酸溶辅助超声波法提取类球红细菌类胡萝卜素条件优化

通过单因素和正交试验,对类球红细菌3757产类胡萝卜素的提取条件进行了研究。先采用超声波法、酸溶法、研磨法、冻融法、酸溶辅助超声波法和冻融辅助超声波法优化了从类球红细菌3757菌株中提取类胡萝卜素的方法,然后开展了酸溶辅助超声波法的单因素和正交实验,最后进行了重复性实验。结果表明,酸溶辅助超声波法是较优的提取方法,丙酮是较好的提取溶剂。最佳提取条件为料液比110、超声波处理总时间20 min、酸浓度3 mol/L、酸溶时间25 min、超声波振幅40%、超声工作/间隔时间2 min/1 min、酸溶温度27℃,实验重现较好。优化后类胡萝卜素的提取率较优化前提高了74.8%,为其产业化创造了条件。     

纤溶活性蛋白检测方法研究进展

迄今为止,对纤溶活性蛋白(fibrinolytic protein)的检测主要有4种方法:纤维平板法(fibrin plate method)、显色底物法(colorimetric assay using chromogenic substrates)、反相纤维蛋白自显影法(reverse fibrin autography)和纤维蛋白酶谱法(fibrin zymography)。纤维平板法可以用来快速判断样品是否具有纤溶活性,同时,纤维平板法和显色底物法也可以用来对纤溶活性蛋白进行半定量分析。反相自显影法和纤维蛋白酶谱法则是两种较新的技术,主要用来对纤溶活性蛋白进行定性分析。详细阐述了这几种技术的发展过程、原理、优缺点和应用范围。     

赤爱胜蚯蚓（Eiseniba Foetida Sarigng）纤溶酶的研究──■蚯蚓纤溶酶活性测定方法的研究

本文主要阐述了蚯蚓纤溶酶活性（效价）的测定方法的研究结果。通过对气泡上升法,精氨酸脂酶法（ＴＡＭＥ法）和纤维蛋白平板法测定蚯蚓纤溶酶活性的比较,最后认为纤维蛋白平板法能够客观地表示出纤溶酶溶解血栓的能力,虽然方法较为复杂,难度较大,但准确性强,结果可靠。ＴＡＭＥ法由于受样品中混有杂蛋白酶其稳定性受到一定的影响,但此法简便易行可作为中间产品的快速检测。文中详细地介绍了纤维蛋白平板法。     

一种确定体积氧传递系数kLa的简便方法

通过将稳态法和动态法结合起来,本文提出一种简便的计算Kla的方法,这种方法仅使用稳态法的公式和动态停止通气阶段的数据。由于引入相对溶氧浓度的概念,数据可直接从溶氧电极输出获得。本文方法的最大优点是避免了电极滞后的校正问题。实验结构表明本文方法比需要数值求导的Koizumi的方法更准确。     

SDS-PAGE纤维蛋白自显影法检测不同分子量的纤溶酶原激活剂

体内存在两种纤溶酶原激活剂(plasminogen activator, PA):血液中生理性的组织型纤溶酶原激活剂(t PA)及尿中的尿激酶型纤溶酶原激活剂(u PA)。它们通过将纤溶酶原转变成有活性的纤溶酶而启动纤溶过程使血栓溶解。目前模拟体内纤溶过程以纤维蛋白为底物的PA物质活性测定方法有纤维蛋白溶圈法和发光分析法,但这两种方法不能     

一种改良的小麦醇溶蛋白电泳法

在小麦种子中,醇溶蛋白约占蛋白总量的50％,可分为40—50个组份,每种醇溶蛋白基因已定位于一定染色体上。近年来,世界各国对小麦醇溶蛋白进行电泳分析,发现不同小麦品种之间醇溶蛋白的差异很大,因此醇溶蛋白电泳法可作为小麦品种鉴     

