‘owz0is {8 Principle and Feature of Various Detection Methods (2): HitacN High- Technologies GLOBAL HITACHI Psecr @ corr spire the Nest Hitachi High-Technologies GLOBAL GlobalNetwork | Silmap Contr Us Products & Services AboutUS Suppor Semiconductor Warwactuing Equement Selence & Medical Systams Industnel IT Systons Advanced instal Products TOP Potite Sevens Scr KMesealSptis Tectia Supt Arabians Hot Pefeerance msi Chemtopep (HPLC) WLC eee Cause "Penrod Wet HPLC Basic Course 8. Principle and Feature of Various Detection Methods (2) ‘This chapter fs Fluorescence (FL) datecor’“Diferentialretractve index (RI) detector, "Conductivty detector "Elecvochomical detector (ECD),"Evaporatve ght scattering detector (ELSO)""Carona® Charged ‘Aerosol Detoctor® (Coronal CAD®)". Fluorescence (FL) detector [A.UVIUV.VIS detector monitors the absorption af light wth specified wavelength. However, some substances ‘absorb ight atone wavelength, and then emi ight called fhorescence at another wavelength. This is a jhenomenon in which a substance absorbs ight to reach a high-energy leveland then emits light o return to ts ‘original lave. Such a substance has specif wavelengths of ight that i absorbs (exctation wavelengths) and ‘emits (emission wavelengths). As familar examples, fliorescent paints and highlighters emvtfuorescence with @ ear color, Figure 1 shows @ FL detector optical system. While @ UVIUV-VIS detector detects ight that has passed through the flow cal, an FL detector detects uarescence emitted in the direction orthogonal to te exciting light. Bic Towel Bite a ean | atetier Ba TW suorascance Sapeienan , [Sage Xe lore DigacitY rane Fiqwe 1 Ciagemnmateustston at aFLestetor opal eyter (a etent a MACH Fluorescence detection is suitable for trace analysis because of generally having high sensivily and selectvty (not detecting impurtes). ‘There ave not mary components that orignal emit orescence (natural fuorescence). However, amino acd, ‘etc. can be detected as fuorescent substances, alter reaction with afuorescence ceagent (derivatization). This method makes it possible to measure various componenis wth high sensivy Differential refractive index (RI) detector [An Rl detector detects components based onthe refraction of ight in solution [As shown in Figure, the flow coll of an Rl detector is divided into the sample-sde and referance-side cel, Prior to analysis, eluate is introduced ito ether cel, ntl the ow of te eluate becomes equliorated, The reference Side cali filed wth eluate, and the column eluate i introduced into the sample-side cell through the changed flow channel, When components are eluted fram the column, the chemical composition changes inthe sample- ‘de solution, which changes ts photorefractive level [As ares, the amount of ight received by the ight-recelving section changes, showing a peak which can be detected. Any component inthe elite can be detected; thus, the RI detector is often called a “universal detector bpiwwwitachitightech,comiglobl orocits/sciencettechvanallctbasefcourse8 imi Anayicl Systoms ‘Thora Analysis High Perormance Lig (chromatograph (HPLC) Exams of Application by Cchromastor HPLC Basie Course Development of UHPLC: (ur Perermance-oentes ors ‘Spoctophotometer (UV Vis FL) ‘Atomic Avsrpton ‘Specrophetometer (AAS) ICP Analysis XRF Analysis Eloctron Microscope / Atomic Fores Microscope High Perfomance Liquid ‘Chromatogranh/ Amino Acc ‘Analyzer (HPLCINAA) 1“‘owz0is {8 Principle and Feature of Various Detection Methods (2): HitacN High- Technologies GLOBAL Lsheeouratame Lgetreconnd Figure? bhogrammaneiusvation of a Rlestactar opal eter Conduct ity detector Electric conductvty measurement of solution i a method of detecting ions inthe solution [tor the targted ions are olutod, the change in olectic current is detoctod, with a constant voltage imposad between the electrodes. A conductivity detector is employed as a detecor in an ion chromatograph, whichis a system dedicated to measuring ion. This detector is used mainly to measure inorganic fons and small organic ‘Substances, including organic acids and amines. “The ems shown below are often measured, Some ofthese items can be measured by atomic absorption ‘analysis. However, the ably to analyze negative lons simultaneously a particular mert of te on ‘chromatograph Nogative ions; FC, NOp., NO3-,Br-, S042-, P49. Positive ions; Nav; K+, NH +, Mg2*, Ca2* “The conductivity