不同方法提取的佩兰挥发油指纹图谱分析

目的:利用GC-MS分析了三种方法提取的佩兰挥发油指纹图谱。方法:采取超临界CO2萃取、微波萃取和水蒸汽蒸馏三种方法提取佩兰挥发油。结果:三种方法提取佩兰挥发油的化学成分均有差异。结论:超临界CO2萃取的佩兰挥发油更具天然性,超临界CO2萃取法为提取佩兰挥发油的较理想方法;微波辅助萃取也不失为一种可行的方法。     

零捻纱和变形纱的上浆

将连续长丝加工成可织造纱线有三种方法:混合、加捻和上浆。与第三种方法相比,前两种方法比较便宜,但这两种方法改变了长丝的表面特性,而第三种方法(即上浆)则不会改变长丝的表面特性,却能改善其织造性能。对于需要挺爽手感的织     

几种HPLC法测定银杏总黄酮的比较研究

使用HPLC法测定了银杏黄酮总量,研究了3种检测方法.方法一是以槲皮素、山柰酚、异鼠李素三种苷元为标样进行HPLC检测,通过计算3种苷元的总和得到总黄酮含量;方法二是以槲皮素为标样进行HPLC检测,采用归一化法计算黄酮总量;方法三是以槲皮素为标样进行HPLC检测,利用总黄酮与槲皮素含量的系数来计算总黄酮含量的方法.通过对三种HPLC检测方法的研究,比较了各自优缺点,确定各自适用范围.     

甜菜施用稀土技术试验简报

试验证明:拌种、浸种、叶喷三种施用稀土的方法都可以使甜菜的块根产量、含糖率增加。三种方法平均亩增广8.8%,增糖1.0度。增加产糖量15.5%。平均亩增加收入50.33元。三种施用方法以稀土拌种效果最好,施用剂量以亩拌40克稀土为宜。     

三种检测啤酒中二氧化硫的方法比较

目前有几种比较成熟的检测啤酒中二氧化硫的方法，如：国标第一法即氧化法，国标第二法即碘量法以及本文中的方法三。本实验的目的是通过三种方法的回收率和重现性比较实验，分析三种方法的优缺点，从而确定实验室检测啤酒中二氧化硫的最佳方法。     

不同方法定量检测花椒油中花椒麻味物质的效果比较研究

本文应用气相色谱法、高效液相色谱法、分光光度计法对花椒麻味物质的含量进行了定量检测分析,并比较了三种检测方法的技术指标以及在实际检测中的效果。结果发现三种检测方法在实际应用中各有其自身的优缺点,从技术指标来看,气相色谱法、高效液相色谱法要优于分光光度计法,在实际的样品检测中,前两种方法也要比分光光度计法更加准确,但相比较而言分光光度计法的使用成本要低于前两种检测方法。通过对三种方法的差异显著性分析发现,在检测不同样品时,三种方法的差异显著性也存在不同。在实际运用中应该从对结果的准确性与实际经济性的比较中进行考虑选择合理的检测方法。     

糖基化方法对于大豆7S球蛋白糖基化产物构象及功能特性的影响

针对传统的两种糖基化接枝方法和一种改进方法从糖基化产物构象和产物功能特性方面对三种糖基化方法进行研究比较,以期进一步揭示不同糖基化方法对于蛋白质糖基化反应及产物的影响机制,结果表明:水相体系比固相体系对于蛋白质空间结构影响更大,二级结构中,湿热法和水热法的制备产物α-螺旋含量均减少,三种糖基化产物二级结构的主要结构与大豆7S球蛋白一样仍以β-结构为主,接入糖链导致蛋白质的二级结构无规则卷曲含量都增加;三级结构中,干热法对于三级结构的变化影响不明显,然而湿热法和水热法对于蛋白质分子的三级结构有显著的影响;三种糖基化产物的主要功能特性溶解性、乳化性和热稳定性均有提高,干热法产物在溶解性和乳化性方面均优于其它两种方法,热稳定性三种方法差异性不大。     

运用ASME法进行法兰泄漏分析

目前,被广泛使用的较为成熟的法兰泄漏分析方法主要有三种:当量压力法、NC3658.3法和ASME B&PVC规范Ⅷ-1卷附录2法.前两种方法主要适用于一般工况下法兰分析,对于特殊高温高压或低温工况以及非标法兰一般采用第三种方法.文章通过分析法兰密封的原理,从根本上找到影响泄漏的主要因素,并主要分析第三种方法的基本原理,...     

