Absolute Spectrophotometry of Northern Compact Planetary Nebulae
高效液相色谱-二级管阵列检测器法测定美白类、祛斑类化妆品和香波中氢醌、苯酚含量
摘要
样 品用甲醇超声提 取 1 i, 5mn 离心、 滤, A in O B X E l s D 过 以 ge t R A ci eX B—C l Z p N分析柱分 离分析 , 二级 管
阵列检测 器检 测 , 外标 法定量 。氢醌、 苯酚分别在 7 9 0— 9 90mg L 9 6 6—9 .6 q/ . 2 7 .2 / 、 .9 6 9 0n L浓度 范围 内线 性关 系 I g
1 实验 部分
1 1 主 要 仪 器 与 试 剂 .
高效 液 相色 谱仪 : get 2 0型 , A in 10 l 附带 D D二 A 极 管 阵列检 测器 , 国安捷 伦公 司 ; 美
超 声波 清洗 器 : 7 2 4型 , 国 Em D一 8 2 德 l a公 司 ; 高速离 心机 : 3 K型 , 国 Sg a 司 ; 3— 0 德 im 公 分 析天 平 : L 0 A 2 4型 , 特 勒 一托 利 多仪 器 ( 梅 上 海 ) 限公 司 ; 有
测 检 验祛 斑美 白类 化 妆 品 的质 量 , 证 消 费 者用 到 保
分别精密称取氢醌 、 苯酚标准品 0 09 0 1 1 .9 、.22 g用 甲醇溶解后定容至 10m , 0 L棕色容量瓶中, 分别
得 到 0 90、. 1 g m .9 12 2m / L的 氢 醌 、 酚 标 准 储 备 苯 液 , 4C暗处 保 存 。临 用 时 将 标 准 储 备 液 用 甲 醇 于  ̄ 稀 释成所 需 浓度 的混合 标准 工作 溶液 。
2 2 提 取 溶 剂 .
利 用 氢醌 和苯 酚标准 储备 液配 制成 氢醌 浓度依
次 为 7 9 0 1 . 4 、 1 6 0 3 . 0 、9 2 0 mg L . 2 、5 8 0 3 . 8 、9 6 0 7 . 0 / , 苯 酚 浓 度 依 次 为 9 6 6 1 .9 、8 74 4 . 8 、 . 9 、 9 3 2 3 . 8 、 8 4 0
211133774_表面增强拉曼光谱法快速检测保健食品中非法添加药物盐酸氨溴索
表面增强拉曼光谱法快速检测保健食品中非法添加药物盐酸氨溴索刘 虎1,刘 猛1,刘 杰1,郑博士1,常化仿2,程金新1*(1.中国人民警察大学 侦查学院,河北廊坊 065000;2.普拉瑞思科学仪器(苏州)有限公司,江苏苏州 215004)摘 要:本文采用表面增强拉曼技术检测保健品中非法添加的盐酸氨溴索。
结果表明,峰值强度具有良好线性关系,检测限为0.2 mg·L-1,定量限范围为0.5~5.0 mg·L-1。
该方法快速简便,适用于止咳平喘类保健品中盐酸氨溴索的定性定量分析。
关键词:盐酸氨溴素;保健品;表面增强拉曼光谱Rapid Detection of Illegal Addition of Ambroxol Hydrochloride by Surface-Enhanced Raman Spectroscopy in Health Food LIU Hu1, LIU Meng1, LIU Jie1, ZHENG Boshi1, CHANG Huafang2, CHENG Jinxin1*(1.Investigation College of the People’s Police University of China, Langfang 065000, China; 2.Praxis ScientificInstruments (Suzhou) Co., Ltd., Suzhou 215004, China)Abstract: In this paper, surface-enhanced Raman technique was used to detect ambroxol hydrochloride illegally added in health products. The results showed that the peak intensity had a good linear relationship, the detection limit was 0.2 mg·L-1, and the limit of quantitation was 0.5~5.0 mg·L-1. The method is rapid, simple and suitable for qualitative and quantitative analysis of ambroxol hydrochloride in cough and asthma relief health products.Keywords: ambromine hydrochloride; health care products; surface-enhanced Raman spectrum近年来,在保健食品中添加化学药物的违法行为屡禁不止。
2010药典 拉曼光谱法指导原则
拉曼光谱法指导原则拉曼光谱法是研究化合物分子受光照射后所产生的散射,散射光与入射光能级差和化合物振动频率、转动频率的关系的分析方法。
与红外光谱类似,拉曼光谱是一种振动光谱技术。
所不同的是,前者与分子振动时偶极矩变化相关,而拉曼效应则是分子极化率改变的结果,被测量的是非弹性的散射辐射。
拉曼光谱通常采用激光作为单色光源,将样品分子激发到某一虚态,随后受激分子弛豫跃迁到一个与基态不同的振动能级,此时,散射辐射的频率将与入射频率不同。
这种频率变化与基态和终态的振动能级差相当。
这种“非弹性散射”光就称之为拉曼散射。
频率不变的散射称为弹性散射,即所谓瑞利散射。
如果产生的拉曼散射频率低于入射频率,则称之为斯托克散射。
反之,则称之为反斯托克散射。
实际上,几乎所有的拉曼分析都是测量斯托克散射。
拉曼光谱与红外吸收光谱相似。
用散射强度对拉曼位移作图。
拉曼位移(以cm-1 为单位)等于激发光的波数减去散射辐射的波数。
由于功能团或化学键的拉曼位移与它们在红外光谱中的吸收波数相一致,所以谱图的解析也与红外吸收光谱相同。
然而,通常在拉曼光谱中出现的强谱带在红外光谱中却成为弱谱带甚至不出现,反之亦然。
所以,这两种光谱技术常互为补充。
拉曼光谱的优点在于它的快速,准确,测量时通常不破坏样品(固体,半固体,液体或气体),样品制备简单甚至不需样品制备。
谱带信号通常处在可见或近红外光范围,可以有效地和光纤联用。
这也意味着谱带信号可以从包封在任何对激光透明的介质,如玻璃,塑料内,或将样品溶于水中获得。
现代拉曼光谱仪使用简单,分析速度快(几秒到几分钟),性能可靠。
因此,拉曼光谱与其他分析技术联用比其他光谱联用技术从某种意义上说更加简便(可以使用单变量和多变量方法以及校准)。
除常规的拉曼光谱外,还有一些较为特殊的拉曼技术。
它们是共振拉曼,表面增强拉曼光谱,拉曼旋光,相关-反斯托克拉曼光谱,拉曼增益或减失光谱以及超拉曼光谱等。
其中,在药物分析应用相对较多的是共振拉曼和表面增强拉曼光谱法。
Absorption Spectrophotometry
How is absorption spectrophotometry used to measure protein concentration?
Protein concentration is typically measured by combining a small sample of the homogenate with a chemical reagent that changes color in proportion to the amount of protein present. Several commercially prepared protein assay reagents are available, and for this lab we will use the BCA reagent. In the presence of protein turns a purple color – the more protein present the deeper the color. The purplecolored product absorbs light best at 562 nM, the wavelength at which the measurements are made. Thus, 562 nM is the 8max (wavelength of maximum absorbance) for the BCA reagent in the presence of protein? The protein content of the liver homogenate will be determined by comparing the absorbance of homogenate sample to the absorption coefficient determined from a standard curve. A Standard Curve is prepared by plotting the absorbance of samples containing known concentrations of the standard protein having reacted with the (Figure 3). It doesn’t matter which type of protein is used to create the standard curve because the BCA reagent reacts the same with all protein, so we will use a protein called bovine serum albumin (BSA). The slope (m) of the line (y = mx + b) estimates the absorbance coefficient for the BCA reagent after reacting with the protein. The slope can then be used to calculate the concentration of an unknown from it’s absorbance as: c = Abs ) m. Figure 3. A BCA reagent standard curve is prepared by measuring the absorbance of samples with known amounts of BSA.
俄罗斯分光光度计PHOTON RT说明书
PHOTON RTUniversal Scanning SpectrophotometerOperation ManualPKTH.033.000.0001IMPORTANT NOTICECopyright InformationThis document contains proprietary information that is protected by copyright. All rights are reserved. Neither the whole document nor any part of this document may be reproduced in any form or by any means or translated into any language without the prior and written permission of EssentOptics Ltd. Copyright © 2012-2016 EssentOptics Ltd.TrademarksAll brand names, trademarks, etc. used in this document, even when not specifically marked as such, are protected by law. EssentOptics and PHOTON RT are trademarks of EssentOptics Ltd.Contents1 SAFETY MEASURES (4)2 DESCRIPTION AND OPERATION OF PHOTON RT SPECTROPHOTOMETER (5)2.1 P URPOSE (5)2.2 P RODUCT S PECIFICATIONS (6)2.3 C OMPLETE SET OF SPECTROPHOTOMETER (6)2.4 C ONFIGURATION OF SPECTROPHOTOMETER (7)2.5 M ARKING AND SEALING (9)2.6 P ACKING (9)3 INSTALLATION (10)3.1 P REPARING FOR OPERATION (10)3.2 P HOTON S OFT I NSTALLATION (11)3.2.1 PC REQUIREMENTS (11)3.2.2 S OFTWARE INSTALLATION (11)3.3 O PERATION OF THE SPECTROPHOTOMETER (12)3.3.1 M EASUREMENT OF TRANSMITTANCE. (13)3.3.2 M EASUREMENT OF ABSOLUTE SPECULAR REFLECTANCE. (14)3.3.3 M EASUREMENT OF ABSORPTANCE SPECTRA (15)3.3.4 P OLARIZATION-DEPENDENT MEASUREMENTS AT VARIABLE ANGLES IN PS MODE. (16)3.3.5 T RANSMITTANCE MEASUREMENT OF THICK SAMPLES AT HIGH ANGLES OF INCIDENCE. (18)3.3.6 M EASUREMENT OF COMPLEX REFRACTIVE INDEX AND LAYER THICKNESS (19)3.3.7 B ATCH MEASUREMENTS (21)3.3.8 V ERIFICATION OF WAVELENGTH CALIBRATION OF THE SPECTROPHOTOMETER (22)3.4 P HOTON S OFT S OFTWARE (24)3.4.1 C ONTROL COMMANDS (24)3.4.2 S ETTINGS OF MEASURING PARAMETERS (26)3.4.3 S PECTRAL GRAPHS (27)3.4.4 S AVING OF SPECTRA (28)3.4.5 O PTICAL DENSITY (29)3.4.6 K INETIC MEASUREMENT (29)3.4.7 P RINTING OF REPORT (32)3.4.8 B EAM DISPLACEMENT CALCULATOR (36)3.4.9 M EASUREMENT OF COMPLEX REFRACTIVE INDEX AND LAYER THICKNESS. (37)3.4.10 M ETHODS (39)3.4.11 B ATCH MEASUREMENTS (40)3.4.12 I NTERFACE SETTINGS (44)4 MAINTENANCE AND REPAIRS (46)4.1 R EPLACEMENT OF LIGHT SOURCES (46)4.1.1 R EPLACEMENT OF HALOGEN LAMP (46)4.1.2 R EPLACEMENT OF DEUTERIUM LAMP (48)5 STORAGE (50)6 TRANSPORTATION (50)7 UTILIZATION (50)8 ACCEPTANCE CERTIFICATE (51)9 PACKING CERTIFICATE (52)10 WARRANTY (53)11 APPENDIX 1. PRODUCT SPECIFICATION (54)12 APPENDIX 2. COMPLETE SET OF SPECTROPHOTOMETER (56)13 APPENDIX 3. WAVELENGTH CALIBRATION TABLE (57)1Safety measuresThe spectrophotometer complies with the safety standard IР STB 14254-96 standard.Prior to the operation, read the safety rules and regulations for electric equipment carefully and follow the necessary instructions for operation of the spectrophotometer. One should be clearly aware of danger of hazardous internal and external voltages.CAUTION!It is PROHIBITED to operate the spectrophotometer with the removed housing.It is PROHIBITED to operate the spectrophotometer after ingress of water.It is PROHIBITED to operate the spectrophotometer without proper grounding.2Description and operation of Photon RT spectrophotometer2.1PurposeThe Photon RT universal scanning spectrophotometer (further referred to as spectrophotometer) is designed to measure optical characteristics of absolute specular reflectance, transmittance, absorptance, and optical density of planar optical samples with thin film coatings at variable angles and in the polarized light.The spectrophotometer is developed on the basis of the Czerny-Turner monochromator. The spectrophotometer is intended for operation in laboratory conditions in accordance with the following requirements:∙ambient temperature – from +10 0С to +28 0С;∙relative humidity – below 80 % at the temperature + 25 оС;∙atmospheric pressure – from 84 kPa to 106.7 kPa∙proper grounding at the connection point of spectrophotometer and computer2.2Product SpecificationsSee Appendix 1for specifications of your individual spectrophotometer.2.3Complete set of spectrophotometerSee Appendix 2 for complete set of your individual spectrophotometer.2.4Configuration of spectrophotometerFigure 2.4.1 shows the spectrophotometer (front view) with an open lid.The lid of the measuring compartment 1 ensures protection against ambient illumination. In the process of measurement a sample is placed on the sample holder 3 of the sample stage 2 and is fastened with clamps 4. When the spectrophotometer is switched on, red-color flickering of indicator light-emitting diode (LED) 7 continues until the end of the spectrophotometer initialization and self-check procedure. When the spectrophotometer is in the off-measuring mode (“ready” mode), flickering of the indicator LED is green. The indicator LED 7 flickers red during the measurement process, and red-yellow during the change of diffraction gratings. The spectrophotometer is switched on and off by switch 6.Photodetector unit 5 is positioned at a supporting holder of the photodetector drive. The drive provides for positioning of photodetectors along the optical axis for the angles from 16˚ to 183˚: in the case of reflectance measuring -synchronously or non-synchronously with the stage rotation, in the case of transmittance measuring - photodetectors are positioned at normal angle of incidence or any angle within 0˚ - 75˚ range selected by the user.Rotation of stage 2 in the horizontal plane around the optical axis is realized from 0˚ to 75˚. Both rotations are executed with 0,10 step.The value of the incidence angle on the measured surface is set in the field «Measuring parameters» (see Subsection 2.4.2). The position of photodetector is adjusted automatically with the measuring mode («TRANSMITTANCE» or «REFLECTANCE») and depending on the rotation angle of sample stage 2. Rotation angles of sample stage 2 and photodetector unit 5 can be also realized independently in the manual mode for measurement of complex prismatic units. The motorized displacement of photodetector unit perpendicular to the axis of the light beam allows measuring the transmittance of thick samples at high angles up to 75˚. Operation of the beam displacement calculator is described in section 2.4.For the adequate measurement of reflectance, the sample’s surface must be pressed to the stage surface. The measuring area is at the opening center of stage 2.1. Lid of measuring compartment;2. Sample stage;3. Holder;4. Clamps;5. Photodetector unit;6. Power switch;7. Indicator LED;Figure 2.4.1 Photon RT spectrophotometer, front view, with open lid.Figure 2 shows a rear view of the spectrophotometer. The rear panel has USB connector 1 for connection of PC, power supply connector 2, and fuse 3 (including the spare fuse inside). Cover 4 of the compartment with light sources is fastened to the side surface of the instrument by screws 5. The procedure for replacement of the light sources is described in Subsection 4.11. USB connector;2. Power supply connector;3. Fuse (including spare fuse inside);4. Cover;5. Screws.Figure 2.4.2 Photon RT spectrophotometer, rear view.2.5Marking and sealingMarking of the spectrophotometer includes:∙brand name (description) of the device;∙manufacturer’s trademark;∙legends for the elements of connection to external devices are given on the rear panel of the spectrophotometer;∙serial number of the tool is provided on the rear panel;∙marking on the shipment package2.6PackingThe spectrophotometer is packed in accordance with the manufacturer’s requirements and specifications.3Installation3.1Preparing for operation1)Open the packing container, take out the operation manual (OM), take out the spectrophotometer. Whentransportation of the spectrophotometer is handled at temperature below 5оС, leave it unpacked for no less than 24 hours.2)Install the spectrophotometer on the solid horizontal surface.3)Check for the set completeness (see Subsection 2.3).4)Inspect the instrument to make sure that there is no mechanical damage.5)Install «PhotonSoft» Software using PC in accordance with Subsection 3.2.6)Provide effective grounding of the spectrophotometer and PC.7)Connect the spectrophotometer to power mains using the power cable.8)Connect the spectrophotometer to PC with the help of USB cable (see Figure 2.4.2).9)Switch-on the spectrophotometer.10)Start «PhotonSoft» Software.IMPORTANT NOTE:After “PhotonSoft” software starts, the instrument runs self-check and initialization procedure of all controlled elements. This may take about 1 minute.It is PROHIBITED to switch off / switch on the instrument, close / open the software or activate any elements of the software during the self-check and initialization procedure. Otherwise this may damage the instrument.3.2PhotonSoft Installation3.2.1PC requirements∙Microsoft Windows XP/ Windows7/Windows 8operating system;∙SVGA monitor with the resolution no less than 1024x768 (optimum 1280х1024) dots;∙Video adapter memory capacity no less than 32Мb (optimum 64Мb) and color depth no less than 16 bit; ∙No less than 128 Мb of memory (RAM);∙Keyboard, mouse-type manipulator;∙Hard disk (HDD) with free capacity no less than 10 Gb.3.2.2 Software installationTo install the Software, perform the following operations:∙make sure the spectrophotometer is NOT connected to PC∙insert the Software CD-disk (or USB-flash) into CD-ROM (or USB port) of PC ;∙start the installation software setup.exe and follow its instructions;∙press the «Next» button in the installation window ;∙press «Next» in the installation window;∙after successful installation of the software, press the «Finish» button in the window;∙find quick start tag of «PhotonSoft» on the desktop of your PC;∙connect the spectrophotometer to PC;The spectrophotometer is ready for operation.3.3Operation of the spectrophotometer3.3.1Measurement of transmittance.