detector Is hghly sensitve, but very suscepti tothe effect of temperature variation (a change: (of Cin soktion temperature causes a change of roughly 2% in electric conductiviy). Various methods of ‘voiding temperature variations have been devised, such as constant-temperature cel, ‘Suppressor and non-suppressor Inthe ion etvomatagraph system, the electri conduciviy ofa solution is measured. A higher electric conductivity ofan eliate produces larger noise. Two measures fortis fact can be considered: An elvent is used that originally has low conductivity. + Non-suppressor method The elecic conductivity ofthe eluate ater column separation fs reduced, “+ Suppressor method ‘The suppressor method reduces the elactrc conductivity by replacing Na* with H* or replacing SO4?- with, (On the eluate through ion exchange resins or membranes. The nor-suppressor method fs easy and inexpensive, because a conductivity detector i simply added tothe typical LC system. Usual, the non suppressor method can bo used lo measure sampls with a concentration of roughly 0.1 ppm or more, Electrochemical detector (ECD) [An ECD is used to measure components displaying oxdationxeduction reactions, and detecs elactrc eurents ‘gonerated by these reactions. The ECD has high selectvly, because the necessary vollage o cause an ‘oxidaton-reducton reaction depends on the component. The ECD has also high sensi, and is oftan used for measurement of biogenic substances such as catecholamine. Evaporative light scattering detector (ELSD) [AN ELSD atomizes the column eluate, shines ight on the resulting particulate components, and detects the resulting scattered Ight. Theoretically, an ELSO can detect any nonvolatle component. An ELD has a sersitviy roughly 10 times higher than an Rl detector, but has a sightly low sensitivity to low molecular components due to their smal size, An ELSD is used mainly to detect non-UV-absorbing components. Attention shouls be given to ‘the fact that nonvolatile salts cannot be used as the eluent, Corona® Charged Aerosol Detector® (Corona® CAD®) Like an ELSD, a Corona CADE atomizes the column eluata to make the sample components particulate. However, a Corona® CAD® detects them electrically by ionizing them with charged Nz gas. A merit ofthe Corona® CAD isthe billy to detact components wth a sans higher than that of an ELSD, wth a sensitivity bpiwwwitachitightech,comiglobl orocits/sciencettechvanallctbasefcourse8 imi‘owz0is {8 Principle and Feature of Various Detection Methods (2): HitacN High- Technologies GLOBAL \Which depends only slighty on component. The eluent for a Corona® CAD® is similar to that for ELSD. ‘The mofis ofthe major detectors are summarized in Table 1 “Table 1. Mens of the vanous detectors Detector Selectvty Sensitivity Merits wane a ‘Seu to 19190 rm, Sry opus egy ot compe, ona Ai varity of subtances cane dated hat Den ttn Sry artrt@n0, 2 2 eros any ovine corpora, The seer ante cated er sh ord Conant iting resins canoe cosy Fomine g : Socks atta wih ih sry. Ts cnet elas Shon eater pesous ad pst cora civatton stoma Ay capone tha itr orate nom Sneed | 1 the oun can be cetactod, esp lw serait. Cube a prom geet aetr RN analysis. eviertve Ths dct alone clin ets, a igrecmeg , lect etre fh eng cae portale compote Nov ety es) Shyness winch, comaniviy 2 loicd congo dtc, Tis etre samt (2) (Seema fon ehoatoany Elctic creme etd ha ae goed lctechmih : by once ose eaten clos seat) Elsa xiv congeners costed in eect nonowret, tecon Coe Thich coun at and Se Cece cee eling paesite : 3 crews toed where Soca WV one ‘oraeating compe canbe decid wth aoe ‘erty Nob an tof ESD 4» Excl, 3 = Good, 2= Moser, 1 = Poor Rofer to Figure 3 to select detectors. Se es | [oer we Fo [Lv Ys DA Pl tiowssa=thesanneonaue Seer soci LEE [RESET] [Rene epmraestelaos| eure ampie} + ton anatysie sce nese oe Figur 3 alge or deen selector Fst Page Previews Page Pages ots HPLC Basie Course page top bpiwwwitachitightech,comiglobl orocits/sciencettechvanallctbasefcourse8 imi‘owz0is {8 Princiole and Feature of Various Detection Methods (2): HitacN High- Technologies GLOBAL | Prosucts & sonicas About Us Sericonter Manfecuing Easement ows & operate Protle contat Us Seance & Medal Sytem csr Grow Companies Indl 8 Syste Investor Relatons Advanced distil Proce | haem croup clbat site | asc sevnt ipsaton lina Can bpiwwwitachitightech,comiglobl orocits/sciencettechvanallctbasefcourse8 imi