三种检测酒精度方法的比较

研究了蒸馏法、Anton Paar、SCABA三种方法检测酒精度的原理，分析了三种方法的检测数据，确定了SCABA是酒精度检测的最佳方法。     

染整工艺碱液的测控

论述了工艺碱液的折射法、电导法及比重法测量机理;阐明了三种方法的传感器结构;介绍了三种方法的自动控制系统及多组份化学助剂的计量配液。     

16S rRNA基因序列、生化鉴定、质谱3类方法鉴定常见微生物的结果分析

目的比较16SrRNA基因序列、生化鉴定、质谱鉴定3类实验方法分析沙门氏菌、金黄色葡萄球菌、蜡样芽孢杆菌的鉴定结果的异同。方法挑选沙门氏菌、金黄色葡萄球菌、蜡样芽孢杆菌各50株,分别进行16S rRNA基因序列测定、VITEK COMPACT 2生化鉴定、质谱鉴定3类实验,并比较3类实验的鉴定结果。结果 3类实验方法对大部分沙门氏菌鉴定在属水平,生化鉴定方法对少数血清型鉴定到种水平;对金黄色葡萄球菌均鉴定到种水平; 16SrRNA基因序列、生化方法对蜡样芽孢杆菌鉴定到属水平,质谱鉴定到种水平。结论 3类实验方法对大部分沙门氏菌、所有金黄色葡萄球菌鉴定水平相同,质谱鉴定蜡样芽孢杆菌的结果更准确,且质谱鉴定时效性更高。     

克罗诺杆菌属食品分离株种水平鉴定方法比较研究

目的研究食品中分离克罗诺杆菌属种的鉴定方法,旨在建立稳定、可靠的种的鉴定方法。方法本研究选择三种鉴定方法,包括生化鉴定(VITEK 2 Compact)法、基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)、全基因组测序,通过比较分析三种方法对19株克罗诺杆菌属的鉴定结果,阐述三种鉴定方法的优缺点及其适用性。结果本研究中19株克罗诺杆菌属的鉴定结果显示,VITEK 2 Compact对阪崎克罗诺杆菌和丙二酸盐克罗诺杆菌的鉴定准确率只有67%和66%,而MALDI-TOF MS和全基因组测序的鉴定结果一致,因此可以判定MALDI-TOF MS能够快速、准确、高通量对克罗诺杆菌属进行种的鉴定。但是MALDI-TOF MS受限于数据库,不能鉴定到亚种的水平。结论本研究结果提示VITEK 2 Compact法不适用于克罗诺杆菌属种的鉴定;全基因组测序鉴定结果准确可靠;MALDI-TOF MS可以实现快速、高通量、准确的鉴定,仍需要进一步研究克罗诺杆菌属3个亚种的蛋白指纹图谱特点,丰富蛋白指纹图谱数据库,使其能够准确鉴定克罗诺杆菌属的7个种和3个亚种。     

金黄色葡萄球菌基质辅助激光解析电离-飞行时间质谱的鉴定与聚类分型

目的 建立基质辅助激光解吸电离-飞行时间质谱法(matrix assisted laser desorption lonization-time of flight mass spectrometer, MALDI-TOF MS)快速鉴定金黄色葡萄球菌(Staphylococcus aureus, S. aureus)并自建数据库, 进行聚类分型。方法 基于MALDI-TOF MS技术, 对经全自动微生物分析仪(VITEK2 COMPACT)鉴定为S. aureus的25株分离株和1株标准菌株(ATCC 25923)进行鉴定。并进一步采集26株S. aureus特征性蛋白质指纹图谱, 使用flex Analysis软件进行分析, 汇总成标准图谱并构建本地分离株S. aureus的鉴定数据库, 命名为Staphylococcus aureus。结果 经标准菌株ATCC 25923验证, 自建数据库对S. aureus鉴定结果的可信度较设备自带数据库明显提高, 鉴定分值由2.251提高到了2.845。自建数据库进一步对26株S. aureus进行聚类分型, 在差异水平100时, 24株不同来源S. aureus被聚类成24个分支, 将食品中不同来源分离的S. aureus分为24个型。结论 MALDI-TOF MS作为一种快速、准确、高通量的全新微生物鉴定技术, 实现了对S. aureus的特异性快速鉴定与分型, 能够满足公共安全卫生、突发食品安全事件和口岸快速通关方面的需求。     