∙Switch on the spectrophotometer.∙Start «PhotonSoft» from your desktop.IMPORTANT NOTE:After “PhotonSoft” software starts, the instrument runs self-check and initialization procedure of all controlled elements. This may take about 1 minute.It is PROHIBITED to switch off / switch on the instrument, close / open the software or activate any elements of the software during the self-check and initialization procedure. Otherwise this may damage the instrument.∙The warm-up time of the spectrophotometer shall be no less than 30 minutes.∙Select «TRANSMITTANCE» from the drop-down menu.∙Set the necessary measuring parameters (see Subsection 3.4.1).1) scanning range;2) sampling pitch;3) averaging count;4) smoothing mode;5) sample stage angle setting;6) polarization.∙Open the lid and make sure that the optical channel in the measuring compartment has no objects.∙Close the lid.∙Press the button «Apply».∙Press the button «Calibration» for baseline calibration. When scanning of a spectrum is finished, the screen displays the horizontal spectral graph serving as 100% transmittance level.∙Open the lid. Place a sample to be measured on the sample stage. Close the lid.∙Start the measuring process by pressing the button «MEASUREMENT». When the process is finished, the screen displays the graph for a transmittance spectrum of the sample.IMPORTANT NOTES:1)When starting the instrument on the new day, it is recommended to repeat baseline calibration beforeactual measurements for breaking-in after 30 minutes warm-up time. This ensures small bit adjustments of the moving parts after idle time.Recommended parameters:∙Staring wavelength: 400 nm;∙Ending wavelength: 1600 nm;∙Sampling pitch: 10 nm;∙Averaging count: 10;∙Smoothing mode: 0;2)Select the wavelength scanning range applicable to the measured sample.3)For more precise measurements in UV-VIS or VIS range, set scanning range and run baselinecalibration up to 990 nm. (For example, 180-990 nm or 380-990 nm respectfully.)4)For more precise measurements in IR range, set scanning range and run baseline calibration starting1000 nm. (For example, or 1000-1600 nm or 1000-3000 nm etc).5)When measurements are conducted over the complete effective wavelength range of thespectrophotometer, it is recommended to perform the baseline calibration directly before measuring the spectrum of the sample.Figure 3.3.1 Transmittance and optical density spectrum.Figure 3 illustrates transmittance and optical density spectrum. The transmittance scale is on the left and the optical density scale is on the right. The optical density graph may be displayed or hidden (see Subsection2.4.4).3.3.2Measurement of absolute specular reflectance.∙Switch on the spectrophotometer.∙Start «PhotonSoft».IMPORTANT NOTE:After “PhotonSoft” software starts, the instrument runs self-check and initialization procedure of all controlled elements. This may take about 1 minute.It is PROHIBITED to switch off / switch on the instrument, close / open the software or activate any elements of the software during the self-check and initialization procedure. Otherwise this may damage the instrument.∙The warm-up time of the spectrophotometer shall be no less than 30 minutes.∙Select «TRANSMITTANCE» from the drop-down menu.∙Set the required measuring parameters (see Subsection 3.4.1)1) scanning range;2) sampling pitch;3) averaging count;4) smoothing mode;5) polarization.∙Make sure that the optical channel in the measuring compartment has no objects.∙Press the button«Apply».∙Press the button «Calibration» for baseline calibration. When scanning of a spectrum is finished, the screen displays the horizontal spectral graph serving as 100% transmittance level.∙Select «REFLECTANCE» from the drop-down menu to change for measurement of absolute specular reflectance.∙Open the lid. Place a sample to be measured on the sample stage. The coated surface shall be facing the sample stage for measurement of absolute specular reflectance. Close the lid.∙Set the angle for sample stage.∙Press the button«Apply».∙Start the measuring process by pressing the button «MEASUREMENT». When the process is finished, the screen displays the graph for the absolute reflectance spectrum of the sample.IMPORTANT NOTES:1)When starting the instrument on the new day, it is recommended to repeat baseline calibration beforeactual measurements for breaking-in after 30 minutes warm-up time. This ensures small bit adjustments of the moving parts after idle time.Recommended parameters:∙Staring wavelength: 400 nm;∙Ending wavelength: 1600 nm;∙Sampling pitch: 10 nm;∙Averaging count: 10;∙Smoothing mode: 0;2)Select the wavelength scanning range applicable to the measured sample.3)For more precise measurements in UV-VIS or VIS range, set scanning range and run baselinecalibration up to 990 nm. (For example, 180-990 nm or 380-990 nm respectfully.4)For more precise measurements in IR range, set scanning range and run baseline calibration starting1000 nm. (For example, 1000-3000 nm, 1000-1650 nm etc.)5)When measurements are conducted over the complete effective wavelength range of thespectrophotometer, perform baseline calibration directly before measuring the spectrum of the sample.3.3.3Measurement of absorptance spectraThe Photon RT spectrophotometer provides the possibility to measure absorptance spectra of the unknown transparent substrate. The measurement of the absorptance spectra is realized by sequential measurements of transmittance and reflectance, and subsequent processing of the measurement results.∙Switch on the spectrophotometer.∙Start «PhotonSoft».IMPORTANT NOTE:After “PhotonSoft” software starts, the instrument runs self-check and initialization procedure of all controlled elements. This may take about 1 minute.It is PROHIBITED to switch off / switch on the instrument, close / open the software or activate any elements of the software during the self-check and initialization procedure. Otherwise this may damage the instrument.∙The warm-up time of the spectrophotometer shall be no less than 30 minutes.∙Select «TRANSMITTANCE» from the drop-down menu.∙Set the required measuring parameters (see Subsection 3.4.1)1) scanning range;2) sampling pitch;3) averaging count;4) smoothing mode;5) polarization.∙Make sure that the optical channel in the measuring compartment has no objects.∙Press the button«Apply».∙Press the button «Calibration» for baseline calibration. When scanning of a spectrum is finished, the screen displays the horizontal spectral graph serving as 100% transmittance level.∙Select «ABSORPTANCE» from the drop-down menu to change for measurement of absorptance.∙Place a sample to be measured on the sample stage.NOTE: One can use the same sample to measure both transmittance and reflectance, if the sample has thickness of 40 mm and above. Otherwise, for reflectance measurementone should prepare and use a 50 wedge sample made of the same material.∙Input the value of the sample thickness in the line “Sample thickness, mm” (See Fig. 3.4.4).∙Press the button«Apply».∙Start the measuring process by pressing the button «MEASUREMENT». When the process is finished, the screen displays the graph for the transmittance spectrum of the sample. After that, thedetectors unit and sample stage will synchronously rotate for reflectance measurement at 80.∙Start the measuring process by pressing the button «MEASUREMENT». When the process is finished, the screen displays the graph for the internal attenuation DA which represents the value of total internal losses for absorptance and scattering of the signal in the measured sample.3.3.4Polarization-dependent measurements at variable angles in PS mode.The spectrophotometer has built-in high-contrast polarizers that operate unattended. This configuration provides for polarization-dependent measurement of transmittance, absolute specular reflectance at variable angles of incidence, and measurement/calculation of optical constants (refractive index, layer thickness and extinction coefficient).During the PS mode of measurement, the spectrum is measured subsequently for S polarization and for P polarization without any involvement of operator. Next, the (S+P)/2 value of random polarization is calculated and displayed instantly for transmittance or absolute specular reflectance.∙Switch on the spectrophotometer.∙Start the «PhotonSoft».IMPORTANT NOTE:After “PhotonSoft” software starts, the instrument runs self-check and initialization procedure of all controlled elements. This may take about 1 minute.It is PROHIBITED to switch off / switch on the instrument, close / open the software or activate any elements of the software during the self-check and initialization procedure. Otherwise this may damage the instrument.∙The warm-up time of the spectrophotometer shall be no less than 30 minutes.∙Set the required measuring parameters (see Subsection 3.4.1)(1) scanning range;(2) sampling pitch;(3) averaging count;(4) smoothing mode;(5) polarization - PS.(6) slid width∙Open the lid, make sure that the optical channel in the measuring compartment has no objects. Close the lid.∙Press the button«Apply».∙Press the button «CALIBRATION».∙Open the lid. Place a sample to be measured on the sample stage. The coated surface shall be facing the sample table for measurement of reflectance. Close the lid.∙Select «TRANSMITTANCE» or “REFLECTANCE” measurement mode from the drop-down menu. ∙Set the angel for the sample stage.∙Press the button«Apply».∙To start the measuring process, press the button «MEASUREMENT».In the «PS» polarization mode the baseline calibration is performed twice: in «S» position of the polarizer and in «P» position of the polarizer. The displayed spectrum for «S» polarization is dark blue and that for «P» polarization is dark green in color. The calculation of average polarization «(S+P)/2» is performed after completion of subsequent measurements for the spectra associated with «S» and «P» polarizations. The resultant spectrum is displayed in white color on the screen immediately.∙When the process is finished, the screen displays a transmittance or reflectance spectrum at the specified angle of incidence at (S+P)/2 average polarization.IMPORTANT NOTES:1)When starting the instrument on the new day, it is recommended to repeat baseline calibration beforeactual measurements for breaking-in after 30 minutes warm-up time. This ensures small bit adjustments of the moving parts after idle time.Recommended parameters:2)Staring wavelength: 400 nm;3)Ending wavelength: 1600 nm;4)Sampling pitch: 10 nm;5)Averaging count: 10;6)Smoothing mode: 0;7)Select the wavelength scanning range applicable to the measured sample.8)For more precise measurements in UV-VIS or VIS range, set scanning range and run baselinecalibration up to 990 nm. (For example, 180-990 nm or 380-990 nm respectfully.)9)For more precise measurements in IR range, set scanning range and run baseline calibration starting1000 nm. (For example, 1000-3000 nm, 1000-1650 nm etc.)10)W hen measurements are conducted over the complete effective wavelength range of thespectrophotometer, perform calibration directly before measuring the spectrum for a sample.11)S elect the slit width approximately 1.5 times bigger compared to regular (not PS) measurementprocedure. Make sure the maximum signal value does not exceed 65 000 units after baseline calibration (refer to the Signal window of the main interface).Figure 3.3.2 Transmittance spectrum in «PS» mode.Figure 3.3.2 shows a transmittance spectrum of the optical coating at the 45˚ angle of incidence in «PS» mode. The spectra for «S» and «P» polarizations are displayed in darkened colors.3.3.5Transmittance measurement of thick samples at high angles of incidence.When measuring transmittance of thick optical samples at high angles of incidence, the parallel displacement of transmitted beam occurs. The value of beam displacement depends on three factors: angle of incidence, physical thickness of the sample and refractive index of the sample material.The PHOTON RT spectrophotometer offers a possibility for correct transmittance measurement of thick sample at high angles using the built-in “Beam displacement calculator” option (See also Section 3.4.8). The actual value of beam displacement can be within 0-10 mm range.∙Switch on the spectrophotometer.