匹拉米洞法、邻联甲苯胺法和联苯胺法 检测林麝便隐血的比较

圈养林麝(Moschusberezovskii)长期受困于消化道类疾病,尤其是肠道炎症性疾病患病率和死亡率一直居高不下。粪便检测是评估野生动物消化系统是否存在出血情况的有效方法之一,并且在圈养林麝肠道健康状况评估及肠道炎症性疾病临床诊断等方面提供了一定的诊断依据。粪便隐血在消化道出血诊断中有广泛的临床诊断价值。基于此,本研究用新鲜的林麝血液进行稀释,来探究匹拉米洞法、邻联甲苯胺法和联苯胺法三种方法对林麝血液浓度的灵敏度范围。检测结果显示,匹拉米洞法的最低敏感性检测浓度为0.05 mg/L,敏感性范围远大于邻联甲苯胺法(0.40 mg/L)和联苯胺法(100.00 mg/L)。分别利用三种检测方法对林麝粪便潜血进行检测,比较检测结果的阳性率,结果显示,匹拉米洞法的检测效果优于其他两种方法,阳性率分别为匹拉米洞法检测法10.13%、邻联甲苯胺法检测法2.56%和联苯胺法检测法0,差异有诊断学意义(P 0.05)。而且在操作上,匹拉米洞法更加简便快捷。故在诊断林麝消化道出血时,采用匹拉米洞法进行林麝便隐血的检测更加准确便捷。     

抗肿瘤药物筛选中MTT法和SRB法的比较

在抗肿瘤药物的体外筛选中 ,MTT法和 SRB法是常用的两种方法。我们用MTT法和 SRB法分别测定 3种已知植物抗癌药对 2 2株人肿瘤细胞的抗癌活性 ,对这两种方法进行了详细的比较。通过分析两种方法测出的细胞存活率 ( T/ C)的差异分布和相关系数以及 IC50 的二变量分布 ,比较了两种方法测定结果的异同 ;通过两种方法重复测定 3种药物对 7株人癌细胞的抗癌活性 ,比较了两种方法的重复性 ;通过分析两种方法测定结果 T/ C值随时间变化的程度 ,比较了两种方法测定结果的稳定性。实验结果表明 :MTT法和 SRB法的相关性较好 ,都可用于抗肿瘤药物的体外筛选 ,SRB法更适合于大规模筛选 ,3种抗癌药物的测定结果与临床资料基本一致。     

条件频率法的改进──加权频率法

条件频率法和综合相关法[1,2]都是将预报因子对预报对象的作用等同对待。在许多实际问题中,各预报因子所起的作用往往是不同的,用条件频率法或综合相关法来预测,无疑会影响其预报的准确性。为了解决这一问题,作者在本文中提出了一种新的简便预测方法──加权频率法。1基本原理与方法首先将原始资料依据分级标准进行分级。设有m个预报因子工人xj（j=1,2,……,m）分为s级,预报对象y分为r级。然后根据各预报因子与预报对象分级数码制作额数列联表,将xj处于k（k=1,2,……,s）级时的总个数,以nk·表示;将xj处于k级时,y=1（1=1,2…     

洗胃法与剖胃法在四种蛙食性分析中的对比

在动物的食性研究中,剖胃法因要杀死动物,会产生一系列的生态学影响和伦理学问题,在国际上招致了越来越多的反对,而在我国仍广泛使用。尽管洗胃法已用于食性研究,它的效果却从来没有研究过。我们应用洗胃法研究了浙江宁波郭巨镇四种蛙(黑斑蛙、泽蛙、金线蛙和中华蟾蜍)的食性,并与剖胃法所获得的食性结果进行比较。结果显示:尽管洗胃法在四种蛙胃内食物量的总洗出率(在94.50%以上)和食物个体数的洗出率上(均大于93.46%)略低于剖胃法,但胃内未洗出的与洗出的平均食物个体大小间无差异(所有P>0.3061),两种方法获得的四种蛙的平均食物量和食物个体数也无差异(对食物量所有P>0.8680,对食物个体数所有P>0.7923)。结果表明,洗胃法是比较精确的,在动物的食性研究中,应广泛地推广洗胃法,摒弃剖胃法     

Development and Validation of a Discriminating <Emphasis Type="Italic">In Vitro</Emphasis> Dissolution Method for a Poorly Soluble Drug,Olmesartan Medoxomil: Comparison Between Commercial Tablets

