HEALTH_Vol34No2_08

°“√æ—≤π“«‘∏’°“√µ√«®«‘‡§√“–Àå Biomarker ¢ÕߺŸâ —¡º— “√1,2-‰¥§≈Õ‚√Õ’‡∑𠉥§≈Õ‚√¡’‡∑π ·≈–‰µ√§≈Õ‚√‡Õ∏‘≈’πœ

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°“√æ—≤π“«‘∏’°“√µ√«®«‘‡§√“–Àå Biomarker ¢ÕߺŸâ —¡º— “√ 1, 2-‰¥§≈Õ‚√Õ’‡∑𠉥§≈Õ‚√¡’‡∑π ·≈– ‰µ√§≈Õ‚√‡Õ∏‘≈’π ¥â«¬‡∑§π‘§ Gas Chromatoghaphy-Electron Capture Detector-Headspace »‘√‘æ√ ‘ßÀå∑Õß π—°«‘∑¬“»“ µ√å°“√·æ∑¬å™”π“≠°“√ «‘°√¡ ®—π∑–‡π“«å π—°«‘∑¬“»“ µ√å°“√·æ∑¬åªØ‘∫—µ‘°“√ ™≈≥‘°“πµå “¬·°â« π—°«‘™“°“√ “∏“√≥ ÿ¢ ”π—°‚√§®“°°“√ª√–°Õ∫Õ“™’æ·≈– ‘Ëß·«¥≈âÕ¡ °√¡§«∫§ÿ¡‚√§

∫∑§—¥¬àÕ ªí®®ÿ∫—π‰¥â¡’°“√π” “√Õ‘π∑√’¬å‰Õ√–‡À¬ (Volatile Organic Compounds, VOCs) ¡“„™âª√–‚¬™πå„π™’«‘µ ª√–®”«—πÀ≈“¬Õ¬à“ß ‡™àπ ’∑“∫â“π πÈ”¬“øÕ° ’ πÈ”¬“´—°·Àâß πÈ”¬“¬âÕ¡º¡ “√¶à“·¡≈ß ·≈– µ—«∑”≈–≈“¬µà“ßÊ ‡ªìπµâπ “√ 1, 2-‰¥§≈Õ‚√Õ’‡∏𠉵√§≈Õ‚√‡Õ∏‘≈’π ·≈–‰¥§≈Õ‚√¡’‡∑𠇪ìπ “√∑’Ëæ∫¡“°∑’Ë ÿ¥µ‘¥Õ—π¥—∫ 1 „π 10 ¢Õß “√∑’Ë¡’§«“¡‡¢â¡¢âπ Ÿß„πÕ“°“»„π‡¢µπ‘§¡Õÿµ “À°√√¡¡“∫µ“æÿ¥ “√Õ‘π∑√’¬å‰Õ√–‡À¬‡À≈à“π’ÈÀ“°¡’°“√ – ¡‡ªì𠇫≈“π“π®–¡’º≈°√–∑∫µàÕ ‘Ëß·«¥≈âÕ¡ ‡ªìπÕ—πµ√“¬µàÕ ÿ¢¿“æ ·≈–Õ“®‡ªìπ “√°àÕ¡–‡√Á߉¥â ¥—ßπ—Èπ°“√µ√«®«‘‡§√“–Àå À“ biomarker ∑’ˇÀ¡“– ¡ ¢Õß°“√√—∫ —¡º— “√¥—ß°≈à“« ®÷߇ªìπ‡§√◊ËÕß¡◊Õ„π°“√«‘π‘®©—¬ ‡ΩÑ“√–«—ß ªÑÕß°—π‚√§·≈– ¿—¬∑’ˇ°‘¥®“°°“√√—∫ —¡º— “√æ‘… «—µ∂ÿª√– ߧå„π°“√»÷°…“π’È ‡æ◊ËÕæ—≤π“«‘∏’«‘‡§√“–Àå„À⇪ìπ«‘∏’¡“µ√∞“π ·≈– ∂à“¬∑Õ¥‡∑§‚π‚≈¬’∑’ˉ¥â ŸàÀπ૬ߓπ‡§√◊Õ¢à“¬ ‚¥¬„™â‡∑§π‘§ Gas chromatography «‘‡§√“–ÀåÀ“ 1, 2-‰¥§≈Õ ‚√Õ’‡∏π·≈–‰¥§≈Õ‚√¡’‡∑π‚¥¬µ√ß„πµ—«Õ¬à“߇≈◊Õ¥ à«π‰µ√§≈Õ‚√‡Õ∏‘≈’π ®–«‘‡§√“–ÀåÀ“ “√‡¡µ“∫Õ‰≈µå §◊Õ‰µ√ §≈Õ‚√Õ–´‘µ‘°·Õ´‘¥ ‚¥¬«‘∏’∑’ˉ¥âπ’ȉ¥âºà“π°√–∫«π°“√¬◊π¬—𧫓¡∂Ÿ°µâÕß (Method Validation) ‰¥â§à“ Limit of Detection (LOD) ·≈– Limit of Quantitation (LOQ) ¢Õß 1, 2-‰¥§≈Õ‚√Õ’‡∏𠉵√§≈Õ‚√Õ–´‘µ‘°·Õ´‘¥ ·≈– ‰¥§≈Õ‚√¡’‡∑π ‡∑à“°—∫ 0.1, 0.1, 0.01 mg/L ·≈– 1.0, 1.0, 0.1 mg/L µ“¡≈”¥—∫ ”À√—∫ %Recovery ¡’§à“‡∑à“°—∫ 102, 100 ·≈– 100 Precision ¡’§à“Õ¬Ÿà„π™à«ß Horwitz ûs Ratio (Typical HORRAT) §◊Õ‰¡à‡°‘π 1.5 ∑—Èß 3 √–¥—∫ §«“¡‡¢â¡¢âπ„π∑ÿ° “√ ‚¥¬¡’ Linearity ·≈– Range Õ¬Ÿà„π™à«ß 1-15 mg/L, 1-160 mg/L ·≈– 0.01-4 mg/L ·≈–¡’§à“ correlation coefficient ‡ªìπ 0.9996, 0.9997 ·≈– 0.9992 µ“¡≈”¥—∫ ®÷ß √ÿª‰¥â«à“ «‘∏’«‘‡§√“–Àå∑’ˉ¥â æ—≤π“¢÷Èπ‡æ◊ËÕ«‘‡§√“–ÀåÀ“ biomarker ¢Õß “√∑—Èß 3 ™π‘¥π’È “¡“√∂𔉪„™â‡ªìπ«‘∏’¡“µ√∞“π‰¥â