∙Start the «PhotonSoft».IMPORTANT NOTE:After “PhotonSoft” software starts, the instrument runs self-check and initialization procedure of all controlled elements. This may take about 1 minute.It is PROHIBITED to switch off / switch on the instrument, close / open the software or activate any elements of the software during the self-check and initialization procedure. Otherwise this may damage the instrument.∙The warm-up time of the spectrophotometer shall be no less than 30 minutes.∙Set the required measuring parameters (see Subsection 3.4.1)(1) scanning range;(2) sampling pitch;(3) averaging count;(4) smoothing mode;(5) sample table angle setting(6) polarization - PS.∙Open the lid. Make sure that the optical channel in the measuring compartment has no objects. Close the lid.∙Press the button«Apply».∙Press the button «Calibration» for baseline calibration. When scanning of a spectrum is finished, the screen displays the horizontal spectral graph serving as 100% transmittance level.∙Open the lid. Place a sample to be measured on the sample stage.∙Select desired angle of incidence. Close the lid.∙Select «BEAM DISPLACEMENT CALCULATOR» from the drop-down menu «Tools”. The “Beam displacement calculator” window opens (see Figure 3.4.21).∙In the “Beam displacement calculator” window, fill in appropriate fields “Thickness” (in mm) and “Refractive index” for the measured sample. The angle of sample table is uploaded from the “Sample table angle setting” field (see Figure 3.4.4).∙The value of beam displacement (in mm) is calculated immediately.∙Press the button«APPLY» in the “Beam displacement calculator” window. The value of beam displacement (in mm) will be immediately displayed in the “Detector displacement, mm” field of the “Display options” menu (see Figure 3.4.4).∙Press the button«APPLY» in the “Display options” menu (see Figure 3.4.4).∙Start the measuring process by pressing the button «MEASUREMENT». When the process is finished, the screen displays the graph for transmittance spectra of the sample at a user-selected angle of incidence.∙Change the beam displacement value for “0” in the “Detector displacement, mm” field of the “Display options” menu (see Figure 3.4.4) if no further transmittance measurements of thick samples at highangles of incidence are required.∙Press the button«APPLY» in the “Display options” menu (see Figure 3.4.4).The beam displacement of the detectors can also be realized by setting the check box in the field "Detector autodisplacement" (see. Figure 3.4.3). The following lines become active: "Sample thickness, mm" and "Refractive index n”. The value of the sample thickness and the average refractive index of the sample material for the selected wavelength range shall be indicated in the appropriate fields. After clicking the "Apply" button, the value of detector displacement will be displayed in the "Detector displacement, mm" field. The detectors will immediately move at the calculated value of displacement.3.3.6Measurement of complex refractive index and layer thicknessOptical constants of the material layers are characterized with a number of parameters. Several of them are important for experts involved in optical design of the multilayer coatings – refractive index (n), layer thickness (d) and extinction coefficient (k).The NKD calculation software is designed to calculate the refractive index (n), extinction coefficient (k) and layer thickness (d) of the single homogenous layers on the known substrate using the photometric reverse engineering method. Calculations of optical parameters of the thin film layers is based on measuring the reflectance of the coated substrate in a polarized light (Rp and Rs values) for several angles of incidence. Typically, 3 to 8 angles of incidence are sufficient for correct measurement and calculation. Recommended sampling pitch is from 5 to 20 nm. The wedged substrate (with not less than 5 deg wedge angle) must be used for correct measurement to exclude reflectance from the back surface. BK7 and SiO2 substrates are recommended as test substrates for described measurement. It is necessary to increase the slid with 1,5 times to compensate for decreased signal in the PS measurement mode.∙Switch on the spectrophotometer.∙Start the «PhotonSoft».。
吸收光谱简介 Absorption Spectrum An Introduction 英语作文论文
吸收光谱简介Absorption Spectrum AnIntroductionWhat is Absorption Spectrum?The absorption spectrum of a material can be defined as the fraction of the incident radiation which is absorbed by that material over a wide range of frequencies. The molecular and atomic composition of a material is used to determine the absorption spectrum. Fundamental radiation is generally observed at those frequencies which get mixed with the energy difference that takes place between two mechanical states of the molecules.The absorption takes place because of the transition between these two states and it is known as the absorption line. The spectrum is composed of several absorption lines. The frequencies where such absorption lines develop along with their relative intensities generally depend on the molecular structure and electronic structure of the sample. The frequencies also depend on molecular interactions. In the sample, the crystal structure is found in solids and on different environmental factors like pressure, temperature, electromagnetic fields, etc.What is Absorption Spectrum?Assingment Experts will explain Absorption Sepctrum in deatils. The absorption spectrum of a material can be defined as the fraction of theincident radiation which is absorbed by that material over a wide range of frequencies. The molecular and atomic composition of a material is used to determine the absorption spectrum. Fundamental radiation is generally observed at those frequencies which get mixed with the energy difference that takes place between two mechanical states of the molecules.The absorption takes place because of the transition between these two states and it is known as the absorption line. The spectrum is composed of several absorption lines. The frequencies where such absorption lines develop along with their relative intensities generally depend on the molecular structure and electronic structure of the sample. The frequencies also depend on molecular interactions. In the sample, the crystal structureis found in solids and on different environmental factors like pressure, temperature, electromagnetic fields, etc.The absorption lines also possess a definite shape and width which are fundamentally determined by the density of states for spectral density of the system. Absorption lines are generally classified by the feature of quantum mechanical change taking place in an atom or molecule. Rotational states sometimes get changed and give result in the development of rotational lines which are found in the region of the microwave spectrum. On the other hand, vibrational lines in correspondence to vibrational state changes in the molecule are found in the area of infrared region. The electronic lines are composed of several changes taking place in the electronic state of a molecule or atom whichare found in the ultraviolet and visible region.It can be noticed that there are various dark lines in the sun’s spectrum. These lines are developed by the atmosphere of the Sun which absorbs light at different wavelengths resulting in different light intensity at the wavelength to appear dark. The molecules and atoms present in a gas absorb certain light wavelengths. The pattern of the lines is very unique with respect to each element which provides us information about the elements which help in making the sun’s atmosphere. The absorption spectra can be observed from spatial regions in the presence of a cooler gas line between in a hotter source and the earth.The absorption spectra can also be observed from the planets with atmospheres, stars, and galaxies. In analyzing the light of the Sun, aspectrometer is used. The spectrometer is a device which separates light by colour and energy. In separating light by colour and energy, the image of the spectrum of the sun gets created. This is quite similar to the absorption spectrum. The dark lines are the areas where the light gets absorbed by different elements present in the Sun’s outer layers. The lowest energy is represented by red light and the highest energy is represented by blue light.The black gaps or lines in the spectrum of the sun are termed as absorption lines. The gas present in the sun’s outer layers develops the absorption lines by absorbing the light. There are different elements such as Helium, hydrogen, carbon, and other smaller quantities of heavy elements in the sun. When the sunlight shines, the elements the energy gets absorbed by the atoms. The atoms can only absorb the lightrelevant to the energy the atoms need. The gaps in the spectrum of the Sun get developed and help in informing the formation of the sun. The emission spectrum is quite different from the absorption spectrum.In developing an absorption spectrum, the light needs to shine through a gas but in creating and emission spectrum a gas needs to be heated up. The atoms present in the gas get absorbed the energy only for a short tenure. The atoms get energetic and jiggled up by heating the gas because of the concentration of a high level of energy. The energy is emitted or re-released as light eventually. Absorption spectrum takes place when the light passes through a dilute and cold gas and characteristic frequencies get absorbed by the atoms present in the gas. The re-emitted light cannot be emitted in a similardirection which is followed by absorbed Photon because of which dark lines in the spectrum are created in the absence of light. The absorption spectrum is the dark lines. The absorption spectrum is defined as an Electromagnetic Spectrum in which the radiation intensity at some specific wavelengths gets decreased. An absorbing substance gets manifested as bands or dark lines. Medically, the absorption spectrum is also defined as an Electromagnetic Spectrum in which radiation intensity at specific ranges of wavelength is manifested as dark lines.X-ray absorptions are highly associated with the excitation taking place in the inner shell electrons in an atom. These changes generally get combined to develop a new absorption line which is typically found in the combined energy develop mainly during the changes. The changes are mainly radiation-vibrationstransitions. The energy which is typically found in the quantum mechanical change fundamentally determines the absorption line frequency. The frequency can get shifted because of several interactions. The magnetic and electric fields can give result in a shift.The interactions with some of the neighbouring molecules can also cause shifts. Absorption lines of any gas-phase molecule can get shifted typically when the molecule is present in either solid or liquid phase and involves in interacting with neighbouring molecules strongly. The shape and width of the absorption lines are generally determined by the observation instrument. The physical environment radiation and material absorbing of that material also determine the shape and width of absorption lines. Now our experts from Instant AssingnmentHelp will tell you about the relationship between Absorption Spectrum andThe relation between Transmission and absorption spectraTransmission and absorption spectra are interconnected. Transmission and absorption spectra are found to represent similar information. Transmission spectrum can be calculated from the absorption spectrum only. Absorption spectrum can also be calculated from transmission spectra. Mathematical transformation is used in calculating either the absorption spectrum or transmission spectrum. It has been observed that a transmission spectrum has maximum intensities where thewavelengths of the absorption spectrum are quite weak because of the transmission of more light through the sample takes place. Similarly, an absorption spectrum is found to have maximum intensities at its wavelengths where the absorption rate is quite stronger.The absorption spectrum is also related to any emission spectrum. Now, it is important to understand the concept of the emission spectrum. The process by which a substance can release energy is known as emission process. The energy which is released from a substance through any emission process can be found in electromagnetic radiation from. Emission can take at any frequency of absorption which makes the absorption lines to gets determined from the emission spectrum. But it is to be remembered that the emissionspectrum will always have different intensity pattern where it becomes distinguished from that of the absorption spectrum. Hence, it can be said that the absorption spectrum and emission spectrum can never be equivalent. The emission spectrum can be used to calculate the absorption spectrum with the application of effective theoretical models and other relevant information from where quantum mechanical states of a substance can be understood.Relationship between Absorption spectrum and reflection and scattering spectraThe absorption spectrum is also related to reflection and scattering spectra. The scattering and reflection spectra of any material getinfluenced by the absorption spectrum and index of refraction of that material. Extinction coefficient quantifies the absorption spectrum and index coefficients along with extinction coefficients which are related through Kramers-Kroening relation quantitatively. Therefore, it can be said that reflection or scattering