抗生素类药品无菌检查方法研究进展

无菌检查法是无菌药品检查的法定检验方法,其中薄膜过滤法因易操作、风险低和准确度高等优点比直接接种法应用更广泛。无菌药品剂型种类繁多,其中抗生素类药品因品种结构多变、抑菌性强和抗菌谱广等因素,一直以来是建立无菌检查方法的难点。本文就薄膜过滤法中的溶解、过滤、冲洗和培养等4个关键步骤展开讨论,通过对已建立的13类47个品种共61个抗生素药品无菌检查方法的综述,探讨建立无菌检查方法的基本策略,并对无菌检查方法和结果的局限性进行讨论,以期为建立抗生素类无菌药品无菌检查方法及正确认识无菌检查结果提供思路和参考。     

疫苗类制品无菌检查方法(薄膜过滤法)的研究

目的:疫苗制品的无菌检查方法中薄膜过滤法的建立和方法学验证。方法:选取5个企业21个品种的疫苗制品,采用薄膜过滤法进行无菌检查,选取金黄色葡萄球菌、枯草芽孢杆菌、大肠埃希菌、生孢梭菌、白色念珠菌和黑曲霉作为验证菌进行方法学验证。结果:实验结果表明,疫苗类生物制品对真菌的抑菌作用较细菌更弱,疫苗类生物制品的敏感菌株为枯草芽孢杆菌,含防腐剂硫柳汞的疫苗的抑菌作用较其他类的疫苗更强。采用薄膜过滤法进行试验时,某些疫苗样品由于含氢氧化铝佐剂而不易通过薄膜,可将稀释液更换为磷酸盐缓冲液(pH 6.0),可使其顺利通过滤膜。结论:按照不同疫苗制品的特性,将其大致分为三类,并分别建立了无菌检查法。     

不同剂型硝酸咪康唑微生物限度检查方法适用性研究与评价

目的通过不同的前处理方式,建立硝酸咪康唑不同剂型的微生物限度检查法。方法参照《中国药典》2015年版四部,根据硝酸咪康唑胶囊剂、散剂和乳膏剂的理化特点,选择均质、萃取和静置等前处理方式,采用平皿法和薄膜过滤法建立适用于硝酸咪康唑不同制剂的微生物限度检查方法,并比较方法差异。结果建立的3种不同剂型硝酸咪康唑的微生物限度检查方法满足《中国药典》2015年版要求,需氧菌总数,霉菌和酵母菌总数检查法中的试验菌回收率均在50%~200%之间,加入的控制菌均可检出,各方法适用于不同剂型硝酸咪康唑的微生物限度检查。结论硝酸咪康唑不同剂型的微生物限度检查方法差异较大,应灵活选择适宜的前处理方式,解决样品抑菌性的去除和难于过滤的问题。     

不同厂家生产的丹参注射液无菌检查方法验证

目的：建立不同厂家生产的丹参注射液无菌检查方法,保证无菌检查结果的准确性和可靠性。方法本试验按中国药典2010年版一部（附录XⅢB）所载“无菌检查法”项下进行。结果采用直接接种法对人工污染的六种菌株进行试验,不同厂家生产的丹参注射液对金黄色葡萄球菌都有不同程度的抑制作用。本方法是采用薄膜过滤法用0．9％的无菌氯化钠50mL进行样品稀释,后用不同冲洗量的0．1％蛋白胨水溶液分次冲洗,可消除抑菌成份。结论不同厂家生产的相同品种、相同规格的丹参注射液其无菌检查方法验证结果不同。     

头孢菌素类无菌检查中酶处理探讨

目的：通过验证试验建立头孢菌素类无菌检查法。方法：按2005年版《中国药典》规定，采用薄膜过滤法对10种不同代注射用头孢菌素进行系统的无菌检查方法研究。结果：不同代头孢菌素因抗菌谱的差异，对阳性对照菌，冲洗量及青霉素酶的加入方法等条件有显著不同。结论：头孢菌素类药的无菌检查方法可加入适量β-内酰胺酶（青霉素酶），提高阳性检出率，确保无菌实验结果的可信度。     

抗菌药物无菌检查法最佳冲洗条件选择

目的:选择抗菌药物无菌检查法的最佳冲洗条件.方法:采用中国药典2000年版二部附录XIH无菌检查法项下薄膜过滤法.结果:确定不同品种的抗菌药物最佳冲洗条件.结论:为不同品种抗菌药物无菌检查的具体冲洗量提供了依据,对药品生产及检验部门有实用价值.     