A dissolution test for tablets containing 40 mg of olmesartan medoxomil (OLM) was developed and validated using both LC-UV and UV methods. After evaluation of the sink condition, dissolution medium, and stability of the drug, the method was validated using USP apparatus 2, 50 rpm rotation speed, and 900 ml of deaerated H₂O + 0.5% sodium lauryl sulfate (w/v) at pH 6.8 (adjusted with 18% phosphoric acid) as the dissolution medium. The model-independent method using difference factor (f ₁) and similarity factor (f ₂), model-dependent method, and dissolution efficiency were employed to compare dissolution profiles. The kinetic parameters of drug release were also investigated. The obtained results provided adequate dissolution profiles. The developed dissolution test was validated according to international guidelines. Since there is no monograph for this drug in tablets, the dissolution method presented here can be used as a quality control test for OLM in this dosage form, especially in a batch to batch evaluation.     

Development of a New Dissolution Test Method for Soft Chewable Dosage Forms

Soft chewable dosage forms are a new approach to improve the compliance of medication for special patient populations. Based on their texture, they are chewed several times before they get swallowed. A suitable dissolution method based on in vivo chewing data was developed. The method covers parts of dissolution within the oral cavity (simulation of chewing) as well as the dissolution of the swallowed bolus within the gastrointestinal tract. Chewing was simulated by the help of a steel tooth assembled to a texture analyzer and wedge gliding on an inclined plane, imitating the occlusal glide. Chewing cycles of non-brittle, elastically deformable foods were predicted by a multiple linear regression (R _adj?=?0.985) using hardness, stickiness, fat/water content, and softening behavior as independent variables. Cross-validation of three sets of chewing data led to a root mean square error of prediction of 0.408 or 0.658 chewing cycles, respectively. The new method is able to distinguish between different soft chewable formulations which had been approved as similar by the dissolution method of the European Pharmacopoeia. Furthermore, it provides information about the drug content released within the time of chewing (7–15%).     

Development and Validation of a HPLC Method for Dissolution and Stability Assay of Liquid-Filled Cyclosporine Capsule Drug Products

To assay the dissolution samples of a drug product from several sources, a simple but broadly applicable analytical method is always desired. For the liquid-filled cyclosporine capsules, while analyzing the dissolution samples, the current compendial and literature HPLC methods have been found to be inadequate to provide satisfactory separation of the drug and the excipient peaks. Accordingly, a suitable isocratic reverse-phase HPLC method was developed for the analysis of dissolution samples of liquid-filled cyclosporine capsules. The method successfully separated the cyclosporine peak from the interfering chromatographic peaks of the excipients. The method was validated according to the ICH and FDA guidelines. Specificity, selectivity, linearity, accuracy, precision, and robustness were established over 3 days as part of method validation. Additionally, the degradation kinetics of cyclosporine in dissolution media was determined. Cyclosporine degradation followed a zero-order kinetics in the dissolution media with the respective rate constants of −3.5, −1.5, and −0.3%/h at 37°C, 25°C, and 10°C.Key words: cyclosporine, degradation, dissolution, HPLC, liquid-filled capsules     

An Investigation into the Influence of Experimental Conditions on In Vitro Drug Release from Immediate-Release Tablets of Levothyroxine Sodium and Its Relation to Oral Bioavailability

The aim of this study was to investigate the influence of experimental conditions on levothyroxine sodium release from two immediate-release tablet formulations which narrowly passed the standard requirements for bioequivalence studies. The in vivo study was conducted as randomised, single-dose, two-way cross-over pharmacokinetic study in 24 healthy subjects. The in vitro study was performed using various dissolution media, and obtained dissolution profiles were compared using the similarity factor value. Drug solubility in different media was also determined. The in vivo results showed narrowly passing bioequivalence. Considering that levothyroxine sodium is classified as Class III drug according to the Biopharmaceutics Classification System, drug bioavailability will be less sensitive to the variation in its dissolution characteristics and it can be assumed that the differences observed in vitro in some of investigated media probably do not have significant influence on the absorption process, as long as rapid and complete dissolution exists. The study results indicate that the current regulatory criteria for the value of similarity factor in comparative dissolution testing, as well as request for very rapid dissolution (more than 85% of drug dissolved in 15 min), are very restricted for immediate-release dosage forms containing highly soluble drug substance and need further investigation. The obtained results also add to the existing debate on the appropriateness of the current bioequivalence standards for levothyroxine sodium products.KEY WORDS: bioequivalence, dissolution, immediate release, levothyroxine sodium, solubility     