spectrum standardize absorption spectrum can give rise to absorption spectrum.Reflection or scattering spectrum assumptions or models need to be simplified so that it can lead to effect an approximation of the derivation of absorption spectra. In the domain of chemical analysis, we can find the use of absorption spectroscopy because of the quantitative nature and specificity of the absorption spectrum. The specificity enables the compounds to get distinguished from each other in a mixture whichmakes absorption spectroscopy to be highly useful in different applications. For example, the presence of any pollutant in the air can be identified by the use of infrared gas analyzers.These analyses are also used to distinguish the air pollutant from oxygen, water, nitrogen, and other constituents. The specificity is also helpful in allowing several unknown samples to get rightly identified. It can be done by comparing the measured spectrum with the findings of reference spectra. It has been found that qualitative information of any sample can also be determined even if the information is not present in a library. For example, infrared spectra have several characteristics absorption bands which help in indicating the presence of carbon-oxygen bond or Carbon hydrogen bonds.Absorption spectrum can also be related to the quantity of material present with the use of Beer-Lambert law. This relationship is established quantitatively. In determining the typical compound concentration, it needs knowledge of the absorption coefficient of the compound. The absorption coefficient can be known from several reference sources and can be measured by accessing calibre standard spectrum with an available target concentrationabsorption spectrumAbsorption spectroscopy and its applicationAbsorption spectroscopy is one of the methods with the help of which a substance can get characterized by the support of wavelengths at which the spectrum of colour gets absorbedduring the passage of light through a substance solution. It is one of the fundamentally used methods used in assessing the chromospheres concentrations in the solutions. Absorption spectroscopy can also be explained as a non-destructive technique which is widely used by biochemists and biologists to assess the characteristic parameters and cellular components of functional molecules.This quantification is highly important in the domain of systems biology. In developing metabolic pathway quantitative depiction, various variables and parameters are needed which are to be assessed experimentally. Ultraviolet-visible absorption spectroscopy is used in producing experimental data which help in modelling techniques of system biology. These techniques use kinetic parameters andconcentrations of enzymes of signalling on metabolic pathways, fluxes, and intercellular metabolic concentrations. Absorption spectroscopy also describes the usage of the technique in quantifying bio-molecules and investigating bio-molecular interactions.Absorption spectroscopy is a significant technique which is used in chemistry to study simple inorganic species. It refers to spectroscopic techniques which are used in measuring radiation absorption as a function of wavelength or frequency when the interaction between absorption radiation and sample takes place. Photons are absorbed by the samples from the field of radiation. The absorption intensity varies as a frequency function and this absorption intensity is the absorption spectrum. Absorption spectroscopy is fundamentallyperformed across an absorption spectrum or electromagnetic spectrum.In the domain of analytical chemistry, absorption spectroscopy is used to assess the presence of any specific substance in a sample. In several cases, absorption spectroscopy is also used to quantify the quantity of a substance. In the domain of analytical applications, ultraviolet-visible and infrared spectroscopy is commonly observed. In the study of atomic physics, remote sensing, molecular physics, and astronomical spectroscopy, the use of absorption spectroscopy are widely observed.There are various experimental approaches which are used to measure the absorption spectrum. The most commonly used arrangement is to guide the regeneratedradiation beam at the sample in detecting the radiation intensity passing through it. The transmitted energy can be applied in calculating the absorption. The sample arrangement source and detection technique are also very used quite significantly depending on the objective of the experiment and that of the frequency range.Advantages of absorption spectroscopyThere can be several advantages of absorption spectroscopy because it can be used as an analytical method where measurements can be accomplished without any contact between the sample and the instrument. Radiation which travels between an instrument and a sample contains some important spectral information and measurement which is done remotely. Remote spectral sensing is quite significant indifferent situations. For example, hazardous and toxic environments can be measured without risking any instrument or operator.The material of the sample needs not to be brought into direct contact with any instrument which can prevent cross-contamination at a possible rate. Remote spectral measurements have certain challenges as compared to that of the laboratory measurements. To reduce such challenges, differential optical absorption spectroscopy has become quite popular because it mainly emphasizes on the features of differential absorption and erasers broadband absorption like the extinction of aerosol extinction because of Rayleigh scattering. This technique is used in airborne, ground-based, and satellite-based measuring actions. There are certain ground-based techniques whichprofile the possibilities of retrieving stratospheric and tropospheric trace gas profiles.The absorption lines also possess a definite shape and width which are fundamentally determined by the density of states for spectral density of the system. Absorption lines are generally classified by the feature of quantum mechanical change taking place in an atom or molecule. Rotational states sometimes get changed and give result in the development of rotational lines which are found in the region of the microwave spectrum. On the other hand, vibrational lines in correspondence to vibrational state changes in the molecule are found in the area of infrared region. The electronic lines are composed of several changes taking place in the electronic state of a molecule or atom which are found in the ultraviolet and visible region.Itcan be noticed that there are various dark lines in the sun’s spectrum. These lines are developed by the atmosphere of the Sun which absorbs light at different wavelengths resulting in different light intensity at the wavelength to appear dark. The molecules and atoms present in a gas absorb certain light wavelengths. The pattern of the lines is very unique with respect to each element which provides us information about the elements which help in making the sun’s atmosphere. The absorption spectra can be observed from spatial regions in the presence of a cooler gas line between in a hotter source and the earth.The absorption spectra can also be observed from the planets with atmospheres, stars, and galaxies. In analyzing the light of the Sun, a spectrometer is used. The spectrometer is adevice which separates light by colour and energy. In separating light by colour and energy, the image of the spectrum of the sun gets created. This is quite similar to the absorption spectrum. The dark lines are the areas where the light gets absorbed by different elements present in the Sun’s outer layers. The lowest energy is represented by red light and the highest energy is represented by blue light.The black gaps or lines in the spectrum of the sun are termed as absorption lines. The gas present in the sun’s outer layers develops the absorption lines by absorbing the light. There are different elements such as Helium, hydrogen, carbon, and other smaller quantities of heavy elements in the sun. When the sunlight shines, the elements the energy gets absorbed by the atoms. The atoms can only absorb the light relevant to the energy the atoms need. The gapsin the spectrum of the Sun get developed and help in informing the formation of the sun. The emission spectrum is quite different from the absorption spectrum.In developing an absorption spectrum, the light needs to shine through a gas but in creating and emission spectrum a gas needs to be heated up. The atoms present in the gas get absorbed the energy only for a short tenure. The atoms get energetic and jiggled up by heating the gas because of the concentration of a high level of energy. The energy is emitted or re-released as light eventually. Absorption spectrum takes place when the light passes through a dilute and cold gas and characteristic frequencies get absorbed by the atoms present in the gas. The re-emitted light cannot be emitted in a similar direction which is followed by absorbed Photonbecause of which dark lines in the spectrum are created in the absence of light. The absorption spectrum is the dark lines. The absorption spectrum is defined as an Electromagnetic Spectrum in which the radiation intensity at some specific wavelengths gets decreased. An absorbing substance gets manifested as bands or dark lines. Medically, the absorption spectrum is also defined as an Electromagnetic Spectrum in which radiation intensity at specific ranges of wavelength is manifested as dark lines.X-ray absorptions are highly associated with the excitation taking place in the inner shell electrons in an atom. These changes generally get combined to develop a new absorption line which is typically found in the combined energy develop mainly during the changes. The changes are mainly radiation-vibrations transitions. The energy which is typically foundin the quantum mechanical change fundamentally determines the absorption line frequency. The frequency can get shifted because of several interactions. The magnetic and electric fields can give result in a shift.The interactions with some of the neighbouring molecules can also cause shifts. Absorption lines of any gas-phase molecule can get shifted typically when the molecule is present in either solid or liquid phase and involves in interacting with neighbouring molecules strongly. The shape and width of the absorption lines are generally determined by the observation instrument. The physical environment radiation and material absorbing of that material also determine the shape and width of absorption lines. Now our experts from Instant AssingnmentHelp will tell you about the relationship between Absorption Spectrum andThe relation between Transmission and absorption spectraTransmission and absorption spectra are interconnected. Transmission and absorption spectra are found to represent similar information. Transmission spectrum can be calculated from the absorption spectrum only. Absorption spectrum can also be calculated from transmission spectra. Mathematical transformation is used in calculating either the absorption spectrum or transmission spectrum. It has been observed that a transmission spectrum has maximum intensities where thewavelengths of the absorption spectrum are quite weak because of the transmission of more light through the sample takes place. Similarly, an absorption spectrum is found to have maximum intensities at its wavelengths where the absorption rate is quite stronger.The absorption spectrum is also related to any emission spectrum. Now, it is important to understand the concept of the emission spectrum. The process by which a substance can release energy is known as emission process. The energy which is released from a substance through any emission process can be found in electromagnetic radiation from. Emission can take at any frequency of absorption which makes the absorption lines to gets determined from the emission spectrum. But it is to be remembered that the emissionspectrum will always have different intensity pattern where it becomes distinguished from that of the absorption spectrum. Hence, it can be said that the absorption spectrum and emission spectrum can never be equivalent. The emission spectrum can be used to calculate the absorption spectrum with the application of effective theoretical models and other relevant information from where quantum mechanical states of a substance can be understood.Relationship between Absorption spectrum and reflection and scattering spectraThe absorption spectrum is also related to reflection and scattering spectra. The scattering and reflection spectra of any material getinfluenced by the absorption spectrum and index of refraction of that material. Extinction coefficient quantifies the absorption spectrum and index coefficients along with extinction coefficients which are related through Kramers-Kroening relation quantitatively. Therefore, it can be said that reflection or scattering spectrum standardize absorption spectrum can give rise to absorption spectrum.Reflection or scattering spectrum assumptions or models need to be simplified so that it can lead to effect an approximation of the derivation of absorption spectra. In the domain of chemical analysis, we can find the use of absorption spectroscopy because of the quantitative nature