碱性成纤维细胞生长因子凝胶的无菌检查

目的建立碱性成纤维细胞生长因子（bFGF）凝胶剂的无菌检查方法。方法依据《中国药典》2010年版三部要求,采用直接接种法进行bFGF凝胶剂无菌检查并进行方法学验证。结果与结论通过方法学验证说明建立的无菌检查法适用于bFGF凝胶剂的无菌检查。     

双黄连注射液无菌检查的方法验证

目的:主要是建立一种适合于双黄连注射液进行无菌检查的方法.方法:薄膜过滤法与直接接种法.结果:可照薄膜过滤检查法的检查条件进行双黄连注射液的无菌检查.     

中国药典2000年版无菌检查法取样量与结果判定的探讨

中国药典2000年版规定:无菌检查法应用于无菌或灭菌制品、敷料、缝合线、无菌器具及其他应作无菌检查的品种,按照无菌检查范围检查结果为无菌时,由于它既受抽检样本数量的限制,又受生产和灭菌工艺的限制,所以在一定意义上讲,它表示的是最终灭菌品种达到了10~(-6)微生物存活概率的相对意义。     

注射用全氟丙烷人血白蛋白微球无菌检查方法验证

目的 建立适合于注射用全氟丙烷人血白蛋白微球无菌检查方法.方法 采用《中国药典》2015年版四部无菌检查法,分别采用薄膜过滤法和直接接种法对注射用全氟丙烷人血白蛋白微球进行无菌检查方法研究.结果 薄膜过滤法无法过滤供试品,因此采用直接接种法进行无菌检查.结论 直接接种法可用于注射用全氟丙烷人血白蛋白微球无菌检查.     

Solution stability of cephradine neutralized with arginine or sodium bicarbonate

The solution stability of two formulations of cephradine--one using L-arginine and the other sodium carbonate as the neutralizer--was studied. Solutions of each formulation of 1% cephradine were prepared in the following diluents: 0.9% sodium chloride injection, lactated Ringer's injection, Ringer's injection, Normosol-R injection, 5% dextrose injection, and sterile water for injection; 5 and 25% solutions were made with sterile water for injection. All solutions were maintained at 25 degrees C, and at least five samples of each were assayed at various time intervals. Assay methods were HPLC, hydroxylamine colorimetric assay, microbiological agar diffusion, and iodometric analysis. By all assay methods, degradation rates of 1% solutions were lower for the arginine-neutralized product than for the one neutralized with sodium carbonate. This may be attributable to the lower pH values of solutions of the formulation with arginine, because one mechanism of degradation is pH-dependent. At concentrations of 5%, the difference in cephradine stability between the two formulations was minimal. At the 25% concentration, the formulations containing sodium carbonate were more stable. At these higher concentrations, the effect of pH is less important because degradation occurs by a combination of mechanisms. The 1% cephradine-arginine formulation was more stable than the same strength cephradine-sodium carbonate formulation in all the i.v. diluents studied. At 5 and 25% cephradine concentrations, the differences in stability between the two formulations were not substantial.     

