Using Computer Simulated Results of a Bulk Drug Substance Assay to Determine Acceptance Criteria for Method Validation

Purpose. To determine the statistical variability expected for a well designed HPLC assay of a bulk drug substance (BDS). The results are used to develop appropriate acceptance criteria for a method validation protocol as well as to evaluate the level of uncertainty expected for assay results using a variety of sampling/injection schemes. Methods. Computer simulation was used to generate a large quantity of data and the variability of the mock results was evaluated. Error propagation was also calculated, whenever possible, to confirm results obtained from the simulations. Results. Protocol acceptance criteria were developed that were consistent with the expected variability for data resulting from the execution of the validation protocol. In certain cases simulations provided the only avenue of obtaining results that could not otherwise be readily determined. Conclusions. Computer modeling can be used to obtain suitable acceptance criteria for validation results which are consistent with method variability. This is particularly significant in the case of linearity where it has been difficult to develop acceptance criteria based on anything other than analyst intuition and experience. Assay simulations clearly demonstrated that the variability expected for a typical BDS assay is large relative to the average specification range and therefore little insight about relative purity can be gained comparing individual passing assay results.     

Fumonisins as a possible contributory risk factor for primary liver cancer: A 3-year study of corn harvested in Haimen, China, by HPLC and ELISA

Employing HPLC fluorometry, gas-liquid chromatography (GLC) and a novel enzymelinked immunosorbent assay (ELISA) based on a monoclonal antibody, 40 corn samples, each collected in 1993 from agricultural stocks for human consumption in Haimen (Jiangsu County) and Penlai (Shandong Province), high- and low-risk areas for primary liver cancer (PLC) in China, respectively, were analysed for fumonisins (FBs), aflatoxins (AFs) and trichothecenes. Levels and positive rates of FBs and deoxynivalenol (DON) were significantly higher in Haimen than in Penlai. ELISA of the 40 corn samples harvested in the two areas in 1994 revealed that FB contamination levels and rates in these areas were comparable to those observed in 1993 in Haimen. ELISA analysis of 1993 and 1994 products revealed a wide occurrence of AFB₁ but the positive rates as well as levels were not significantly different between these areas. ELISA of the same sample number of corn harvested in 1995 revealed that FB contamination in Haimen was significantly higher than in Penlai. These 3-yearly surveys of corn samples (240 in total) demonstrated that corn harvested in Haimen was highly contaminated with FBs and that the contamination level, as well as positive rate in 1993 and 1995, were 10–50-fold higher than those in Penlai, suggesting FBs as a risk factor for promotion of PLC in endemic areas, along with the trichothecene DON. Co-contamination with AFs, potent hepatocarcinogens, was assumed to play an important role in the initiation of hepatocarcinogenesis.     

奈替米星体液浓度测定的方法学研究

通过对国产硫酸奈替米星注射后人体血尿中药物浓度测定，建立了奈替米星体液浓度测定的微生物法，该方法的检测限为０．０３７５～０．０７５１μｇ，最低检测浓度为０．２５～０．５μｇ／ｍｌ，回收率为８８．８％±３．６３％～９８％±１４．４０％，重复性试验相对标准差为２．２２％～５．３４％；并对１０例健康受试者用药后血标本１００余份进行了微生物法测定和高压液相色谱法测定，对两种方法测定结果的比较发现：微生物法测定的血药浓度明显高于ＨＰＬＣ法（Ｐ＜０．０１），而ＨＰＬＣ法由于需复杂的样品预处理和特殊的色谱条件，测定的血药浓度较低，回收率仅７１．９８％～７６．９６％。不过，两种方法测定结果的相关性较好。本研究结果提示：微生物法测定奈替米星体液浓度简便、易操作、结果准确可信，适用于体液药物浓度测定。     

涎腺腺癌细胞系对葫芦素BE的敏感性研究

目的 观察其有多种生物活性的葫芦素BE对涎腺腺癌细胞的杀伤作用。方法 用四唑盐显色法(MTT法)检测人腺样食性癌(ACC-M)和粘液表皮样癌(MEC-1)细胞系对葫芦素BE的敏感性实验。结果 葫芦素BE对两种腺癌细胞系均有较高的相对抗肿瘤活性(RAA值分别为345、148)。结论 实验结果为葫芦素BE治疗涎腺腺癌的进一步研究提供了重要依据。     