and specificity of the absorption spectrum. The specificity enables the compounds to get distinguished from each other in a mixture whichmakes absorption spectroscopy to be highly useful in different applications. For example, the presence of any pollutant in the air can be identified by the use of infrared gas analyzers.These analyses are also used to distinguish the air pollutant from oxygen, water, nitrogen, and other constituents. The specificity is also helpful in allowing several unknown samples to get rightly identified. It can be done by comparing the measured spectrum with the findings of reference spectra. It has been found that qualitative information of any sample can also be determined even if the information is not present in a library. For example, infrared spectra have several characteristics absorption bands which help in indicating the presence of carbon-oxygen bond or Carbon hydrogen bonds.Absorption spectrum can also be related to the quantity of material present with the use of Beer-Lambert law. This relationship is established quantitatively. In determining the typical compound concentration, it needs knowledge of the absorption coefficient of the compound. The absorption coefficient can be known from several reference sources and can be measured by accessing calibre standard spectrum with an available target concentrationabsorption spectrumAbsorption spectroscopy and its applicationAbsorption spectroscopy is one of the methods with the help of which a substance can get characterized by the support of wavelengths at which the spectrum of colour gets absorbed during the passage of light through a substancesolution. It is one of the fundamentally used methods used in assessing the chromospheres concentrations in the solutions. Absorption spectroscopy can also be explained as a non-destructive technique which is widely used by biochemists and biologists to assess the characteristic parameters and cellular components of functional molecules.This quantification is highly important in the domain of systems biology. In developing metabolic pathway quantitative depiction, various variables and parameters are needed which are to be assessed experimentally. Ultraviolet-visible absorption spectroscopy is used in producing experimental data which help in modelling techniques of system biology. These techniques use kinetic parameters and concentrations of enzymes of signalling onmetabolic pathways, fluxes, and intercellular metabolic concentrations. Absorption spectroscopy also describes the usage of the technique in quantifying bio-molecules and investigating bio-molecular interactions.Absorption spectroscopy is a significant technique which is used in chemistry to study simple inorganic species. It refers to spectroscopic techniques which are used in measuring radiation absorption as a function of wavelength or frequency when the interaction between absorption radiation and sample takes place. Photons are absorbed by the samples from the field of radiation. The absorption intensity varies as a frequency function and this absorption intensity is the absorption spectrum. Absorption spectroscopy is fundamentallyperformed across an absorption spectrum or electromagnetic spectrum.In the domain of analytical chemistry, absorption spectroscopy is used to assess the presence of any specific substance in a sample. In several cases, absorption spectroscopy is also used to quantify the quantity of a substance. In the domain of analytical applications, ultraviolet-visible and infrared spectroscopy is commonly observed. In the study of atomic physics, remote sensing, molecular physics, and astronomical spectroscopy, the use of absorption spectroscopy are widely observed.There are various experimental approaches which are used to measure the absorption spectrum. The most commonly used arrangement is to guide the regeneratedradiation beam at the sample in detecting the radiation intensity passing through it. The transmitted energy can be applied in calculating the absorption. The sample arrangement source and detection technique are also very used quite significantly depending on the objective of the experiment and that of the frequency range.Advantages of absorption spectroscopyThere can be several advantages of absorption spectroscopy because it can be used as an analytical method where measurements can be accomplished without any contact between the sample and the instrument. Radiation which travels between an instrument and a sample contains some important spectral information and measurement which is done remotely. Remote spectral sensing is quite significant indifferent situations. For example, hazardous and toxic environments can be measured without risking any instrument or operator.The material of the sample needs not to be brought into direct contact with any instrument which can prevent cross-contamination at a possible rate. Remote spectral measurements have certain challenges as compared to that of the laboratory measurements. To reduce such challenges, differential optical absorption spectroscopy has become quite popular because it mainly emphasizes on the features of differential absorption and erasers broadband absorption like the extinction of aerosol extinction because of Rayleigh scattering. This technique is used in airborne, ground-based, and satellite-based measuring actions. There are certain ground-based techniques which。
石河子大学药学院——药分药英专业词汇
absolute error 绝对误差absorbance 吸光度absorptivity 吸光系数accidental error 偶然误差accuracy 准确度activity 活度absorption 吸收adsorption 吸附adsorption chromatography 吸附色谱法affinity chromatography 亲和色谱法aliquot (一)份ambient temperature 室温amphoteric solvent 两性溶剂anionic surfactant titration 阴离子表面活性剂滴定法apparatus 仪器ascending development, descending 上行展开(下行)assay 含量测定asymmetry factor 不对称因子atomic absorption spectrophotometry 原子吸收分光光度法attenuation 衰减acid-base titration 酸碱滴定法anode 阳极cathode 阴极aprotic solvent 非质子溶剂atomic emission spectroscopy 原子发射分光光度法average deviation 平均偏差baseline drift 基线漂移batch (lot) number 批号between day (day to day, inter-day) precision 日间精密度within day (intra-day) precision 日内精密度between run (inter-run) precision 批间精密度within run (intra-run) precision批内精密度bromometry 溴量法bonded phase chromatography (BPC) 键合相色谱calibration curve (graph) 校准曲线calorimetry 量热分析colorimetry 比色法capacity factor 容量因子capillary zone electrophoresis (CZE) 毛细管电泳carboxymethyl cellulose (CMC) 羧甲基纤维素carrier gas 载气cation-exchange resin 阳离子交换树脂check valve 单向阀chemical shift 化学位移chiral separation 手性分离chromatogram 色谱clarity 澄清度compleximetry 配位滴定法conductometry 电导法confidence interval 置信区间content uniformity test 含量均匀度检查correlation coefficient 相关系数coulometric titration 库伦滴定法counter ion (反离子)平衡离子cyclodextrin 环糊精characteristic frequency 特征频率charge transfer 手性传荷chelate compound 螯合物chiral ligand-exchange complexes (CLEC) 手性配体复合物conjugate 缀合物critical micellar concentration (CMC) 临界胶束浓度crosslinking column 交联柱daughter ion (质谱)子离子deactivity 脱活性dead-stop titration 永停滴定法deformation vibration (红外)变形振动deliquescence 潮解deproteinization 去蛋白作用derivative 衍生物derivatization 衍生化desiccant 干燥剂desiccator 干燥器dextrose 葡萄糖diastereoisomer 非对映异构体diazotization 重氮化differential scanning calorimetry (DSC) 差示扫描量热法differential thermal analysis DTA 差热分析differentiating effect 区分效应diffusion 扩散digestion 消解disintegration test 崩解试验dispersion 分散度dissolution test 溶出度检查distilling range 馏程distribution coefficient 分配系数drying to constant weight 干燥至恒重dual-wavelength 双波长direct injection 直接进样direct probe inlet (DPI) 直接进样doublet (d) 双峰efflorescence 风化electrochemical analysis 电化学分析electrode potential 电极电位electron capture detector 电子捕获检测器electron impact ionization 电子轰击离子化electron transition 电子跃迁electrospray 电喷雾eluate 洗脱液elution 洗脱emission spectrum 发射光谱enantiomer 对映异构体end absorption 末端吸收end point (滴定)终点endogenous substance (生物样品中)内源性物质equivalent point 等当点(即化学计量点)excitation 激发expiration date 失效日期external standard methond, internal 外(内)标法extraction 提取,萃取electrokinetic injection 电动进样electromagnetic spectrum 电磁波谱electron donating group 供电子取代基electron withdrawing group 吸电子取代基electroosmosis 电渗electroosmotic flow 电渗流evaporative light scatting detector (ELSD) 蒸发光散射检测器fast atomic bombardment (FAB) 快速原子轰击(离子化)field desorption ionization 场解吸离子化filtration 过滤hydrogen flame ionization detector (FID) 氢火焰离子化检测器fluorescence quenching method 荧光熄灭法fluorometry 荧光测定法fluorescence polarization immunoassay 荧光偏振免疫测定法Fourier-transform 傅里叶变换fraction 馏分gel permeation chromatography 凝胶过滤色谱法geminal coupling 偕耦general notices (药典)凡例good clinical practices (GCP) 药品临床试验质量管理规范good laboratory practices (GLP) 药品非临床试验质量管理规范good manufacturing practices (GMP) 药品生产质量管理规范good supply practices (GSP) 药品经营质量管理规范good agricultural practices (GAP) 中药材种植质量管理规范gradient 梯度grating 光栅gravimetric method 重量法height equivalent to a theoretical plate 理论塔板高度heterogeneous membrane electrode, homogeneous 非均相(均相)膜电极high performance liquid chromatography(HPLC)高效液相色谱法holographic grating 全息光栅hydrolysis 水解hydrophilic, hydrophobic 亲水的、疏水的hydroscopic 吸湿的hyperchromic effect, hypochromic 增(减)色效应identification 鉴别ignition 灼烧in vitro 体外in vivo 体内indeterminate error 偶然误差inductive coupling plasma emission 电感耦合等离子体发射infrared spectrophotometry 红外分光光度法inside diameter, internal diameter (i. d. ) 内径integrator 积分仪intercept 截距interface 接口interference filter 干涉滤光片iodimetry 碘量法iodine value 碘值ion suppression 离子抑制ion-paring agent 离子对试剂isoabsorptive point 等吸光点isocratic elution 等度洗脱isotherm 等温线immobilized phase open tubular column 固定化相开管柱instrumental analysis 仪器分析法intersystem crossing 体系间跨越ion abundance 离子丰度isotopic ion 同位素离子labelled amount (quantity) 标示量Latin-square test 拉丁方试验leveling effect 均化效应limit of detection 检测限limit of quantitation 定量限linearity 线性lipophilic 亲脂的local diamagnetic shielding 局部抗磁屏障loss on drying 干燥失重luminescence 发光lyophilization 冷冻干燥level of significance 显著性水平liquid junction boundary 液接界面long range shielding effect 远程屏蔽效应magnetic equivalence 磁等价masking agent 掩蔽剂mass spectroscopy (MS) 质谱分析mass spectrum 质谱maximum absorption 最大吸收,吸收峰metabolite 代谢物metastable ion 亚稳离子micellar chromatography 胶束色谱法micelle 胶束microdialysis 微透析micropacked column 微型填充柱molarity 摩尔浓度monitor 检测;监控monochromator 单色器mortar 研钵multidimentional chromatography 多维色谱multiple linear regression 多元线性回归multiplet 多重(谱线)multivariate calibration 多元校正mass fragmentography 质量碎片图谱法micellar electrokinetic chromatography 胶束电动色谱microbore packed column 微径填充柱nonspecific impurity 一般杂质nonaqueous titration 非水滴定法normal distribution 正态分布normalization 归一化(法)nuclear magnetic resonance (NMR) 核磁共振far, mid, near infrared spectrum 远、中、近红外光谱octadecylsilane (ODS) 十八烷基硅烷octylsilane 辛烷基硅烷open tubular column 开管色谱柱optimal 最优的orthogonal function spectrophotometry 正交函数分光光度法ourlier 可疑数据;溢出值overtones 色彩oxygen flask combustion 氧瓶燃烧法odd electron 奇电子on-column injection 柱头进样packing material 色谱柱填充材料parent ion 母离子parts per million (ppm) 百万分之几peak width at half height 半峰宽pharmacopoeia 药典phosphorescence 磷光(photo) diode array detector 光电二极管阵列检测器polarimetry 旋光测定(法)polarography 极谱法porous-layer open tubular column 多孔层开管柱postcolumn 柱后potentiometer 电位计potentiometric titration 电位滴定precipitation form 沉淀形式precolumn 预柱pretreatment 预处理primary standard 基准物principal component regression 主成分回归法prototype drug 原型药物provisions for new drug approval 新药审批办法pseudophase chromatography 假相色谱法partition coefficient 分配系数phase boundary potential 相界电位platform balance 台秤;托盘天平precession 进动propagation of error 误差传递proton balance equation 质子平衡方程qualitative analysis 定性分析quantitative analysis 定量分析quadrupole mass spectrometer 四极质谱仪quasi-molecular ion 准分子离子radioimmunoassay 放射免疫分析法reagent sprayer 试剂喷洒器recovery 回收率reference electrode 参比电极refractive index 折射率repeatability 重复性reproducibility 重现性residue on ignition 炽灼残渣resolution 分辨率;分离度retention 保留reversed phase 反相rinse 清洗;淋洗robustness 可靠性;稳定性round 修约(数字)ruggedness 耐用性red shift 红移relative value (Rf) 相对比移值relative standard deviation (RSD) 相对标准偏差relaxation mechanism 弛豫历程saponification value 皂化值scatter diagram 散布图selectivity 选择性separating funnel 分液漏斗sieving 筛分signal to noise ratio 信噪比size exclusion chromatography (SEC) 空间排阻色谱法solid-phase extraction (SPE) 固相萃取solvent front 展开剂前沿sorbent 吸附剂specific absorptivity 比吸光系数specificity 专属性spectrophotometry 分光光度测定法spectrophotometer 分光光度计spectroscopic analysis 光谱分析spectroscopy 光谱法spectrum 光谱spiked 加入标准的split injection 分流进样spray reagent (平板色谱中的)显色剂standardization 标定stationary phase 固定相sterility test 无菌试验stirring bar 搅拌棒stock solution 原液stoichiometric method 化学计量方法stopcock 玻璃活塞stray light 漫射光supercritical fluid chromatography (SFC) 超临界流体色谱法support-coated open tubular column 涂层(担体)开管柱suspension 悬浊液swept field 扫场symmetry factor (fs) 对称因子system suitability (色谱)系统适应性systematic error 系统误差tailing factor 拖尾因子tailing suppressing reagent 扫尾剂therapeutic drug monitoring (TDM) 治疗药物监测thermal analysis 热分析thermal conductivity detector 热导检测器thermogravimetric analysis (TGA) 热重分析thermospray 热喷雾tight container 密闭容器time-resolved fluoroimmunoassay 时间分辨荧光免疫法titrant 滴定剂titer, titre 滴定度titrimetric analysis 滴定分析法tolerance 容许限transmittance 透光率turbidance 混浊turbidimetric assay 浊度测定法turbidity 浊度three-point chiral recognition model 三点手性识别模型ultraviolet 紫外线(的)uniformity of content/volume/weight 装量均匀性validation 确认;证实validity 可靠性variance 方差versus ——与——的关系曲线vicinal coupling 邻耦viscosity 粘度volatilization 挥发法voltammetry 伏安法;电流滴定法volumetric analysis 容量分析vortex mixer 旋涡混合器vibrational relaxation 振动弛豫volumetric precipitation method 容量沉淀法beaker 烧杯buret(te) 滴定管conical flask 锥形瓶crucible 坩埚cuvette, cell 比色杯;皿cylinder, graduate cylinder, measuring cylinder 量筒electrolytic cell 电解槽evaporating dish 蒸发皿flask 烧瓶funnel 漏斗measuring flask, volumetric flask 量瓶,容量瓶pipette 吸管platform balance 台秤transfer pipette 移液管wall-coated open tubular column 墙涂壁开管柱watch glass 表面皿weighing bottle 称量瓶weights 砝码well-closed container 封闭容器least square fitting 最小二乘法拟合partial least square method 偏最小二乘法weighed least square regression 加权最小二乘回归rocking, wagging, deformation, bending, scissoring, twisting vibration 面内摇摆,变形、弯曲、扭转振动singlet 单峰doublet(d) 双峰triplett 三重峰quartet 四重峰quintlet 五重峰sextet 六重峰multiplet 多重峰。