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Method development for the analysis of biomarker of 1,2-Dichloroethane, Trichloromethane and Trichloroethylene by Gas Chromatoghaphy-Electron Capture Detector-Headspace Siriporn Singthong Senior Medical Scientist Vigrom Janthanao Medical Scientist Chonnikarn Saikaew Public Health Officer Bureau of Occupational and Environmental diseases Department of diseases control

Abstract Nowadays, volatile organic compound (VOCs) are widely used as ingredients in household products such as paints, de-colouring agents, dry-cleaning solutions, cosmetics, insecticide and many other solvents. 1,2-Dichloroehane , Trichloroethylene and Dichloromethane are three of the ten VOCs air pollutants that found exceeded the air qualify standard in Map Ta Pud industrial area. These chemicals do not only have impact on the environment but also are harmful to human health. They have carcinogenic properties. Good analytical technique is important for detecting biomarkers of exposures, since the biomarkers are used for diseases diagnosis, diseases surveillance, and diseases protection. The objective of this study was to develop practical and reliable methods for analyzing the biomarkers of 1,2-Dichloroehane, Trichloroethylene and Dichloromethane. Determination of 1,2-Dichloroehane and Dichloromethane biomarkers in blood sample, and biomarker of Trichloroethylene, Trichloroacetic acid, acted in urine sample were performed. These analytical methods developed and validated using gas chromatography-electron capture-headspace (GC-ECD-HS) with limits of detection (LOD) and limits of quantification (LOQ) of 1,2 Dichloroethane, Trichloroacetic acid and dichloromethane were 0.1, 0.1, 0.01 mg/L and 1.0, 1.0, 0.1 mg/ L respectively. % Recovery of the three substance were 102, 100 and 100 respectively. Precision values in the range of Horwitz 's Ratio (Typical HORRAT) is not more than 1.5 and the concentrations of all three substances. The Linearity and Range of the three methods were in the range 1-15 mg/L, 1-160 mg/L and 0.01-4 mg/L and the correlation 0.9996, 0.9997 and 0.9992 respectively. It was concluded the analytical methods developed to analyze biomarker of the 3 of substances can be used as a standard method.

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°“√æ—≤π“«‘∏’°“√µ√«®«‘‡§√“–Àå Biomarker ¢ÕߺŸâ —¡º— “√1,2-‰¥§≈Õ‚√Õ’‡∑𠉥§≈Õ‚√¡’‡∑π ·≈–‰µ√§≈Õ‚√‡Õ∏‘≈’πœ