Formulation and evaluation of an effective pH balanced topical antimicrobial product containing tea tree oil

The effect of pH on the antimicrobial activity of Melaleuca alternifolia essential oil formulations was studied. Microemulsions, liposomal dispersions, multiple emulsions and a colloidal bed of sterile clay were formulated using 5% w/w of tea tree oil. A number of formulations were prepared at various pH values (5.0, 5.5, 6.0, 6.5, and 7.0). Thermal stability studies showed that the formulations were stable for more than eight months. Agar dilution tests showed MICs of 1.0% v/v S. aureus and S. epidermidis. In the broth dilution test, MBC of the oil for P. acnes was 0.5% v/v. MIC and MBC values were comparable to those of non-formulated tea tree oil, indicating that tea tree oil retained its activity in the above-mentioned formulations. The microbiological evaluation showed that the formulations containing 5% w/w tea tree oil had a maximum effect at pH 5.5.     

《中国药典》2020年版（一部）中收载含罂粟壳成方制剂的汇总与分析

目的 拓展含罂粟壳成方制剂的中医临床用药范围,规范含罂粟壳成方制剂使用说明与质量标准。方法 汇总《中国药典》2020年版（一部）（以下简称《中国药典》一部）中收载的含罂粟壳成方制剂,重点对含罂粟壳成方制剂的剂型、用法用量、使用禁忌、检验方法、处方中罂粟壳使用量、处方中罂粟壳的炮制方法等进行分析,提出了提高其临床安全用药水平的建议。结果 经对《中国药典》一部中收载的1 607个中药成方制剂处方项下的处方药味全部品种进行汇总统计分析,共有19个含有罂粟壳（含罂粟壳浸膏1个）的中药成方制剂,仅占《中国药典》（一部）全部中药成方制剂的1.24%。含罂粟壳成方制剂的基础研究比较薄弱,存在制剂品种少,个别品种用量不明确以及使用禁忌表述不规范等问题。结论 建议加强含罂粟壳成方制剂的基础研究,扩大临床用药范围,加强上市后安全性再评价,以达到规范使用,提高临床合理使用水平,保障含罂粟壳成方制剂的临床用药安全目的。     

A two-stage reverse dialysis in vitro dissolution testing method for passive targeted liposomes

A novel two-stage reverse dialysis method has been developed for in vitro release testing of liposomal drug product with passive targeting characteristics. The first stage of the test is to mimic the circulation of liposomes in the body, whereas the second stage is to imitate the drug release process at the target. Buffer and surfactant solution were used during the first and second stages, respectively. For formulations containing high phase transition temperature lipids and high cholesterol content, no drug leakage was observed during the first stage of test. In the second stage, however, formulations with different compositions showed significant differences in terms of drug release rate, and discriminatory ability of the method was demonstrated. On comparing two different membrane diffusion techniques, dialysis and reverse dialysis methods, the reverse dialysis method showed significantly lower variation, and therefore is the preferred method. The developed in vitro release testing method should help to distinguish formulations with varied compositions for quality control testing purposes. This two-stage reverse dialysis method may pave the way to the development of more bio-relevant release testing methods for liposomal drug products.     

两种除菌级疏水性过滤器完整性测试方法的比较

目的：比较两种除菌级疏水性过滤器完整性测试方法,提出适用性的建议及需要把握的要点,旨在为同行开展此类工作提供参考。方法：阐述无菌制剂生产过程中除菌级过滤器的法规要求,较全面地比较起泡点测试法与水侵入测试法的基本原理、影响因素及其在应用中的优劣性。结果：两种方法基于相同测试原理,适用限制条件略有不同,均能有效保证除菌过滤系统的完整性,降低制药质量风险。结论：水侵入测试法不需要引入有机溶剂等低表面张力溶液进行润湿,操作简便,更适宜于医院无菌制剂生产过程除菌级疏水性过滤器的完整性测试。     

In vitro and in vivo bactericidal activities of 10%, 2.5%, and 1% povidone-iodine solution