实验一 生化实验的基本知识、
(2)吸液 吸液 轻轻按下推动按钮, 轻轻按下推动按钮, 使按钮 由位置“ 推到位置 推到位置“ , 由位置“0”推到位置“1”,将 吸头垂直浸入液体2-4mm处, 处 吸头垂直浸入液体 缓慢松开按钮,即使按钮位置 缓慢松开按钮, 复位到“ 位 “1”复位到“0”位,完成吸液 复位到 程序, 程序,停1—2s后将吸头移出液 后将吸头移出液 若吸头表面残留液体, 面(若吸头表面残留液体,可用 若吸头表面残留液体 滤纸轻轻擦掉, 滤纸轻轻擦掉,注意勿触及吸
2、移液器及其使用方法 移液器构造
移液器(又称取液器、 移液器(又称取液器、 微量移液器) 枪、微量移液器)是一 种取液量连续可调的精 密取液仪器。 密取液仪器。 优点:重量轻、精度高、 优点:重量轻、精度高、 读数直观、使用方便。 读数直观、使用方便。
移液器的构造
移液器使用步骤
准备: 准备:
如图所示用右手握住移 液器。然后将移液器吸头 液器。 (枪头)套在移液器杆上,稍 枪头)套在移液器杆上, 加扭转压紧吸头使之与枪杆 间无空气间隙。 间无空气间隙。转动调节轮 至所需容积。 至所需容积。 移液器的正确持法
2.实验报告书写 2.实验报告书写
实验报告格式 实验报告应包括以下几方面: 实验报告应包括以下几方面: (1)实验题目 (2)实验目的 (3)实验原理 (4)实验主要材料 操作步骤(可用列表或流程图) (5)操作步骤(可用列表或流程图) 实验结果(记录、处理、得出结论)。 (6)实验结果(记录、处理、得出结论)。 讨论(分析) (7)讨论(分析)
朗伯-比尔定律 朗伯-
朗伯比尔定律的数学表达式 指一束平行单色光通 过一定液层厚度的有色溶 液时, 液时,溶质吸收了一部分 光能,光的强度(光的强 光能,光的强度( 度用“ 表示)就要减弱。 度用“I”表示)就要减弱。 溶液的浓度越大, 溶液的浓度越大,通过的 液层厚度越大, 液层厚度越大,则光被吸 收的越多,透过的越少。 收的越多,透过的越少。 表示入射光, (I0表示入射光,It表示 透过光) 透过光) 用T或A表示光透过的多少
吸光光度法
collimating Mirror
dispersion
prism (grating)
3. 吸收池 Absorption Cells (比色皿 Cuvettes) 材质:玻璃 / 石英 厚度:0.5/1/2/4cm
4. 检测器 Detector
光电管 Phototube
Cs3 Sb: 200~625nm AgOCs: 625~1000nm
第八章
吸光光度法( Absorptiometry)
分光光度法(Spectrophotometry)
§1
基本原理
一、光的波粒二象性
波动性:λ、υ、c υ= c /λ c = 3×108 m /s h = 6.63×10-34J· s 1nm = 10-9 m =10 Å
粒子性: E=hυ 1ev = 1.602×10-19 J
§ 2 分光光度计及吸收光谱
Spectrophotometer and absorption spectrum
一、分光光度计
光源
Source
单色器
吸收池
检测器
Detector
显 示 器
Indicator
Mono chromator Absorption Cell
1. 光源 (Light Source) 光源 钨灯 氢灯 氘灯 发射波长( n m ) 使用波长( n m ) 320~2500 150~400 350~1000 200~350
2. Lambert - Beer law A = lg I0 / It = - lgT = K b c
0 ~ 100% 0~∞
当一束 单色光 通过溶液时,溶液的吸光度与 溶液的浓度及液层厚度的乘积成正比。
荧光分光光度法
F= 2.303 I0cl 当入射光强度I0一定时,
F= Kc
(3-5) (3-6)
注意:上式在一定旳浓度范围内合用。
荧光物质旳最大浓度为c max 0.05/l。当浓度较 大时,即它旳吸光度不小于0.05时,荧光强度与其 浓
度旳线性关系将会发生偏离。
在浓度较高时,产生这种偏离旳原因可能是激
发分子间相互碰撞而失去能量(本身猝灭),或者
(7)荧光寿命 它是研究分子构造时要求旳参数。 定义:荧光强度衰减到1/e所需旳时间,用表达。
任意时间(t)旳荧光强度 If =If0e-t/=If0e-kt
式中:If —移去激发光源后任一时间t时旳荧光 强度;
If0—激发时最大旳荧光强度; k —仪器衰减常数; —激发态旳平均寿命。
(8)荧光分析旳敏捷度 —对整个发射光谱而言; /H —对部分发射光谱而言,即对所测到旳不
定其荧光值,然后将减去试剂空白荧光值旳原则 溶液荧光值与其相应浓度作图,即得其工作曲线。
根据试液及试液空白荧光值,在此曲线上即 可找到试液旳浓度。同步根据工作曲线旳线性情 况,能够拟定试液测定旳最高浓度。
c. 内标法
设Cx、Cs分别为试样和标样浓度,Fx、Fx0分 别为试样和试样本底旳荧光值;Fs+x为试样加标样 旳混合溶液旳荧光值。
基态跃迁到能级较高旳第一电子激发态或第二电子 激发态,然后经过无辐射跃迁返回到第一电子激发 态旳最低振动能级上,再从该能级降落至基态旳各 个不同旳振动能级上,同步释放出相应能量旳分子 荧光,最终以无辐射跃迁形式回到基态旳最低振动 能级。
需要注意旳是: (1)整个过程是在单线态之间进行旳;
(2)产生荧光旳过程极快,约在10-8 秒左右内完毕;
3.3 荧光分析旳措施及影响原因
甲醛法测定空气中二氧化硫影响因素分析
油气田环境保护ENVIRONMENTAL PROTECTION OF OIL&GAS FIELDS•48•2021年4月Vol.31No.2甲醛法测定空气中二氧化硫影响因素分析张亚文1陈卓2(1.中国石油大庆油田有限责任公司;2.北京林业大学)摘要甲醛吸收-副玫瑰苯胺分光光度法是测定环境空气中的二氧化硫的常用方法,该方法具有灵敏度高、选择性好等优点,但在实验分析过程中发现,该方法对实验室环境条件、反应条件、试剂的质量和实际操作要求严格。
文章通过对影响空白值及检测结果的显色温度、显色时间、实验试剂、样品的采集及保存等关键因素进行实验分析,使校准曲线斜率和试剂空白吸光度值在给定的检测条件下满足检测方法标准的要求,为提高甲醛法测定空气中二氧化硫测量结果的准确性提供参考。
关键词二氧化硫;甲醛法;影响因素DOI:10.3969/j.issn.1005-315&2021.02.011文章编号:1005-3158(2021)02-0048-03Analysis of Affecting Factors For Determination of SO2in AmbientAir by Formaldehyde MethodZhang Yawen1Chen Zhuo2(1.PetroChina Daqing Oilfield Co. ;2.Beijing Forestry University')ABSTRACT The formaldehyde absorption-pararosaniline spectrophotometry method is a common method for determination of sulfur dioxide in ambient air,this method has the advantages of high sensitivity and good selectivity.However,in the process of experimental analysis,it is found that this method need laboratory environmental conditions,reaction conditions,quality of reagents and actual operation requirements to be very strict.In this paper,the key factors that affect the blank value and detection results were analyzed, suchascolorationtemperature,durationtime,experimentalreagents,sampleco l ectionandpreservationetc Sothattheslopeofthecalibrationcurveandthereagentblankabsorbancevalue meetthegivendetection conditions.The requirements of the detection method standards provided a reference for improving the accuracyofthemeasurementresultsforsulfurdioxidedeterminationintheairbytheformaldehydemethod KEY WORDS sulphur dioxide;formaldehyde method;affecting factor0引言二氧化硫为无色有强烈刺激性气味的气体,它与大气中飘尘的协同作用会对哺乳动物的遗传信息和遗传物质造成损伤,同时也是形成酸雨的主要物质之一,它能够通过使土壤及水体的酸性发生改变而危害生态环境,同时二氧化硫也是衡量大气是否遭到污染的重要标志[1],随着人们的环保观念逐渐增强,二氧化硫已被列入国家重点控制的气态污染物[]。
分光光度分析法(精)
c0 空白 Blank
标 准 曲 线
A
c1
c2
c3
c4
标准溶液
Standard Sample
0.8
cx
待测溶液 Sample
0.6 Ax
0.4
0.2
0
cx
0
0.2
0.4
0.6
0.8
1
c (mmol/L)
5、 紫外—可见分光光度计
a、目视比色法 用眼睛比较溶液颜色的深浅以确定物质浓度的方
法称为目视比色法。 特点:设备简单、操作简便;无需单色光;准确度不 高。 b、分光光度计
日光:紫 蓝 青 绿 黄 橙 红
复合光:由各种单色光组成的光。如白光(太阳光)
单色光:只具有一种波长的光。 要求:=2nm。
互补色光:如果把两种适当颜色的光按一定的强度比 例混合也可以得到白光,这两种光就叫互补色光。
物质的颜色是由于物质对不同波长的光具有选择性的 吸收作用而产生的。如:CuSO4呈兰色。 物质呈现的颜色和吸收的光颜色之间是互补关系。
c. 样品室
样品室放置各种类型的吸收池( 比色皿)和相应的池架附件。 吸收池主要有石英池和玻璃池两 种。
d. 检测器
利用光电效应将透过吸收池的光信 号变成可测的电信号,常用的有光 电池、光电管或光电倍增管。
e. 结果显示记录系统
检流计、数字显示、微机进行仪 器自动控制和结果处理
6、 显色与测量条件的选择
500
550
600
650
波长(nm)
吸收曲线
吸收曲线清楚地描述了物质对光的吸收情况。
(1)不同物质吸收曲线的形状和吸收波长不同。
531
MnO4-
Spectrophotometry
SpectrophotometryDefinition: A quantitative measurement of light reflection or transmission properties of a material. Application:1. To identify unknown compounds by their characteristic absorption spectra.2. To determine the concentrations of analytes in solutions.SpectrophotometrySpectrophotometric measurement often use the absorption of light in the visible(between 380 and 760 nm) and ultraviolet regions ( 200- 380 nm ).Measurement of intensity of colored solutions (absorption of visible light) is also called Colorimetry.Part one:Applications of spectrophotometry – Identify unknown compoundsPrincipleThe extent to which a sample absorbs light depends strongly on the wavelength of light. (λ)For a new unknown material in a solution,measures the absorbance of the solution for each wavelength and construct the absorption spectrum by plotting A vsλ .Absorption Spectruma plot of absorbance vs wavelength and is characterized by the greatest wavelength (λmax)λmax* is characteristic of each compound and provides information on the electronic structure of a analyte.* obtain the highest sensitivity and to minimize deviations in quantitative measurementThe shape of the spectra , λmax (wavelength of maximal absorption) andε(molar extinction coefficient) can be used to identify the property of an unknown compoundsPart two:Applications of spectrophotometry —— determine the Concentration Unknown (quantitative measurement) Laws of Absorption of lightLambert-Beer’s Law:When a ray of monochromatic light passes through an uniform absorbing solution, Absorbance of the solution (also known as Optical Density or Extinction) is directly proportional to the concentration of the substance and the depth of the solution through which the light passes (cell path length).Equation for Lambert-Beer’s LawA=logI0/I= log1/T= K×C×LI0is the intensity of the incident lightI is the intensity of the transmitted lightK is absorption coefficient (the proportionality constant that depends on the absoring substance ,wavelength of light and the temperature)L is cell path lengthT is the transmittance of the lightε --Molar extinction coefficient:* when length ‘L’ is in 1 centimeter and concentration ‘C’ = 1mol/L , the absorbance is equal to ‘ε’* usually written as ε1mol/L and dimension of 1mol/L-1cm-1.Prerequisite of Lambert-Beer’s LawMonochromatic incident lightλmaxSolution is uniform and stablesolution concentration is not too high (A=0.2~0.7)How to determine Concentration Unknown?The assays are most sensitive at the extinction peak (absorption) of the substance.The absorption produced is due to the test substances (specific absorbance), not by the solvent and compounds in the reagents (nonspecific absorbance).Blank solution : contains all the fragments of standard and tests and undergoes the same stages but without the standard and test solution. This will help to exclude the absorption due to reagents.Standard solution :contains all the reagents of test and blank but it also includes a solution of known concentration of the same substance which is going to be determined in the test container.Test solution : it contains all the reagents as present in the blank and standard and undergoes the same steps, but it contains an unknown quantity of the substance.Two Methods1. Standard addition methodThe absorbances of the standard solution and test solution are measured .According to Lambert-Beer’s Law ,A S=K S*C S*L SA t =K t *C t *L tC t=A t/A s*C s2. Calibration curve methodCalibration curve (also called working curve)shows how A changes with the C of a solution.The curves are obtained by measuring the signal from a series of standards of known concentration.* The curves are used to determine the C unknown or to calibrate the linearity of an assay.•Prepare a series of standard solutions•Measure absorbances of the standard solutions•Prepare a calibration curve ( a plot of A vs C).•The absorbance of the unknown solution is used in conjunction with the calibration curve to determine the concentration of the analyte.Measurement of Light Absorption Components of Spectrophotometer* Source of Light* Monochromator – a device for selection of a band (wave-length) of light* Sample Container - Cuvette* Detector for unabsorbed radiant energy* Associated read out meters。
2.2.24. ABSORPTION SPECTROPHOTOMETRY, INFRARED
01/2008:20224 2.2.24. ABSORPTION SPECTROPHOTOMETRY, INFRAREDInfrared spectrophotometers are used for recording spectra in the region of 4000-650 cm−1 (2.5-15.4 µm) or in some cases down to 200 cm−1 (50 µm).APPARATUSSpectrophotometers for recording spectra consist of a suitable light source, monochromator or interferometer and detector.Fourier transform spectrophotometers use polychromatic radiation andcalculate the spectrum in the frequency domain from the original data byFourier transformation. Spectrophotometers fitted with an optical systemcapable of producing monochromatic radiation in the measurement region may also be used. Normally the spectrum is given as a function oftransmittance, the quotient of the intensity of the transmitted radiation and the incident radiation. It may also be given in absorbance.The absorbance (A) is defined as the logarithm to base 10 of the reciprocal of the transmittance (T):T =,I0 = intensity of incident radiation,I = intensity of transmitted radiation.PREPARATION OF THE SAMPLEFOR RECORDING BY TRANSMISSION OR ABSORPTIONPrepare the substance by one of the following methods.Liquids. Examine a liquid either in the form of a film between 2 platestransparent to infrared radiation, or in a cell of suitable path length, alsotransparent to infrared radiation.Liquids or solids in solution. Prepare a solution in a suitable solvent.Choose a concentration and a path length of the cell which give a satisfactory spectrum. Generally, good results are obtained with concentrations of 10-100 g/L for a path length of 0.5-0.1 mm. Absorption due to the solvent iscompensated by placing in the reference beam a similar cell containing the solvent used. If an FT-IR instrument is used, the absorption is compensated by recording the spectra for the solvent and the sample successively. The solvent absorbance, corrected by a compensation factor, is subtracted using calculation software.Solids. Examine solids dispersed in a suitable liquid (mull) or in a solid (halide disc), as appropriate. If prescribed in the monograph, make a film of a molten mass between 2 plates transparent to infrared radiation.A. MullTriturate a small quantity of the substance to be examined with the minimum quantity of liquid paraffin R or other suitable liquid; 5-10 mg of the substance to be examined is usually sufficient to make an adequate mull using one drop of liquid paraffin R. Compress the mull between 2 plates transparent toinfrared radiation.B. DiscTriturate 1-2 mg of the substance to be examined with 300-400 mg, unless otherwise specified, of finely powdered and dried potassium bromide R or potassium chloride R. These quantities are usually sufficient to give a disc of 10-15 mm diameter and a spectrum of suitable intensity. If the substance is a hydrochloride, it is recommended to use potassium chloride R. Carefully grind the mixture, spread it uniformly in a suitable die, and submit it to a pressure of about 800 MPa (8 t·cm−2). For substances that are unstable under normalatmospheric conditions or are hygroscopic, the disc is pressed in vacuo.Several factors may cause the formation of faulty discs, such as insufficient or excessive grinding, humidity or other impurities in the dispersion medium or an insufficient reduction of particle size. A disc is rejected if visual examination shows lack of uniform transparency or when transmittance at about 2000 cm−1(5 µm) in the absence of a specific absorption band is less than 60 per centwithout compensation, unless otherwise prescribed.Gases. Examine gases in a cell transparent to infrared radiation and having an optical path length of about 100 mm. Evacuate the cell and fill to thedesired pressure through a stopcock or needle valve using a suitable gastransfer line between the cell and