血清同工铁蛋白酶联免疫吸附测定及其临床观察

用酶联免疫吸附测定法(ELISA)观察了96例正常人,11例原发性肝细胞肝癌(HCC)、28例乳癌(BC)、31例肺癌(LC)、26例乳腺纤维瘤(BF)、11例肺炎(LF)及11例肺结核(TB)患者的血清同工铁蛋白(Serum isoferritin,SIF)水平。结果显示:正常人与HCC、BC、LC、BF及LF患者之间的SIF水平存在非常显著差异(P<0.01或P<0.001),在上述各类疾病中恶性肿瘤与相应组织的非癌性疾病患者之间的SIF水平也呈现非常显著的差异(BC与BF间、LC与LF间的P<0.01,LC与TB间的P<0.001)。SIF对检测HCC、LC及BC有较高的灵敏度,对鉴别这些恶性肿瘤具有重要价值。     

猪血清胸腺因子免疫活性的测定

根据哺乳动物循环血液中所含有的血清胸腺因子(STF)能使θ-阴性的玫瑰花瓣状形成细胞(RFC~-)转变成RFC~+,同时提高其对抗θ抗原血清的代用品硫唑嘌呤的敏感性的原理,采用成熟的BALB/C品系小鼠,经摘去胸腺后的鼠脾脏淋巴细胞和绵羊红细胞(SRBC)之间,在STF和硫唑嘌呤同时共存的条件下具有抑制玫瑰花结形成的作用,从而建立了对哺乳动物循环血液中的STF(九肽)的超微量生物活性测定方法。用本方法对猪血清和血浆的检测结果,其活性滴度分别为1:512、1:1024。     

多抗甲素对人NK细胞活性的作用机理研究

作者应用单细胞细胞毒试验及形态学观察，研究多抗甲素（ＰＡＡ）对人体自然杀伤（ＮＫ）细胞活性的作用机理。结果表明：ＰＡＡ能增强食道癌患者淋巴细胞（ＰＢＬ）明显低下的结合率（Ｂ％）和杀伤率（Ｋ％），也可增强正常人ＰＢＬ的Ｋ％，但对正常人ＰＢＬ的Ｂ％无影响。形态学观察发现：与靶细胞结合的ＰＢＬ除大颗粒ＰＢＬ外，尚有普通的非颗粒ＰＢＬ，结合的靶细胞损伤首先表现为线粒体肿胀、空泡变，然后核固缩，核膜损伤，细胞严重空泡变性等，偶见靶细胞与自然杀伤细胞结合部质膜破损的现象。经ＰＡＡ预处理的靶效细胞结合的形态改变无明显差异，但食道癌组的杀伤作用出现较早，且明显增强。     

造血系统恶性肿瘤患者血清sIL—2R检测的临床意义

使用ＥＬＩＳＡ双抗夹心法对临床４３例造血系统恶性肿瘤患者及２０例健康体检人员的血清ｓＩＬ－２Ｒ进行检测，并对ＡＬＬ和ＡＮＬＬ复发与未复发患者的血清ｓＩＬ－２Ｒ水平进行了比较分析。实验结果表明，血清ｓＩＬ－２Ｒ水平的升高是造血系统恶性肿瘤的重要标志之一；测定血清ｓＩＬ－２Ｒ水平对造血系统恶性肿瘤的辅助诊断、疗效观察及预后估计具有重要意义。     

肝病患者肝组织中丙型肝炎病毒感染的初步研究

采用直接酶标记法对174例慢性肝炎、重症肝炎和肝癌组织中的HCV抗原进行了检测,HCV 抗原的检出率分别为10.71％、8.10％和2.68％。表明在我国慢性和重症肝病中存在HCV的感染。通过对HCV与HBV 感染关系的研究,发现肝内HBV 标志阳性和阴性者HCV抗原的检出率无明显差异,说明HCV感染与HBV感染之间似无明显关系。     

Increased in vitro uptake of the complement C3b in the serum of patients with Guillain-Barré syndrome, myasthenia gravis and dermatomyositis

To examine the role of complement in certain autoimmune neuromuscular diseases, we used an in-vitro quantitative complement uptake assay that allows measurement of the capacity of patients' sera to deposit fragments of the third complement component onto sensitized targets. C3 uptake was significantly higher in patients with active dermatomyositis, Guillain-Barré syndrome and myasthenia gravis, compared to inclusion body myositis and controls. The in-vitro C3 uptake assay supports the role of C3b neoantigen and Membranolytic Attack Complex deposition in the target tissues and may be a useful tool to monitor disease activity in patients with complement-mediated neurological disorders.