∫∑π” 1, 2-‰¥§≈Õ‚√Õ’‡∑𠉥§≈Õ‚√¡’‡∑π ·≈– ‰µ√§≈Õ‚√‡Õ∏‘≈’𠇪ìπÀπ÷Ëß„π “√Õ‘π∑√’¬å‰Õ√–‡À(Volatile Organic Compounds, VOCs) 20 ™π‘¥∑’˵âÕߥ”‡π‘π°“√‡ΩÑ“√–«—ß·≈–µ‘¥µ“¡µ√«® Õ∫„π‡¢µæ◊È π ∑’Ë π‘ § ¡Õÿ µ “À°√√¡¡“∫µ“æÿ ¥ ‡π◊ËÕß®“°æ∫«à“¡’°“√ªπ‡ªóôÕπ„π™—Èπ∫√√¬“°“»Õ¬Ÿà „π‡°≥±å∑’Ëπà“‡ªìπÀà«ß ·≈–‡ªìπ “√∑’Ë¡’Õ—πµ√“µàÕ¡πÿ…¬å·≈–¡’º≈°√–∑∫µàÕ ‘Ëß·«¥≈âÕ¡ ¥—ßπ—Èπ®÷ß ®”‡ªìπµâÕß¡’°“√æ—≤π“«‘∏’°“√µ√«®«‘‡§√“–Àå∑“ß Àâ Õ ßªØ‘ ∫— µ‘ ° “√‡æ◊Ë Õ À“µ— « ∫à ß ™’È ∑ “ß™’ « ¿“æ (biomarker) ¢Õß°“√√—∫ —¡º— “√Õ‘π∑√’¬å√–‡À‡À≈à“π’È ‡æ◊ËÕ„™â‡ªìπ‡§√◊ËÕß¡◊ÕÀπ÷Ëß„π°‘®°√√¡°“√ ªÑÕß°—𠧫∫§ÿ¡ ‚√§®“°°“√ª√–°Õ∫Õ“™’æ·≈– ‘Ëß·«¥≈âÕ¡ ‡π◊ËÕß®“°«‘∏’°“√µ√«®«‘‡§√“–Àå “√ µà“ßÊ ‡À≈à“π’Ȭ—߉¡à¡’«‘∏’¡“µ√∞“π (Standard method) „π°“√¥”‡π‘ π °√–∫«π°“√π’È ®÷ ß µâ Õ ß ‡√‘Ë¡°“√ ◊∫§âπ®“°ß“π«‘®—¬µà“ßÊ ‡æ◊ËÕÀ“«‘∏’°“√∑’Ë ‡À¡“– ¡ √«¡∑—È ß §«“¡‡ªì 𠉪‰¥â ‡æ◊Ë Õ „Àâ Õ¥§≈âÕß°—∫¿“«–‡»√…∞°‘® ‚¥¬„π°“√æ—≤π“«‘∏’ «‘‡§√“–Àå ¡’°“√¥—¥·ª≈ß·≈–ª√–¬ÿ°µå ¿“«–°“√ ∑”ß“π¢Õ߇§√◊Ë Õ ß °√–∫«π°“√∑¥ Õ∫∫“ß ª√–°“√„Àâ Õ¥§≈âÕß°—∫‡§√◊ËÕß¡◊Õ·≈–Õÿª°√≥å∑’Ë¡’ „πÀâ Õ ßªØ‘ ∫— µ‘ ° “√ ‚¥¬¬÷ ¥ À≈— ° §«“¡∂Ÿ ° µâ Õ ß ª√–À¬—¥ ·≈–‰¡à°Õà „À⇰‘¥¡≈æ‘…µàÕ ‘ßË ·«¥≈âÕ¡ «‘∏’ °“√∑’Ëæ—≤π“¢÷Èππ’È®–µâÕߺà“π°√–∫«π°“√¬◊π¬—𠧫“¡∂Ÿ°µâÕß (Method Validation) ®π‰¥â§à“∑’Ë ¡—Ëπ„®«à“«‘∏’∑’Ëæ—≤π“¢÷Èπ„À¡àπ’È “¡“√∂𔉪„™â‡ªìπ «‘∏’¡“µ√∞“π·≈– “¡“√∂∂à“¬∑Õ¥‡∑§‚π‚≈¬’∑’Ë æ—≤π“¢÷Èπ‰ª ŸàÀπ૬ߓπ∑’ˇ°’ˬ«¢âÕßµàÕ‰ª ‡∑§π‘§ Gas Chromatography-Electron Capture Detector Headspace ®“°°“√ ◊∫§âπß“π«‘®—¬µà“ßÊ ∑’˺à“π¡“ æ∫ «à“‡∑§π‘§ Gas Chromatography-Electron Cap-

ture Detection (GC-ECD) ‡ªì π ‡∑§π‘ § ∑’Ë ‡À¡“– ¡ ”À√—∫°“√µ√«®«‘‡§√“–Àå “√ª√–°Õ∫ Õ‘π∑√’¬å∑’Ë¡’§≈Õ√’π‡ªìπ à«πª√–°Õ∫ (organic chlorine compounds) „πµ—«Õ¬à“ß™’«¿“æ [1] ‚¥¡’À≈—°°“√§◊Õ‡¡◊ËÕ “√Õ‘π∑√’¬å‰Õ√–‡À¬„πµ—«Õ¬à“ß ™’«¿“扥â√—∫§«“¡√âÕπ∑’ˇÀ¡“– ¡®–∂Ÿ° àߺà“π ‡¢â“ Ÿà‡§√◊ËÕß GC ·≈–ºà“π‡¢â“ Ÿà§Õ≈—¡πå™π‘¥ high capacity fused silica capillary ∑”„À⇰‘¥°“√ ·¬°¢Õß “√ ‚¥¬¡’ carrier gas ‡ªìπµ—«æ“ “√ ‡§≈◊ËÕπ∑’˺à“π§Õ≈—¡πå ®“°π—Èπ “√∑’Ë∂Ÿ°·¬°®–ºà“π ‡¢â“ Ÿà‡§√◊ËÕßµ√«®«—¥™π‘¥ electron capture ‡æ◊ËÕ ∑”°“√µ√«®«—¥∑—Èß™π‘¥·≈–ª√‘¡“≥ “√ ®“°π—Èπ®÷ß ¡’°“√ª√–‡¡‘πº≈¥â«¬√–∫∫§Õ¡æ‘«‡µÕ√å °“√µ√«®«‘‡§√“–Àå Biomarker ¢ÕߺŸâ —¡º— “√ 1, 2-‰¥§≈Õ‚√Õ’‡∑π ¡’ √ “¬ß“π°“√»÷ ° …“∑’Ë ºà “ π¡“∂÷ ß ‡∑§π‘ § °“√µ√«®«‘ ‡ §√“–Àå 1, 2-‰¥§≈Õ‚√Õ’ ‡ ∑π„π ¢Õ߇À≈«·≈–‡π◊ÈÕ‡¬◊ËÕµà“ßÊ ¢Õß√à“ß°“¬ ¥â«‡∑§π‘§ Gas chromatography/mass spectrophotometry (GC/MS) ‚¥¬∑—Ë « ‰ª¡— ° „™â «‘ ∏’ °“√µ√«®«—¥ 1, 2-‰¥§≈Õ‚√Õ’‡∑π®“°µ—«Õ¬à“ß Õ“°“» ‡≈◊Õ¥·≈–ªí “«– [2,3,4,5] ´÷Ë߇ªìπ‡∑§π‘§∑’Ë ¡’§«“¡‰« Ÿß¡“° “¡“√∂µ√«®«—¥‰¥â„π™à«ß√–¥—∫ µË”°«à“ ppb à«πÕ’°«‘∏’‡ªìπ°“√µ√«®«—¥‚¥¬«—¥ ®“°°‘®°√√¡°“√∑”ß“π¢Õ߇ÕÁπ‰´¡å GlutathioneS-transferase (GST) ‡æ◊ËÕµ√«®À“ 1, 2-‰¥ §≈Õ‚√Õ’‡∑π„π‡¡Á¥‡≈◊Õ¥·¥ß ·µà‡π◊ËÕß®“°º≈∑’ˉ¥â ¡’§«“¡§≈⓬§≈÷ß°—∫ “√Õ◊Ëπ §◊ÕÕ–‚§√‡≈Õ’π ‚æ√ ‰æ√’πÕÕ°‰´¥å ‰µ√’πÕÕ°‰´¥å ·≈–‡Õ∏‘≈’π‰¥ ‚∫√‰¡¥å ®÷߇ªìπ‰ª‰¡à‰¥â∑’Ë®–„™â GST ‡ªìπµ—«™’È«—¥ ∑“ß™’«¿“æ [6] πÕ°®“°π’Ȭ—߉¡à¡’«‘∏’°“√µ√«® «‘‡§√“–À凡µ“∫Õ‰≈µå¢Õß 1, 2-‰¥§≈Õ‚√Õ’‡∑π „πªí “«–‡ªìπß“πª√–®” ·¡â«à“®–¡’°“√·π–π” „Àâ„™â 2-§≈Õ‚√‡Õ∏“πÕ≈·≈–‚¡‚π§≈Õ‚√Õ–´‘µ‘°