the container of the gas to be examined.If necessary adjust the pressure in the cell to atmospheric pressure using a gas transparent to infrared radiation (for example nitrogen R and argon R). To avoid absorption interferences due to water, carbon dioxide or otheratmospheric gases, place in the reference beam, if possible, an identical cell that is either evacuated or filled with the gas transparent to infrared radiation.FOR RECORDING BY DIFFUSE REFLECTANCESolids. Triturate a mixture of the substance to be examined with finelypowdered and dried potassium bromide R or potassium chloride R. Use a mixture containing approximately 5 per cent of the substance, unlessotherwise specified. Grind the mixture, place it in a sample cup and examine the reflectance spectrum.The spectrum of the sample in absorbance mode may be obtained aftermathematical treatment of the spectra by the Kubelka-Munk function.FOR RECORDING BY ATTENUATED TOTAL REFLECTIONAttenuated total reflection (including multiple reflection) involves light being reflected internally by a transmitting medium, typically for a number ofreflections. However, several accessories exist where only one reflectionoccurs.Prepare the substance as follows. Place the substance to be examined in close contact with an internal reflection element (IRE) such as diamond,germanium, zinc selenide, thallium bromide-thallium iodide (KRS-5) oranother suitable material of high refractive index. Ensure close and uniform contact between the substance and the whole crystal surface of the internal reflection element, either by applying pressure or by dissolving the substance in an appropriate solvent, then covering the IRE with the obtained solution and evaporating to dryness. Examine the attenuated total reflectance (ATR) spectrum.IDENTIFICATION USING REFERENCE SUBSTANCESPrepare the substance to be examined and the reference substance by the same procedure and record the spectra between 4000-650 cm−1 (2.5-15.4 µm) under the same operational conditions. The transmission minima (absorptionmaxima) in the spectrum obtained with the substance to be examined correspond in position and relative size to those in the spectrum obtained with the reference substance (CRS).When the spectra recorded in the solid state show differences in the positions of the transmission minima (absorption maxima), treat the substance to be examined and the reference substance in the same manner so that they crystallise or are produced in the same form, or proceed as prescribed in the monograph, then record the spectra.IDENTIFICATION USING REFERENCE SPECTRAControl of resolution performance. For instruments having a monochromator, record the spectrum of a polystyrene film approximately35 µm in thickness. The difference x (see Figure 2.2.24.-1) between the percentage transmittance at the transmission maximum A at 2870 cm−1(3.48 µm) and that at the transmission minimum B at 2849.5 cm−1 (3.51 µm) must be greater than 18. The difference y between the percentage transmittance at the transmission maximum C at 1589 cm−1 (6.29 µm) and that at the transmission minimum D at 1583 cm−1 (6.32 µm) must be greaterthan 10.For Fourier-transform instruments, use suitable instrument resolution with the appropriate apodisation prescribed by the manufacturer. The resolution is checked by suitable means, for example by recording the spectrum of a polystyrene film approximately 35 µm in thickness. The difference between the absorbances at the absorption minimum at 2870 cm−1 and the absorption maximum at 2849.5 cm−1 is greater than 0.33. The difference between the absorbances at the absorption minimum at 1589 cm−1 and the absorption maximum at 1583 cm−1 is greater than 0.08.Verification of the wave-number scale. The wave-number scale may be verified using a polystyrene film, which has transmission minima (absorption maxima) at the wave numbers (in cm1) shown in Table 2.2.24.-1.Table 2.2.24.-1. Transmission minima and acceptable tolerances of a polystyrene film Transmissionminima (cm−1)Acceptable tolerance (cm−1)Monochromator instruments Fourier-transform instruments3060.0± 1.5± 1.02849.5± 2.0± 1.01942.9± 1.5± 1.01601.2± 1.0± 1.01583.0± 1.0± 1.01154.5± 1.0± 1.01028.3± 1.0± 1.0Method. Prepare the substance to be examined according to the instructions accompanying the reference spectrum/reference substance. Using the operating conditions that were used to obtain the reference spectrum, whichwill usually be the same as those for verifying the resolution performance, record the spectrum of the substance to be examined.The positions and the relative sizes of the bands in the spectrum of the substance to be examined and the reference spectrum are concordant in the 2 spectra.Compensation for water vapour and atmospheric carbon dioxide. For Fourier-transform instruments, spectral interference from water vapour and carbon dioxide is compensated using suitable algorithms according to the manufacturer s instructions. Alternatively, spectra can be acquired using suitable purged instruments or ensuring that sample and background single beam spectra are acquired under exactly the same conditions.Figure 2.2.24.-1. Typical spectrum of polystyrene used to verify theresolution performanceIMPURITIES IN GASESFor the analysis of impurities, use a cell transparent to infrared radiation and of suitable optical path length (for example, 1-20 m). Fill the cell as prescribed under Gases. For detection and quantification of the impurities, proceed as prescribed in the monograph.。
第3章紫外-可见分光光度法-精品文档
概述 一、定 义 二、常用的波长范围 三、定量依据 四、特 点 五、发展趋势
4
一、定 义
紫外-可见分光光度法:以紫外-可 见光为光源,照射被测物质,研 究其吸收光谱的分析方法。
• 属于电子光谱。
5
二、常用的波长范围
• 紫外光区:200~400nm • 可见光区:400~800nm
6
三、定量依据
• Lambert-Beer 定律: A=KLC
7
四、特 点
一)方法灵敏度较高
一般可测到 10-4~10-5g/ml 部分可达 10-7g/ml
二)测定准确度较高
一般为 0.5%~0.2%。
三)不能表现整个分子的特征
8
三)不能表现整个分子的特征
• 有机化合物的紫外吸收光谱主要反映 分子中发色团、助色团及其共轭的情 况,并不能表现整个分子的特征。
第三章 紫外可见分光光度法
1
概述 第1节 基本原理 第2节 Lambert-Ber定律 第3节 显色反应及其显色条件的选择 第4节 紫外-可见分光光度计 第5节 分析方法 第6节 应用与示例
2
• 主要内容 • 紫外-可见吸收光谱的基本概念 • 吸收光度法的基本原理 • 紫外-可见分光光度计 • 定量分析 • 定性分析 • 有机化合物分子结构研究简介 • 应用
15
一、紫外可见吸收光谱的产生
• 原子或分子中的电子总是处于某一种 运动状态之中,每一种状态都具有一 定的能量,属于一定的能级。
• 吸收外来幅射→被激发→从低能级跃 迁到较高能级(从基态跃迁到电子激 发态) → 电子光谱。
16
• 分子总能量: • E总=E内+E平+E振+E转+E电子 • 吸收辐射能: • △E= △E振+△E转+△E电子
傅里叶红外光谱仪测蛋白质
傅里叶红外光谱仪测蛋白质傅里叶红外光谱仪是一种常规的蛋白质分析工具,广泛应用于不同领域的研究中,如生物医学、生命科学和化学等。
该技术通过测量分子的红外吸收光谱来确定样品中的官能团。
在本文中,我们将详细介绍傅里叶红外光谱仪测量蛋白质的原理、方法、注意事项和数据分析。
一、原理红外光谱技术基于分子的振动吸收特性,是检测蛋白质构象和结构的重要手段之一。
蛋白质中的氨基酸残基的主链振动和侧链振动吸收红外辐射,进而反映出样品的官能团特征。
主链振动位于1650-1550cm^-1,侧链振动位于1550-600cm^-1。
通过测量这些振动能量的减少,可以确定蛋白质中的官能团类型和数量,进而推断出它的结构。
二、方法1. 样品制备蛋白质的样品制备对傅立叶红外光谱仪测量结果的准确性至关重要。
在进行测量前需要对样品的制备进行严格控制。
需要纯化和浓缩蛋白质样品。
将浓缩的样品溶解在合适的缓冲液并充分混合。
通过紫外吸收测定蛋白质的浓度,确保在红外光谱测量期间样品中的成分保持一致。
2. 样品测量在进行傅里叶红外光谱仪测量之前,需要将样品溶液置于红外吸收样品池并使其干燥。
然后使用红外光谱仪扫描吸收光谱范围(4000-400 cm^-1),并记录样品的红外光谱。
三、注意事项1. 液态样品需要建立基线;2. 液态和固态样品的取样方式、时间要求不同;3. 确保样品处于充分干燥状态,否则会影响热胀缩系数的测量精度。
四、数据分析傅里叶红外光谱仪得到的红外光谱是一个复杂的图谱,需要进行数据处理和分析才能得出有用的结论。
在进行数据分析前,首先需要建立一个有用的基线和峰度校正。
可以通过比较样品与标准样品的红外光谱,确定蛋白质样品中的官能团组成和数量。
通过结合其他分析手段(如X射线晶体学、NMR等)来推断蛋白质的构象和三维结构。
傅立叶红外光谱仪是一种非常有用的蛋白质分析工具,可以用于检测蛋白质样品中不同官能团的振动吸收特性。
通过合理的样品制备、测量方法和数据分析,可以得到有价值的蛋白质结构信息,进而推断蛋白质功能。
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a r X i v :a s t r o -p h /0505269v 1 12 M a y 2005Astronomy &Astrophysics manuscript no.2666February 2,2008(DOI:will be inserted by hand later)Absolute spectrophotometry of northern compactplanetary nebulae ⋆S.A.Wright 1,2,R.L.M.Corradi 1,and M.Perinotto 31Isaac Newton Group of Telescopes,Ap.de Correos 321,E-38700Sta.Cruz de la Palma,Canary Islands,Spain e-mail:rcorradi@ing.iac.es2UCLA,Department of Physics and Astronomy,Los Angeles,CA,90095,USA email:saw@3Dipartimento di Astronomia e Scienza dello Spazio,Universit´a di Firenze,L.go E.Fermi 5,I-50125Firenze,Italy,e-mail:mariop@arcetri.astro.itAbstract.We present medium-dispersion spectra and narrowband images of six northern compact planetary nebulae (PNe):BoBn 1,DdDm 1,IC 5117,M 1-5,M 1-71,and NGC 6833.From broad-slit spectra,total absolute fluxes and equivalent widths were measured for all observable emission lines.High signal-to-noise emission line fluxes of H α,H β,[O iii ],[N ii ],and HeI may serve as emission line flux standards for northern hemisphere observers.From narrow-slit spectra,we derive systemic radial velocities.For four PNe,available emission line fluxes were measured with sufficient signal-to-noise to probe the physical properties of their electron densities,temperatures,and chemical abundances.BoBn 1and DdDm 1,both type IV PNe,have an H βflux over three sigma away from previous measurements.We report the first abundance measurements of M 1-71.NGC 6833measured radial velocity and galactic coordinates suggest that it is associated with the outer arm or possibly the galactic halo,and its low abundance ([O/H]=1.3x10−4)may be indicative of low metallicity within that region.Key words.planetary nebulae:general –techniques:spec-troscopic:emission-line –flux:standards:chemical abun-dances1.IntroductionMeasuring absolute emission-line fluxes of planetary nebu-lae (PNe)is performed for a number of reasons.Primarily,absolute fluxes can be related to basic properties of the nebulae and their central stars (i.e.masses and luminosi-ties,once distances are independently known),and can be used to determine nebular physico-chemical properties.If total flux of a PNe is accurately measured,monitoring over decades can also reveal possible luminosity variations,which in turn are related to fast evolution of the central star during the PNe phase,especially for the largest core masses.In addition,absolute emission-line fluxes of PNe may also be used as standard calibrations for narrow band pho-2Wright et al.:Absolute spectrophotometry of planetarynebulaeFig.1.[N ii]images of four PNe discussed in this paper.North is at the top,East to the left.Thefield of view is10′′×10′′for the NOT images of BoBn1and M1-71,and5′′×5′′for the HST images of IC5117and NGC6833.Mathews1972,Barker1978).Recently,to improve on sim-ilar conventional techniques in the southern hemisphere,Dopita&Hua(1997)derived a set of southern emission-line standards by means of broad-slit absolute spectropho-tometry of a sample of compact PNe.Similar emission-linestandards are completely missing in the northern hemi-sphere.Along these lines,we have selected a small sampleof(supposedly)compact PNe observable from La Palma,and attempted to measure their integrated emission-linefluxes.To make full use of the wide spectral coverage,wehave measured absolutefluxes and equivalent widths forall emission lines present in our observations to investigatefurther the physical and chemical properties of these PNe.In Section2we explain the spectroscopic and narrowbandimaging observational details.Section3discusses the mor-phology and sizes of these PNe,and Section4presents theanalysis and results forfluxes,equivalent widths,radialvelocities and chemical abundances.Discussion and com-parison of our results to literature are made in Section5.2.Observations and reduction2.1.SpectroscopySpectroscopic observations were obtained at the 2.5mIsaac Newton Telescope(INT)of the Isaac Newton Groupof Telescopes,Roque de los Muchachos,La Palma,usingthe Intermediate Dispersion Spectrograph(IDS).Each ob-servation night was conducted under photometric condi-tions with seeing mostly between1and1.5arcsec,andonly rarely as high as2.5arcsec.Blue spectra,coveringthe range370-570nm,were obtained during three nightson May10,July11,and August12,2001with the EEV10CCD.Red spectra,covering the range520-710nm,wereobtained during two nights on August27and September4,2001with the Tek5CCD.Both red and blue portionswere measured with a632lines mm−1grating and235mmcamera yielding0.06nm pixel−1for the blue spectra and0.16nm pixel−1for the red spectra.For each object,a series of exposures were taken withincreasing exposure times from60seconds to600seconds,to achieve both bright and faint emission lines withoutsaturation.The list of objects observed may be found inTable1.Each exposure was observed with the spectrograph slitaligned to the parallactic angle to avoid loss of light dueto atmospheric differential refraction.We used a broad slitwith a width of7′′to ensure as much light as possible fromthe object passed through the slit.Further discussion ofslit loss may be found in Section4.We used a narrowerslit width of1.′′5for radial velocity measurements,andobserved CuNe arc lamps either before or after each ob-servation.IRAF software tasks in twodspec and image packageswere used as follows for data reduction.The CCD’s over-scan region was used for bias level subtraction in eachimage.For each night,a set of either domeflats or skyflats were used to remove pixel-to-pixel variation,and toeliminate cosmetic defects associated with the instrument.Row-by-row wavelength calibration was performed on thetwo-dimensional image,to account for any variations indispersion in the spatial direction.The task apall was used to define an extraction aper-ture large enough to include all of the emission-lines spa-tially,trace the object along the dispersions direction,sub-tract the background,and extract the object.Backgroundsubtraction was conducted at each wavelength from themedian of the background regions specified individuallyfor each observation.Flux calibration was carried out on all PNe with thegeneral set of spectrophotometric standards(Stone&Baldwin1983,Baldwin&Stone1984,Massey et al.1988,Oke1990).Standards were observed at varying times andairmass throughout the night to derive a nightly extinc-tion.The tasks standards,sensitivity,and calibrate,in or-der,were used for integrating the standard starfluxes overthe calibrated bandpasses,defining a sensitivity functionand extinction values,andflux calibrating each PNe.Spectra taken with the narrow slit were used to de-termine systematic radial velocities forfive of the tar-get PNe(BoBn1,DdDm1,IC5117,M1-71,and NGC6833).For M1-5,additional spectroscopic observations onSeptember24,2004were obtained at the4.2m WilliamHerschel Telescope with the ISIS red arm instrument.Aspectral range of607-713nm was measured using the500mm camera and1200lines mm−1,yielding a scale of0.23nm pixel−1.A slit width of0.