An Investigation into the Influence of Experimental Conditions on <Emphasis Type="Italic">In Vitro</Emphasis> Drug Release from Immediate-Release Tablets of Levothyroxine Sodium and Its Relation to Oral Bioavailability

The aim of this study was to investigate the influence of experimental conditions on levothyroxine sodium release from two immediate-release tablet formulations which narrowly passed the standard requirements for bioequivalence studies. The in vivo study was conducted as randomised, single-dose, two-way cross-over pharmacokinetic study in 24 healthy subjects. The in vitro study was performed using various dissolution media, and obtained dissolution profiles were compared using the similarity factor value. Drug solubility in different media was also determined. The in vivo results showed narrowly passing bioequivalence. Considering that levothyroxine sodium is classified as Class III drug according to the Biopharmaceutics Classification System, drug bioavailability will be less sensitive to the variation in its dissolution characteristics and it can be assumed that the differences observed in vitro in some of investigated media probably do not have significant influence on the absorption process, as long as rapid and complete dissolution exists. The study results indicate that the current regulatory criteria for the value of similarity factor in comparative dissolution testing, as well as request for very rapid dissolution (more than 85% of drug dissolved in 15 min), are very restricted for immediate-release dosage forms containing highly soluble drug substance and need further investigation. The obtained results also add to the existing debate on the appropriateness of the current bioequivalence standards for levothyroxine sodium products.     

Development and Validation of a Discriminative Dissolution Method for Atorvastatin Calcium Tablets using in vivo Data by LC and UV Methods

A dissolution method to analyze atorvastatin tablets using in vivo data for RP and test pilot (PB) was developed and validated. The appropriate conditions were determined after solubility tests using different media, and sink conditions were established. The conditions used were equipment paddle at 50 rpm and 900 mL of potassium phosphate buffer pH 6.0 as dissolution medium. In vivo release profiles were obtained from the bioequivalence study of RP and the generic candidate PB. The fraction of dose absorbed was calculated using the Loo–Riegelman method. It was necessary to use a scale factor of time similar to 6.0, to associate the values of absorbed fraction and dissolved fraction, obtaining an in vivo–in vitro correlation level A. The dissolution method to quantify the amount of drug dissolved was validated using high-performance liquid chromatography and ultraviolet spectrophotometry, and validated according to the USP protocol. The discriminative power of dissolution conditions was assessed using two different pilot batches of atorvastatin tablets (PA and PB) and RP. The dissolution test was validated and may be used as a discriminating method in quality control and in the development of the new formulations.     

Application of Absorption Modeling to Predict Bioequivalence Outcome of Two Batches of Etoricoxib Tablets

As part of the overall product development and manufacturing strategy, pharmaceutical companies routinely change formulation and manufacturing site. Depending on the type and level of change and the BCS class of the molecule, dissolution data and/or bioequivalence (BE) may be needed to support the change for immediate release dosage forms. In this report, we demonstrate that for certain weakly basic low-solubility molecules which rapidly dissolve in the stomach, absorption modeling could be used to justify a BE study waiver even when there is failure to show dissolution similarity under some conditions. The development of an absorption model for etoricoxib is described here, which was then used to a priori predict the BE outcome of tablet batches manufactured at two sites. Dissolution studies in 0.01 N HCl media (pH 2.0) had demonstrated similarity of etoricoxib tablets manufactured at two different sites. However, dissolution testing at pH 4.5 and pH 6.8 media failed to show comparability of the tablets manufactured at the two sites. Single simulations and virtual trials conducted using the 0.01 N HCl dissolution showed similarity in AUC and C_max for all tablet strengths for batches manufactured at the two manufacturing sites. These predicted results were verified in a definitive bioequivalence study, which showed that both tablet batches were bioequivalent. Since the development of traditional in vitro–in vivo correlations (IVIVC) for immediate release (IR) products is challenging, in cases such as etoricoxib, absorption modeling could be used as an alternative to support waiver of a BE study.KEY WORDS: bioequivalence, dissolution, modeling, pharmacokinetics, SUPAC     