预包装食品中金黄色葡萄球菌3种鉴定方法的比较

目的 比较国家标准方法(GB 4789.10-2016)、VITEK2 Compact全自动微生物鉴定系统、基质辅助激光解吸电离-飞行时间质谱法对预包装食品中金黄色葡萄球菌的鉴定效果。方法 使用金黄色葡萄球菌(ATCC 25923)人工污染12种不同种类的预包装食品,按照GB4789.10-2016进行样品前处理和增菌,通过上述三种方法对上述样品中分离的疑似金黄色葡萄球菌进行鉴定,并对鉴定结果进行比较。结果 三种方法均能够对12种不同种类的人工污染预包装食品中分离的金黄色葡萄球菌进行准确有效的鉴定。相对于国标法,两种仪器鉴定方法无需进行后续溶血分析和血浆凝固酶试验,对操作人员技术要求更低,能够在更短的时间内获得鉴定结果。结论 两种仪器鉴定方法快速、可靠且操作简单,可以作为国标法的有效补充。     

Matrix-assisted laser desorption and ionization-time-of-flight mass spectrometry, 16S rRNA gene sequencing, and API 32E for identification of Cronobacter spp.: a comparative study

Twenty-two isolates of the family Enterobacteriaceae, with focus on Cronobacter isolated from infant formula and the environment of milk powder plants, were comparatively identified using API 32E (bioMérieux, Marcy l'Etoile, France), 16S rRNA gene sequencing (Accugenix, Newark, USA), and matrix-assisted laser desorption and ionization-time-of-flight mass spectrometry (MALDI-TOF MS; Mabritec, Riehen, Switzerland and AnagnosTec, Potsdam, Germany). With API 32E, 22% of the isolates were assigned to species, 64% were assigned to a genus, and 14% could not be discriminated at any taxonomic level. Both 16S rRNA gene sequencing and MALDI-TOF MS assigned 100% of the isolates to species, but the identifications based on MALDI-TOF MS results were more discriminating and unequivocal. Our data indicate that MALDI-TOF MS provides the most rapid and unambiguous identification of Cronobacter and closely related Enterobacteriaceae isolates.     

Bovine mastitis bacteria resolved by MALDI-TOF mass spectrometry

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a fast and reliable method to identify the most common pathogenic bacteria in humans and animals. The goals of this study were to amend a commercial database with additional species, evaluate the amended database for identification of bacterial genera and species causing bovine mastitis, and describe the plethora of species involved. In total, 500 udder pathogenic isolates were subjected to MALDI-TOF MS using bacterial or fungal colony material; 93.5% could be identified to the species level, and 6.5% were identified only to the genus level. Isolates identified to the genus level required further identification to the species level by conventional methods or 16S rDNA sequencing. Mass spectra from verified species were used to expand the MALDI-TOF MS database to improve future identification ability. A total of 24 genera and 61 species were identified in this study. Identified isolates were mainly staphylococci, streptococci, Enterobacteriaceae, and coryneforme bacteria. In conclusion, MALDI-TOF MS is a powerful, rapid, and reliable technique to identify the most common microorganisms causing bovine mastitis, and the database can be continuously expanded and improved with additional species.     

Differentiation of Staphylococci from Iberian dry fermented sausages by protein fingerprinting

The Staphylococci populations in different types of Iberian dry fermented sausages from central-west Spain were identified. A simple electrophoretic method of whole-cell proteins and extracellular protein profiling was evaluated for speed of identification. This study was correlated with a 16S rRNA gene sequence analysis and biochemical identification by API Staph. A total of 81 isolates were identified by SDS-PAGE of the whole-cell proteins. These showed stable profiles in the range 99-14kDa that were clearly different for the different species, and were grouped into clusters together with the profiles of the eight reference strains. SDS-PAGE of the extracellular protein extracts provided additional characteristic banding patterns for the characterization of the Staphylococcus species present. The whole-cell SDS-PAGE showed that the predominant species was Staphylococcus saprophyticus (61.7%) followed by Staphylococcus aureus (19.7%). The identifications were confirmed by the 16S rRNA gene sequencing and by a BLAST search of the GenBank database. However, the API Staph biochemical identifications were frequently erroneous at the species level. In sum, SDS-PAGE analysis showed itself to be rapid and accurate in identifying the most commonly encountered Staphylococcus isolates in dry fermented sausages.     