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∫ ∑ «‘ ∑ ¬ “ ° “ √ ·Õ´‘¥„πªí “«–‡ªìπµ—«™’È«—¥∑“ß™’«¿“æ∑’Ë„™â‡ªìπ À≈—°∞“π· ¥ß„Àâ‡ÀÁπ«à“¡’°“√ —¡º— “√„π°≈ÿà¡ ¢Õߧ≈Õ√‘‡πµ‰Œ‚¥√§“√å∫Õπ [7] ·µà‡π◊ËÕß®“° “√ ‡¡µ“∫Õ‰≈µå ∑—È ß Õß™π‘ ¥ π’È ‰ ¡à „ ™à “√∑’Ë ¡’ § «“¡ ®”‡æ“–µàÕ 1, 2-‰¥§≈Õ‚√Õ’‡∑π‡æ’¬ßÕ¬à“߇¥’¬« ∑”„Àâ‰¡à “¡“√∂„™â “√‡¡µ“∫Õ‰≈µå∑—Èß Õß™π‘¥π’È ‡ªìπµ—«™’È«—¥∑“ß™’«¿“扥⠥—ßπ—Èπ®÷ß„™â«‘∏’°“√ µ√«®À“ 1, 2-‰¥§≈Õ‚√Õ’‡∑π‚¥¬µ√ß®“°°“√ À“¬„®ÕÕ° ‡≈◊Õ¥·≈–ªí “«–·∑π πÕ°®“°π’È °“√µ√«®«‘ ‡ §√“–Àå 1, 2-‰¥§≈Õ‚√Õ’ ‡ ∑π„π µ—«Õ¬à“ßÕ“°“»¡—°π‘¬¡„™â‡∑§π‘§ GC/MS ·≈– GC/ECD [8,9,10] ´÷Ëß¡’§«“¡‰«Õ¬Ÿà„π™à«ß sub-ppb ∂÷ß low-ppt ¡’ % Recovery ¡“°°«à“ 90 % ·≈– ¡’§«“¡·¡à𬔠Ÿß (3% RSD) [11,12] °“√µ√«®«‘‡§√“–Àå Biomarker ¢ÕߺŸâ —¡º— “√‰¥§≈Õ‚√¡’‡∑π °“√À“µ— « ™’È «— ¥ ∑“ß™’ « ¿“æ¢Õß°“√√— ∫ — ¡ º— “√‰¥§≈Õ‚√¡’ ‡ ∑π„π√à “ ß°“¬ “¡“√∂ µ√«®À“‰¥â À ≈“¬∑“ߧ◊ Õ ®“°‡≈◊ Õ ¥ [13] ªí “«– ≈¡À“¬„® [14] ‡π◊ÈÕ‡¬◊ËÕ‰¢¡—π [15] ·≈–πÈ”π¡ [16] ‚¥‡∑§π‘§∑’πË ¬‘ ¡„™â§Õ◊ ‡∑§π‘§ Gas chromatography/ Flame Ionization Detector (GC/FID) ·≈– GC/MS ´÷Ë߇ªìπ«‘∏’∑’Ë¡’§«“¡‰«æÕ∑’Ë®– “¡“√∂ µ√«®À“‰¥§≈Õ‚√¡’‡∑π„π√à“ß°“¬‰¥â ·µàÕ¬à“߉√ °Á µ “¡°Á ¬— ß µâ Õ ß¡’ ° “√À“§à “ µË” ÿ ¥ ∑’Ë ‡ §√◊Ë Õ ß¡◊ Õ “¡“√∂«‘‡§√“–À剥Ⱇ∫§«“¡∂Ÿ°µâÕߢÕß«‘∏’ „π °√≥’∑’Ë¡’°“√√—∫ —¡º— “√„π√–¥—∫µË” °“√µ√«®«‘‡§√“–Àå Biomarker ¢ÕߺŸâ —¡º— “√‰µ√§≈Õ‚√‡Õ∏‘≈’π °“√µ√«®«‘‡§√“–ÀåÀ“µ—«™’È«—¥∑“ß™’«¿“æ ¢Õß°“√√—∫ —¡º— ‰µ√§≈Õ‚√‡Õ∏‘≈’π„π√à“ß°“¬¡’ À≈“¬«‘∏’ «‘∏’À≈—°∑’Ë„™â«‘‡§√“–ÀåÀ“ “√‡¡µ“∫Õ‰≈µå ¢Õ߉µ√§≈Õ‚√‡Õ∏‘≈’π§◊Õ®–«‘‡§√“–ÀåÀ“‰µ√§≈Õ‚√ ‡Õ∏“πÕ≈·≈–‰µ√§≈Õ‚√Õ–´’µ°‘ ·Õ´‘¥ (TCA) ‚¥86