Gastroretentive drug delivery system of ranitidine hydrochloride: Formulation and in vitro evaluation

The purpose of this research was to prepare a gastroretentive drug delivery system of ranitidine hydrochloride. Guar gum, xanthan gum, and hydroxypropyl methylcellulose were evaluated for gel-forming properties. Sodium bicarbonate was incorporated as a gas-generating agent. The effects of citric acid and stearic acid on drug release profile and floating properties were investigated. The addition of stearic acid reduces the drug dissolution due to its hydrophobic nature. A 3² full factorial design was applied to systemically optimize the drug release profile. The amounts of citric acid anhydrous (X₁) and stearic acid (X₂) were selected as independent variables. The times required for 50% (t₅₀) and 80% drug dissolution (t₈₀), and the similarity factor f₂ were selected as dependent variables. The results of the full factorial design indicated that a low amount of citric acid and a high amount of stearic acid favors sustained release of ranitidine hydrochloride from a gastroretentive formulation. A theoretical dissolution profile was generated using pharmacokinetic parameters of ranitidine hydrochloride. The similarity factor f₂ was applied between the factorial design batches and the theoretical dissolution profile. No significant difference was observed between the desired release profile and batches F2, F3, F6, and F9. Batch F9 showed the highest f2 (f2=75) among all the batches, and this similarity is also reflected in t₅₀ (∼214 minutes) and t₈₀ (∼537 minutes) values. These studies indicate that the proper balance between a release rate enhancer and a release rate retardant can produce a drug dissolution profile similar to a theoretical dissolution profile.     

SCGPred： A Score-based Method for Gene Structure Prediction by Combining Multiple Sources of Evidence

Predicting protein-coding genes still remains a significant challenge. Although a variety of computational programs that use commonly machine learning methods have emerged, the accuracy of predictions remains a low level when implementing in large genomic sequences. Moreover, computational gene finding in newly se- quenced genomes is especially a difficult task due to the absence of a training set of abundant validated genes. Here we present a new gene-finding program, SCGPred, to improve the accuracy of prediction by combining multiple sources of evidence. SCGPred can perform both supervised method in previously well-studied genomes and unsupervised one in novel genomes. By testing with datasets composed of large DNA sequences from human and a novel genome of Ustilago maydi, SCGPred gains a significant improvement in comparison to the popular ab initio gene predictors. We also demonstrate that SCGPred can significantly improve prediction in novel genomes by combining several foreign gene finders with similarity alignments, which is superior to other unsupervised methods. Therefore, SCGPred can serve as an alternative gene-finding tool for newly sequenced eukaryotic genomes. The program is freely available at http：//bio.scu.edu.cn/SCGPred/.     

Dissolution Testing of Sublingual Tablets: A Novel <Emphasis Type="Italic">In Vitro</Emphasis> Method

In the sublingual (SL) cavity, compared with the gastrointestinal tract, tablets are subjected to minimal physiological agitation, and a limited volume of saliva is available to facilitate disintegration and dissolution. None of the official compendial dissolution apparatuses and methods simulate these SL conditions. In this study, a custom-made dissolution apparatus was constructed, and a novel in vitro method that simulates SL conditions was evaluated. Several epinephrine 40 mg SL tablet formulations under development and two commercial SL tablets, isosorbide dinitrate 5 mg and nitroglycerin 0.6 mg, were studied. The dissolution medium was 2 mL of distilled water at 25°C. Dissolution was measured at 60 and 120 s. The novel in vitro method was validated for accuracy, reproducibility, and discrimination capability, and was compared with the official US Pharmacopeia (USP) dissolution method using apparatus 2 (Paddle). The data obtained following the novel in vitro method were accurate and reproducible. This method was capable of detecting minor changes in SL formulations that could not be detected using other in vitro tests. Results from the official USP dissolution method and our novel in vitro method were significantly different (p < 0.05). Results reflecting the dissolution of rapidly disintegrating tablets using simulated SL conditions were obtained using the novel in vitro dissolution method.