′′5was used.Radial velocitesfor all PNe were computed using the average wavelengthWright et al.:Absolute spectrophotometry of planetary nebulae3 Table1.Observational details of target PNe.Diameters are computed.Object PN G R.A.(J2000)Dec Diameter V⊙Name Number[′′][km s−1]shift of spectral lines relative to their rest wavelengths, with the IRAF task rvidlines.2.2.ImagesCompactness of target PNe is a basic requirement to en-sure that the totalflux of the nebula is included in our broad slit spectroscopic observations.Target PNe were chosen to have a diameter smaller than4′′in Acker et al.(1992;hereafter A92).Sizes of the targets were further checked using narrowband images taken at the INT and 2.6Nordic Optical Telescope(NOT)of La Palma,and using HST archive images.Images from NOT were obtained on September9,19, 20,and October21,1997,using the ALFOSC instrument Loral2k×2k CCD,with a spatial scale of0.′′19per pixel. Narrowbandfilters were used to isolate the emission[N ii] at6583˚A from the nearby hydrogen Hα.For BoBn1,an [O iii]5007˚A image was also taken.Exposure times varied between30sec and10min,depending on the brightness of the nebula in each emission line and owing to the presence of thin clouds on some nights.The seeing varied between 0.′′7and1.′′0.A60sec image of M1-5was obtained with the prime-focus Wide Field Camera(WFC,pixel scale0′′.33)at the INT,on January29,2004.Thefilter used included both the Hαline and[NII]doublet.Seeing was1′′.4.Narrowband images of IC5117and NGC6833were retrieved from the HST public archive.They were taken with the PC CCD of the WFPC2camera for programmes n.6943and n.8307on1997and1999,respectively,using several narrowbandfilters including Hα,[N ii],and[O iii]. Exposure times range from3sec to10min.3.Morphology and sizesAs is common for PNe,the largest extension of the neb-ulae shows up in the low excitation line of[N ii],as com-pared to e.g.Hαand[O iii].The[N ii]images of four tar-gets are displayed in Figure1.IC5117and NGC6833 are well resolved by the HST.IC5117presents a highly elongated and clumpy morphology,possibly with a‘mul-tipolar’shape(cf.e.g.He2-47and M1-37,Sahai2000). NGC6833shows a bright nucleus from which two‘boxy’lobes depart.BoBn1and M1-71are also resolved in the ground-based images.BoBn1in the[N ii]image shows a small protrusion toward the South-East.M1-71is larger,and appears to be composed of a bright‘bulge’and a fainter disk-like structure extending approximately North-South.DdDm1and M1-5are only marginally resolved in our ground-based images,and no details of their morphology can be appreciated.For this reason,they are not displayed in Figure1.DdDm1does not show any sign of elonga-tion,while M1-5is slightly elliptical,as clearly shown in [O iii]and HβHST images kindly made available by Matt Bobrowsky.In Table1we list the sizes of the nebulae,measured as the diameter at the10%level of the peak surface bright-ness,deconvolved by the instrumental resolution profile (seeing in the case of ground-based images).This method was introduced by Tylenda et al.(2003)and proved to be a good diameter estimator for these barely resolved images. For M1-5,we used the HST image of Matt Bobrowsky.4.Analysis4.1.Linefluxes and equivalent widthsReduced spectra were transferred to the STARLINK spec-trum analysis program DIPSO(Howarth et al.1996)to measure emission-linefluxes and equivalent widths.Local continua werefit with afirst-degree polynomial on both sides of the emission-lines and subtracted from the spec-tra.Linefluxes were measured by integrating the contin-uum subtracted emission-lines between initial and ending points of emission features.Integration was performed to ensure we were deriving a‘measured’flux as opposed to a fittedflux(e.g.,from a Gaussianfit).Uncertainities were assigned to eachflux line measurement based upon that frame’s exposure time andflux calibration sensitivity.Equivalent widths were measured with respect to local linear continua.Errors for each equivalent width measure-ment were derived by DIPSO based upon scattering of linear continuumfit and signal-to-noise of each emission-line.Total absolutefluxes and equivalent widths listed in Tables3through8were computed from the weighted av-erage of individual measurements in the various spectra. Their respective errors,also listed in the Tables,were de-4Wright et al.:Absolute spectrophotometry of planetary nebulaerived from the weighted standard deviation of the individ-ual measurements for each emission line.It should be noted that most of the errors listed in Tables3-8are‘internal’errors which take into account the photon statistics of the observed PNe and standard stars, and the instrument and detector properties.In particular, this is the case when multiple observations for an object were all taken on a single night,and applies to the data of BoBn1(Table3),DdDm1(Table4),M1-5(Table6), and M1-71(Table7).When data are obtained in different nights,as for some emission lines of NGC6833and IC5117 (noted in Table8and Table5),a better estimate of‘ex-ternal’systematic errors is obtained,for instance from the use of particular standard stars,or a non-perfect knowl-edge of the atmospheric extinction.Errors for multiple nights are larger and should be considered more realistic than for observations based on just a single night.Another possible source of errors for the totalfluxes of our target PNe are slit losses due to thefinite width of the adopted slit.These have been kept as small as possible by selecting compact PNe and adopting a slit width of7′′for the spectroscopic observations.Slit losses have been esti-mated taking into account the size of each nebula in differ-ent emission lines from the present images,seeing effects, slit width and its orientation with respect to the long axis of the resolved nebula,as well as centering and guiding er-rors.Slit losses are small in the case of the smaller nebulae, namely BoBn1,DdDm1,and NGC6833.Considering that seeing during the observations was generally not worse than1.5arcsec,slit losses for these nebulae are estimated to be below0.5%.The effect is slightly higher for M1-5, were slit losses are estimated to be around1.0%.They are potentially larger for IC5117,especially in[N ii]where the nebula is most extended.However,except in M1-71,our slit loss was oriented along the major axis of its elongated morphology,and thus no light from the outer regions of the nebula has escaped;slit losses are estimated to be at the same level as thefirst three nebulae,namely below 0.5%.The only case in which slit losses are substantial (approximately3%in[N ii]and2%in Hα)is M1-71,as the slit(oriented along the parallactic angle)was cutting the nebula through its short axis,a fact only partially com-pensated by the fact that the emission is highly centrally concentrated.Including all the different sources of uncertainties dis-cussed above,our estimates of the errors for the total fluxes to be associated with each individual PN are pre-sented in Table2.They are obtained by summing quadrat-ically the errors due to slit losses with the average of the errors listed in Table3-8for all lines in each individual PN, or for only the brightest ones(Hβ,Hα,5007˚A,6583˚A, and6678˚A).The conclusion is that narrowband images in bright emission lines can be calibrated photometrically using all the nebulae in our sample except for M1-71to a level of1.4%or better,depending on the actual‘standard’PN adopted.At the same level,secular variations of totalflux Table 2.Estimated total errors for all emission lines fluxes,and for available bright lines,namely Hβ,Hα, 5007˚A,6583˚A,and6678˚A.Object Totlal error Total ErrorName(all lines)(bright lines)Bobn1 2.3% 1.1%DdDm1 3.8%<1%IC5117 2.6% 1.1%M1-5 2.3%<1%M1-71 3.1% 2.8%NGC6833 5.2%<1%Wright et al.:Absolute spectrophotometry of planetary nebulae5(hereafter;KB94).The low level of scatter in measure-ments for each emission-lineflux did propagate intofinal uncertainties in the derived abundances of the order of5 percent for helium and25percent for heavier elements.5.Discussion and ConclusionsThe spectra obtained were used to infer some physical and chemical properties of the target nebulae.This analysis generally confirms and extends results obtained by other authors.BoBn1and DdDm1are known to belong to the Galactic halo,owing to their high Galactic latitude,highly peculiar systemic radial velocity,and subsolar oxygen abundance(see also Barker&Cudworth1984,Howard et al.1997,and Dinerstein et al.2003).While ourflux ratios for DdDm1do not agree well with Dinerstein et al. 2003,we observe no systematic effects between the three line ratios that correspond to our wavelength range.An extensive spectroscopic study of IC5117was pre-sented by Hyung et al.(2001),who show that this nebula must be very young owing to its high gas densities,ranging from40000cm−3in the[Cl iii]and[S ii]emitting region up to100000cm−3from line diagnostics involving p3elec-trons.The large densityfluctuations found in this nebula are most likely related to the clumpy and highly asym-metrical morphology shown by HST images presented in this paper.We found rather high gas densities in M1-71. Parthasarathy et al.(1998)classified the central star of M1-71in the class of“weak emission line stars”(WELS) or[WC]-PG1159central stars.Our spectra confirm the presence of C IV580.6nm and465.0to468.6nm C IV-He II broad stellar features in emission,but not of any O V and O VI lines.To our knowledge,no abundance determinations are available for the nebula of M1-71prior to the present study.Our analysis confirms that,like in the other WELS and[WC]central stars of PNe,in spite of the fact that the central star of M1-71is helium and carbon rich,no pecu-liar gas abundances are found in the nebula compared to the general sample of Galactic PNe(cf.Corradi&Schwarz 1995).NGC6833has a peculiar systemic radial velocity,de-viating more than50km s−1from the general Galactic rotation,(even assuming a large distance of8-9kpc for this object).The galactic coordinates and radial velocity measurements of NGC6833coincide well with the outer arm structure observed in HI within the range of50.◦< l<195.◦,-5.◦<b<35.◦,and-175km s−1<V LSR<-60 km s−1(Wakker&van Woerden1991,Dwarakanath et al.2002).Low oxygen,neon and nitrogen abundances(cf. also the data for the global sample of PNe summmarized in Corradi&Schwarz1995),suggests that NGC6833is a nebula belonging to a relatively old stellar population. The electron density lower limit of10000cm3suggests that this nebula is likely to be in an early evolutionary stage.For NGC6833,our chemical abundances are in good agreement with those listed in Perinotto(1991). Abundances and T e([O iii])of IC5117,NGC6833,and DdDm1are also in good agreement with Mal’kov(1998), with abundances of He and heavier elements comparable within<5%,and effective temperatures deviating on av-erage by∼250K.By comparing our abundance measurements with KB94average abundance values for a particular class of PNe,we see that IC5117and M1-71fall within the range for non-type1PNe.Our abundance measurements for NGC6833(except He)are all significantly below the average KB94value for non-type1Pne;O,N,and Ne abundances are a factor of3.8,2.5,and3.6below,respec-tively and Ar and S are even further below.If NGC6833 is associated with the outer arm,then its abundance mea-surements would be an indicator of the outer arm’s metal-licity,which has been estimated to be lower than the inner disk(Digel er al.1990).However,we cannot exclude the possibility that it belongs to the Galactic halo.For four PNe,IC5117,M1-5,M1-71,and NGC6833, Hβflux falls within one sigma of A92reported Hβfluxes. BoBn1and DdDm1are greater than three sigma from A92.Hβmeasurements.This discrepancy between our measurements and A92may be due to underestimated uncertainities within the A92catalogue,which has been recently noted in Ruffle et al.2004.However,if theflux of BoBn1and DdDm1accurately represent intrinsic variability,then this calls for future monitoring of these sources.In conclusion,following the work done in the southern hemisphere by Dopita&Hua(1997),the present data provide an extension to the northern hemisphere of new emission-line standards for narrowband imaging.Further work of this kind is needed in order to have more complete coverage in right ascension and radial velocities. Acknowledgements.The authors thank Bego˜n a Garc´ıa for one night of service observations,Denise Gon¸c alves for further ob-servations,Danny Lennon for his thoughtful help in data re-duction and analysis,Remington P.S.Stone for valuable com-ments,and the referee for a number of helpful insights. 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