‡¡…“¬π - ¡‘∂ÿπ“¬π 2554

°√¡Õπ“¡—¬ à߇ √‘¡„Àâ§π‰∑¬ ÿ¢¿“楒

„™â‡∑§π‘§ GC/MS À√◊Õ GC/ECD ´÷Ëß “¡“√∂ µ√«®À“‰¥â∑—Èß„π≈¡À“¬„® ‡≈◊Õ¥ ªí “«– πÈ”π¡ ·≈–„π‡π◊ÈÕ‡¬◊ËÕµà“ßÊ [17,18,19,20,21] °“√«‘‡§√“–Àå‚¥„™â headspace ®–¥Ÿ¥™—Èπ°ä“´∑’ËÕ¬Ÿà‡Àπ◊Õµ—«Õ¬à“ß ‡¢â “ Ÿà ‡ §√◊Ë Õ ß GC °“√«‘ ‡ §√“–Àå À “‰µ√§≈Õ‚√ ‡Õ∏“πÕ≈ ·≈– TCA π—Èπ ®–µâÕß∑”‚¥¬„™â‡∑§π‘§ headspace GC/ECD °àÕπ ‡æ◊ËÕ‡ª≈’ˬπ TCA „Àâ ‡ªì π §≈Õ‚√øÕ√å ¡ ®“°π—È π ‡§√◊Ë Õ ß®–∑”°“√ «‘‡§√“–Àå “√„π√Ÿª¢Õ߉µ√§≈Õ‚√‡Õ∏“πÕ≈·≈– §≈Õ‚√øÕ√å¡ ´÷Ëß«‘∏’π’È¡’§«“¡∂Ÿ°µâÕß·≈–·¡à𬔠Ÿß “¡“√∂µ√«®«‘‡§√“–À剥â∂÷ß√–¥—∫ ppb

«—µ∂ÿª√– ߧ尓√»÷°…“ ‡æ◊Ë Õ æ— ≤ π“«‘ ∏’ ° “√µ√«®«‘ ‡ §√“–Àå “√ Õ‘π∑√’¬å√–‡À¬„πµ—«Õ¬à“ß™’«¿“æ∑’ˇªìπ¡“µ√∞“π ·≈–π”«‘ ∏’ ° “√µ√«®«‘ ‡ §√“–Àå ∑’Ë ‰ ¥â æ— ≤ π“¢÷È π π—∫ πÿπ°“√µ√«®«‘‡§√“–Àå·≈–∂à“¬∑Õ¥„ÀâÀâÕß ªØ‘∫—µ‘°“√∑’ˇªìπ‡§√◊Õ¢à“¬π”‰ª„™â„π°“√¥”‡π‘π ß“πªÑÕß°—𠧫∫§ÿ¡‚√§®“°°“√ª√–°Õ∫Õ“™’æ ·≈– ‘Ëß·«¥≈âÕ¡

«‘∏’°“√»÷°…“ 1. °“√‡µ√’ ¬ ¡ “√≈–≈“¬¡“µ√∞“π (Standard preparation) °“√‡µ√’¬¡ “√≈–≈“¬¡“µ√∞“π¢Õß 1, 2‰¥§≈Õ‚√Õ’‡∑π ‚¥¬≈–≈“¬ 1, 2-‰¥§≈Õ‚√Õ’‡∑π „π‡¡∑“πÕ≈„Àâ¡’§«“¡‡¢â¡¢âπ 10 g/L ·≈–π” “√≈–≈“¬∑’ˉ¥â¡“‡®◊Õ®“ߥ⫬πÈ”°≈—Ëπ„Àâ¡’§«“¡ ‡¢â ¡ ¢â π 50 mg/L ®“°π—È π π”¡“‡µ√’ ¬ ¡‡ªì π “√≈–≈“¬¡“µ√∞“π„π‡≈◊Õ¥∑’Ë¡’§«“¡‡¢â¡¢âπ 1, 1.5, 2, 2.5, 5, 10, 15 ·≈– 17.5 mg/L °“√‡µ√’¬¡ “√≈–≈“¬¡“µ√∞“π¢Õ߉¥§≈Õ ‚√¡’‡∑π ‚¥¬≈–≈“¬‰¥§≈Õ‚√¡’‡∑π„π‡¡∑“πÕ≈ „Àâ¡’§«“¡‡¢â¡¢âπ 10 g/L ·≈–π” “√≈–≈“¬∑’ˉ¥â

°“√æ—≤π“«‘∏’°“√µ√«®«‘‡§√“–Àå Biomarker ¢ÕߺŸâ —¡º— “√1,2-‰¥§≈Õ‚√Õ’‡∑𠉥§≈Õ‚√¡’‡∑π ·≈–‰µ√§≈Õ‚√‡Õ∏‘≈’πœ