金黄色葡萄球菌鉴定方法研究

目的 为了筛选出能够快速准确鉴定金黄色葡萄球菌的方法。方法 利用国标法、VITEK 2 Compact生化鉴定、16s rDNA序列分析和PCR鉴定等4种方法对酱卤肉制品中分离出的典型菌落进行鉴定。结果 国标法检测结果显示菌株为革兰氏阳性菌,血平板上有明显的透明溶血圈且血浆凝固酶试验结果为阳性,符合金黄色葡萄球菌判定标准。VITEK 2 Compact生化鉴定结果中不吻合的典型生化谱仅有1项（dMAL）,判定菌株为金黄色葡萄球菌（98%概率）,为极好的鉴定。16s rDNA序列比对分析及以邻接法构建的进化树均显示典型菌落为金黄色葡萄球菌。以耐热核酸酶基因（nuc）设计的引物能扩增出单一的清晰条带,能快速准确的识别出金黄色葡萄球菌。结论 4种方法均能准确鉴定出金黄色葡萄球菌,但耗时都比较长,通过改进实验方法,缩短DNA提取时间,PCR鉴定将表现出较大优势。     

鉴定乳制品中厌氧芽胞杆菌的3种方法比较

鉴定乳制品中的革兰氏阳性厌氧芽胞杆菌。方法 采用生化鉴定、基质辅助激光解析电离飞行时间质谱(MALDI-TOF MS)鉴定和16S rDNA测序三种方法,对乳制品中5株革兰氏阳性厌氧芽胞杆菌进行鉴定。结果 生化方法鉴定出3株菌,而飞行时间质谱鉴定和16S rDNA测序法对5株菌都给出了鉴定结果。3种方法仅对其中1株菌的鉴定结果一致;飞行时间质谱鉴定和16S rDNA测序的鉴定结果一致性程度高,共有4株菌的鉴定结果一致;而生化方法的鉴定结果与这两种方法差异较大。结论 飞行时间质谱鉴定和16S rDNA测序可作为生化方法的补充,用于革兰氏阳性厌氧芽胞杆菌的快速鉴定。     

Rapid species- and subspecies-specific level classification and identification of Lactobacillus casei group members using MALDI Biotyper combined with ClinProTools

Using common taxonomic methods such as 16S rDNA sequencing and physiological and biochemical analysis to identify members of the Lactobacillus casei group (LCG) is time-consuming, expensive and inaccurate. In this study, we applied matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to rapid discriminate LCG strains by creating an analytical in-house database (IHDB) and to develop a classification model for subspecies-level differentiation based on MS biomarkers using ClinProTools bioinformatics software (Bruker Daltonics, Billerica, MA). Genotypic methods (housekeeping gene sequencing and species-specific PCR) were also established to validate the MALDI-TOF MS platform. A total of 48 LCG reference strains were correctly identified at the species level (mean score: 2.45 ± 0.1) by using MALDI-TOF MS with an IHDB and had high score values, which was in accordance with results from mutL gene sequencing and specific PCR-based methods. However, one strain that was identified as L. casei had a relatively low score value (2.02 ± 0.02), lower sequence similarities (mutL: 90.4%), and failed to amplify a species-specific amplicon; it may therefore represent an undescribed novel species. In addition, after implementation of the classification model (based on 2 biomarker peaks: m/z 4,930 and 5,303), L. paracasei strains could be clearly and easily differentiated to the subspecies level. Afterward, 7 LCG-related isolates from different probiotic samples were analyzed and accurately identified. Our data demonstrate the high-resolution performance of MALDI-TOF MS for fast and accurate demarcation of LCG strains when used with an IHDB coupled to ClinProTools; this methodology can serve as an alternative for quality control of probiotic products.