The bactericidal action of three formulations of a povidone-iodine (PVI) complex in vitro, in vivo, and in the presence of competing organic matter was evaluated. Bacterial organisms included Staphylococcus aureus ATCC 25923 and 25 clinical isolates of Staph. aureus, designated KU 1-25. For the in vitro studies, 1.0 mL of bacterial inoculum containing 10(7) organisms was introduced into 9.0 mL of chemically stable 10% and 1% PVI formulations in sterile culture tubes, and 1.0-mL samples were withdrawn at set intervals. Samples were plated by using standard techniques and incubated for 24 hours, after which colony-forming units were counted. For in vivo studies, 0.1 mL of 10(6) Staph. aureus ATCC 25923 or KU inoculum was deposited on the dorsum of the hand of healthy human subjects. This area was wiped with a cotton swab saturated with 1%, 2.5%, or 10% PVI formulations. Samples were taken at 15 and 30 seconds after application of the iodophor. To test the bactericidal activity of the three formulations in the presence of a competing substrate, a swab soaked with sterile sheep's blood was applied to the skin and allowed to dry. The percentage of 10(4) Staph. aureus inoculum recovered allowed for comparison of the three products. In vitro, the 1% PVI formulation was bactericidal for 10(7) Staph. aureus within two minutes, as compared with the four minutes required by 10% PVI. On the skin contaminated with 10(6) organisms, the rates of killing within 30 seconds were comparable for both solutions.(ABSTRACT TRUNCATED AT 250 WORDS)     

Dissolution properties of different designed and formulated salbutamol tablet dosage forms

Pharmaceutical availability or in vitro availability is one of the aspects of drug bioavailability. Dissolution can be described best as a tool that can provide valuable information about the availability of a drug product. Dissolution test was performed on three different designed and formulated of salbutamol tablet formulations marketed in Turkey. The test methods were the paddle method and the rotating basket method described in United States Pharmacopeia. All studied formulations showed a good agreement with pharmacopeial requirements. In particular all studied commercial tablet formulations showed a quite fast release of the antiasthmatic drug. Other type of table formulations showed slow release. In order to evaluate the dissolution rates five different kinetics have been examined and the best fitting kinetics was found to be RRSBW kinetic.     

布洛芬不同剂型微生物限度检查方法验证汇总

目的研究布洛芬在不同剂型中微生物限度检查方法。方法采用平皿法、培养基稀释法、薄膜过滤法对几种不同剂型布洛芬进行方法验证。结果混悬液、颗粒、片剂、胶囊剂型采用平皿法、稀释法即可,软胶囊及缓释片需采用薄膜过滤法进行微生物限度检查。结论布洛芬各剂型除软胶囊和缓释片有较强的抑菌性,其它剂型的抑菌性不大。     

Microemulsions with timolol as potential eye drops

Stable microemulsions oil-in-water (o/w) and water-in-oil (w/o) with timolol were obtained. These formulations were sterilized using two methods: by autoclaving and by sterile filtering according to the Polish Pharmacopeia V. Microemulsions were kept in the temperatures of 25 degrees C and 5 degrees C during three months. The following physical parameters were investigated: density, dynamic viscosity, refractive index, surface tension, pH and osmotic tension. All the microemulsions met the requirements for eye drops.     

Vehicle effects on in vitro transdermal absorption of sevoflurane in the bullfrog, Rana catesbeiana

The experimental objectives were to identify a vehicle which produces a homogenous formulation when combined with the anesthetic solution sevoflurane and understand the dermal absorption of sevoflurane in silastic membranes and amphibian skin in vitro utilizing a flow-through diffusion system. Seven vehicles were evaluated in varying ratios with 5 formulations resulting in the desired homogenous consistency for practical application. Sevoflurane diffusion across silastic membranes was influenced by pluronic/lecithin organogel (PLO), pluronic F 127 20% gel, and sterile lube. Flux and permeability across silastic membranes were significantly greater in sterile lube than in the other formulations. While no significant vehicle effects were observed in bullfrog skin, the flux-time profiles suggest that sevoflurane diffusion in bullfrog skin may be positively influenced by PLO. Future in vivo studies are required to assess sevoflurane retention after removal of these formulations to more accurately control the plane of anesthesia in amphibians.