¡“‡®◊Õ®“ߥ⫬πÈ”°≈—πË „À⡧’ «“¡‡¢â¡¢âπ 10 mg/L ®“°π—Èππ”¡“‡µ√’¬¡‡ªìπ “√≈–≈“¬¡“µ√∞“π„π ‡≈◊Õ¥∑’Ë¡’§«“¡‡¢â¡¢âπ 0.01, 0.1, 1, 2, 3 ·≈– 4 mg/L °“√‡µ√’ ¬ ¡ “√≈–≈“¬¡“µ√∞“π¢Õ߉µ√ §≈Õ‚√Õ–´‘µ‘°·Õ´‘¥ ‚¥¬≈–≈“¬‰µ√§≈Õ‚√Õ–´‘ µ‘°·Õ´‘¥„ππÈ”°≈—Ëπ„Àâ¡’§«“¡‡¢â¡¢âπ 10 g/L ·≈– π” “√≈–≈“¬∑’Ë ‰ ¥â ¡ “‡®◊ Õ ®“ߥ⠫ ¬ªí “«–„Àâ ¡’ §«“¡‡¢â¡¢âπ 1 g/L ®“°π—Èππ”¡“‡µ√’¬¡‡ªìπ “√≈–≈“¬¡“µ√∞“π„π‡≈◊Õ¥∑’¡Ë §’ «“¡‡¢â¡¢âπ 1, 5, 10, 20, 40, 80 ·≈– 160 mg/L 2. °“√‡°Á∫·≈–‡µ√’¬¡µ—«Õ¬à“ß (Sample preparation) µ—«Õ¬à“߇≈◊Õ¥ ‡°Á ∫ µ— « Õ¬à “ ߇≈◊ Õ ¥ 5 mL „ à „ π vaccutainner tube ∑’¡Ë °’ “√‡§≈◊Õ∫º‘«¿“¬„πÀ≈Õ¥ ¥â«¬ “√°—π‡≈◊Õ¥·¢Áß ‚¥¬¡’™àÕß«à“ß¿“¬„πÀ≈Õ¥ ‡À≈◊ÕÕ¬ŸàπâÕ¬∑’Ë ÿ¥ ·™à‡¬Áπ (4 ÌC) ∑—π∑’°àÕππ” àß À“°®”‡ªìπµâÕ߇°Á∫„πµŸâ‡¬Áππ“π°àÕπ àßµ√«® „Àâ ‡°Á∫‡¢â“™àÕß·¢Áß °“√‡µ√’¬¡µ—«Õ¬à“߇≈◊Õ¥ ”À√—∫«‘‡§√“–Àå ‚¥¬ ªî‡ªµ‡≈◊Õ¥®”π«π 1 mL „ à≈ß„π¢«¥ ”À√—∫ Headspace-Sampler ®“°π—Èπªî¥Ω“¥â«¬®ÿ°¬“ß ´‘≈‘‚§π·≈–Ω“Õ≈Ÿ¡‘‡π’¬¡ ∫’∫„Àâ·πàπ¥â«¬ crimper 𔉪‡¢â “ ‡§√◊Ë Õ ß Headspace-Sampler ‡æ◊Ë Õ ∑”°“√«‘‡§√“–Àå¥â«¬‡∑§π‘§ GC-HS µ—«Õ¬à“ßªí “«– ‡°Á∫µ—«Õ¬à“ßªí “«– 10 mL À“°¬—߉¡à ∑”°“√«‘‡§√“–Àå„π∑—π∑’„À⇰Á∫√—°…“∑’ËÕÿ≥À¿Ÿ¡‘ 4 ÌC “¡“√∂‡°Á∫‰¥âπ“πª√–¡“≥ 1 —ª¥“Àå À“° ‡°Á∫∑’ÕË ≥ ÿ À¿Ÿ¡‘ -20 CÌ “¡“√∂‡°Á∫‰¥âπ“πª√–¡“≥ 2 ‡¥◊Õπ °“√‡µ√’¬¡µ—«Õ¬à“ßªí “«– ”À√—∫°“√«‘ ‡§√“–Àå‚¥¬ªî‡ªµªí “«– 1 mL „ à≈ß„π¢«¥

