day. The lowest coefficient of variation was obtained around the upper reference limit. Analytical recoveries were quantitative for the three methods. The method with N-methyloleoyl taurate showed no advantages over the conventional strong cation exchanger. With the weak cation exchanger no interferences were detected from compounds investigated, but with the strong cation exchanger a slight interference was obtained with uric acid. We prefer the weak cation-exchange method because of its simplicity, higher throughput and absence of interference from hitherto tested compounds.     

气相色谱测定牦牛骨中脂肪酸的甲酯化方法比较

利用气相色谱（GC）技术,采用酸水解提取脂质,比较了6种甲酯化法（乙酰氯-甲醇法、H₂SO₄-甲醇法、HCl-甲醇法、KOH-甲醇法、KOH-甲醇+H₂SO₄-甲醇法和KOH-甲醇+HCl-甲醇法）对脂肪酸测定的影响,优选牦牛骨中脂肪酸测定的最佳方法。37种脂肪酸标准样品在0.28~250.00 mg/L范围内线性关系良好,相关系数均大于0.99（除C4：0外）。碱酯化法和酸碱结合法几乎无法测出牦牛骨中的脂肪酸,其测得的总脂肪酸含量小于0.20 g/100 g。乙酰氯-甲醇法测得的总脂肪酸含量（13.61 g/100 g）显著高于H₂SO₄-甲醇法（总脂肪酸含量为11.68 g/100 g）和HCl-甲醇法（总脂肪酸含量为3.18 g/100 g）测得的结果。乙酰氯-甲醇法和H₂SO₄-甲醇法的日内和日间精密度分别为0.27%~8.60%和0.34%~2.64%,两种方法中脂肪酸的回收率为83.06%~105.54%。结果表明,酸水解-乙酰氯-甲醇法是牦牛骨中脂肪酸测定的最佳方法。C18：1n9c、C16：0、C18：0和共轭亚油酸（CLA）是牦牛骨的主要脂肪酸,其总和达脂肪酸总量的85%以上,饱和脂肪酸与不饱和脂肪酸含量比值约为1：2。牦牛骨中脂肪酸的研究为骨资源脂质的有效利用提供了重要依据。