”À√—∫ Headspace-Sampler ®“°π—Èπªî¥Ω“¥â«®ÿ°¬“ß´‘≈‘‚§π·≈–Ω“Õ≈Ÿ¡‘‡π’¬¡ ∫’∫„Àâ·πàπ¥â«crimper 𔉪‡¢â“‡§√◊ËÕß Headspace-Sampler ‡æ◊ËÕ∑”°“√«‘‡§√“–Àå¥â«¬‡∑§π‘§ GC-HS 3. °“√«‘‡§√“–À奫⠬‡∑§π‘§ GC-ECDHS (GC-ECD-HS analysis) °“√µ√«®«‘‡§√“–ÀåÀ“ Biomarker ¢ÕߺŸâ —¡º— “√ 1, 2-‰¥§≈Õ‚√Õ’‡∑𠉵√§≈Õ‚√Õ–´‘ µ‘°·Õ´‘¥·≈–‰¥§≈Õ‚√¡’‡∑𠇧√◊ËÕß¡◊Õ∑’Ë„™â„π°“√ «‘‡§√“–Àå§◊Õ Gas Chromatography (GC) ∑’Ë “¡“√∂«‘‡§√“–Àå¥â«¬«‘∏’ Headspace §Õ≈—¡πå∑’Ë „™â·¬° “√‡ªìπ§Õ≈—¡πå HP-FFAP Polyethylene Glycol TPA, 0.32 mm x 50 m ·≈–„™â Œ’‡≈’¬¡‡ªìπµ—«π”°ä“´ (1.5 mL/min) „π°“√©’¥ µ—«Õ¬à“ß„™â Split mode ∑’ËÕÿ≥À¿Ÿ¡‘ 250 ÌC ‚¥µ—Èß‚ª√·°√¡ column oven ®“° 80 ÌC ∂÷ß 120 ÌC (10 ÌC/min) ·≈–®“° 120 ÌC ∂÷ß 150 ÌC (10 ÌC/ min) „™â‡«≈“ 9 π“∑’ ”À√—∫°“√«‘‡§√“–ÀåÀ“ 1,2‰¥§≈Õ‚√Õ’‡∑π·≈–‰¥§≈Õ‚√¡’‡∑π„π‡≈◊Õ¥ ·≈–„™â ‚ª√·°√¡ column oven 80 ÌC ‡ªìπ‡«≈“ 12 π“∑’ ”À√—∫°“√«‘‡§√“–À創√§≈Õ‚√Õ–´‘µ‘°·Õ´‘¥„π ªí “«– 4. °“√¬◊ π ¬— 𠧫“¡∂Ÿ ° µâ Õ ß¢Õß«‘ ∏’ (Validation of the analytical method) - °“√∑¥ Õ∫À“§à“µË” ÿ¥¢Õß«‘∏«’ ‡‘ §√“–Àå (Limit of detection, LOD) ·≈–§à“µË” ÿ¥∑’Ë ¬Õ¡√—∫‰¥â¢Õß«‘∏«’ ‡‘ §√“–Àå (Limit of quantitation, LOQ) °“√À“§à“ LOD ·≈– LOQ ‚¥¬‡µ√’¬¡ sample blank ·¬°°—π 10 µ—«Õ¬à“ß ®“°π—Èπ®÷ß «‘‡§√“–Àå¥â«¬‡∑§π‘§ GC-ECD-HS ∑’Ëæ—≤π“¢÷Èπ ·≈–§”π«≥À“§à“‡∫’ˬ߇∫π¡“µ√∞“π (SD) ‚¥¬ LOD = Mean ± 3SD LOQ = Mean ± 10SD

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∫ ∑ «‘ ∑ ¬ “ ° “ √ - °“√∑¥ Õ∫À“§à “ °“√∑”´È” Õ¬à “ ß ·¡à𬔷≈–§«“¡≈”‡Õ’¬ß¢Õß«‘∏’«‘‡§√“–Àå (Precision and Bias) ∑¥ Õ∫‚¥¬«‘ ‡ §√“–Àå µ— « Õ¬à “ ߇≈◊ Õ ¥À√◊ Õ ªí “«–∑’Ë ‡ µ‘ ¡ “√¡“µ√∞“π·µà ≈ –™π‘ ¥ ≈߉ª∑’Ë §«“¡‡¢â¡¢âπ 3 √–¥—∫ (µË” °≈“ß Ÿß) ¥—ßπ’È µ—«Õ¬à“ß ‡≈◊Õ¥∑’ˇµ‘¡ “√¡“µ√∞“π 1,2-‰¥§≈Õ‚√Õ’‡∏π∑’Ë §«“¡‡¢â¡¢âπ 1, 10 ·≈– 15 mg/L µ—«Õ¬à“ß ‡≈◊Õ¥∑’ˇµ‘¡ “√¡“µ√∞“π‰¥§≈Õ‚√¡’‡∏π∑’˧«“¡ ‡¢â ¡ ¢â π 0.1, 1 ·≈– 2 mg/L ·≈–µ— « Õ¬à “ ß ªí “«–∑’ˇµ‘¡ “√¡“µ√∞“π‰µ√§≈Õ‚√Õ–´’µ‘°·Õ ´‘¥∑’˧«“¡‡¢â¡¢âπ 1, 80 ·≈– 160 mg/L ®“° π—Èπ®÷ßπ”¡“«‘‡§√“–À姫“¡‡¢â¡¢âπ≈– 10 ´È” - °“√∑¥ Õ∫À“§«“¡∂Ÿ ° µâ Õ ß¢Õߺ≈ °“√∑¥ Õ∫

°“√µ√«®«‘‡§√“–À嵫— Õ¬à“ß∑’‰Ë ¡à∑√“∫§à“ (Analysis of unknown sample) ∑¥ Õ∫‚¥¬°“√«‘‡§√“–Àåµ—«Õ¬à“߇≈◊Õ¥∑’Ë ‡µ‘¡ “√¡“µ√∞“π 1,2-‰¥§≈Õ‚√Õ’‡∏π∑’˧«“¡‡¢â¡ ¢âπ 10 mg/L µ—«Õ¬à“߇≈◊Õ¥∑’ˇµ‘¡ “√¡“µ√∞“𠉥§≈Õ‚√¡’‡∏π∑’˧«“¡‡¢â¡¢âπ 0.1 mg/L ·≈– µ—«Õ¬à“ßªí “«–∑’ˇµ‘¡ “√¡“µ√∞“π1,2-‰¥§≈Õ ‚√Õ’ ‡ ∏π∑’Ë § «“¡‡¢â ¡ ¢â π 80 mg/L ∑”°“√ «‘‡§√“–À嵫— Õ¬à“ß·µà≈–™π‘¥¥â«¬«‘∏∑’ æ’Ë ≤ — π“¢÷πÈ „À¡àπ’È 10 ´È”„π‡«≈“ 10 «—π‚¥¬«‘‡§√“–Àå«—π≈– 1 §√—Èß (·∑π°“√«‘‡§√“–À嵫— Õ¬à“ß∑’‰Ë ¡à∑√“∫§à“ ‡π◊ÕË ß®“° ‰¡à¡’®”Àπà“¬„π∑âÕßµ≈“¥)

º≈°“√»÷°…“ æ“√“¡‘‡µÕ√å∑’Ë∑”°“√∑¥ Õ∫

§à“¡“µ√∞“π

1,2-‰¥§≈Õ‚√Õ’‡∏π

‰¥§≈Õ‚√¡’‡∏π

‰µ√§≈Õ‚√Õ–´’µ‘°·Õ´‘¥

Linearity (>0.995) Range (§√Õ∫§≈ÿ¡§à“¡“µ√∞“π) Limit of Detection (LOD) Limit of Quantitation (LOQ) Precision Horwitz ûs Ratio

0.9996 1 -15 mg/L (10 mg/L) 0.125 0.183 0.806 (1 mg/L) 0.661 (10 mg/L) 0.544 (15 mg/L) 107 (1 mg/L) 97 (10 mg/L) 100.01 (15 mg/L) 8.48 (1 mg/L) 4.93 (10 mg/L) 3.80 (15 mg/L) 106

0.9992 0.01 - 4 mg/L (0.5 mg/L) 0.014 0.022 0.302 (0.1 mg/L) 0.185 (1 mg/L) 0.403 (2 mg/L) 106 (0.1 mg/L) 97 (1 mg/L) 101 (2 mg/L) 4.72 (0.1 mg/L) 1.95 (1 mg/L) 3.82 (2 mg/L) 104

0.9998 1 -160 mg/L (~75 mg/L) 0.139 0.165 0.625 (1 mg/L) 0.321 (80 mg/L) 0.216 (160 mg/L) 100.83 (1 mg/L) 100.18 (80 mg/L) 100.01 (160 mg/L) 6.55 (1 mg/L) 1.70 (80 mg/L) 1.21 (160 mg/L) 99.970

Accuracy (% Recovery) (90-110) Coefficient of variation (% CV) Analysis of unknown sample (% Recovery)

88

‡¡…“¬π - ¡‘∂ÿπ“¬π 2554

°√¡Õπ“¡—¬ à߇ √‘¡„Àâ§π‰∑¬ ÿ¢¿“楒

°“√æ—≤π“«‘∏’°“√µ√«®«‘‡§√“–Àå Biomarker ¢ÕߺŸâ —¡º— “√1,2-‰¥§≈Õ‚√Õ’‡∑𠉥§≈Õ‚√¡’‡∑π ·≈–‰µ√§≈Õ‚√‡Õ∏‘≈’πœ

√ÿª·≈–«‘®“√≥åº≈ ®“°°“√∑¥ Õ∫§ÿ ≥ ¡∫— µ‘ µà “ ßÊ „π °√–∫«π°“√¬◊π¬—𧫓¡∂Ÿ°µâÕߢÕß«‘∏’«‘‡§√“–Àå 1,2-‰¥§≈Õ‚√Õ’‡∏π„π‡≈◊Õ¥ ‰¥§≈Õ‚√¡’‡∏π„π ‡≈◊Õ¥·≈–‰µ√§≈Õ‚√Õ–´’µ‘°·Õ´‘¥„πªí “«– ¥â««‘∏’∑’Ëæ—≤π“¢÷Èππ—Èπ ‡¡◊ËÕæ‘®“√≥“µ“¡§ÿ≥ ¡∫—µ‘∑’Ë πà“‡™◊ËÕ∂◊Õ (Reliability Characteristic) ´÷Ëß®–¥Ÿ §ÿ≥ ¡∫—µ‘∑’Ë Natrella [22] „Àâæ‘®“√≥“§à“ LOD §à“ Precision ·≈–§à“º≈°“√«‘‡§√“–Àåµ—«Õ¬à“ß∑’ˉ¡à ∑√“∫§à“ æ∫«à“«‘∏’«‘‡§√“–Àå∑’ˉ¥âæ—≤π“¢÷Èπ¡“π’ȉ¥â æ‘ ®Ÿ πå·≈â««à“§à“µà“ßÊ ∑’‰Ë ¥âÕ¬Ÿ„à π‡°≥±å∑¬’Ë Õ¡√—∫‰¥â ‚¥¬¡’§à“ LOD ∑’Ë«‘‡§√“–À宓° sample blank ∑’Ë ‡µ‘¡ “√¡“µ√∞“π≈߉ª æ∫«à“«‘∏«’ ‡‘ §√“–Àå 1,2-‰¥ §≈Õ‚√Õ’‡∏π„π‡≈◊Õ¥¡’§à“ LOD ‡∑à“°—∫ 0.125 mg/L «‘∏’«‘‡§√“–À剥§≈Õ‚√¡’‡∏π„π‡≈◊Õ¥¡’§à“ LOD ‡∑à“°—∫ 0.014 mg/L ·≈–«‘∏’«‘‡§√“–À創√ §≈Õ‚√Õ–´’µ‘°·Õ´‘¥„πªí “«–¡’§à“ LOD ‡∑à“°—∫ 0.139 mg/L à«π Precision ¡’§à“ Horwitz ûs Ratio (Typical HORRAT) ´÷Ë߇ªìπ§à“∑’Ë· ¥ß „Àâ‡ÀÁπ∂÷ߧ«“¡·¡àπ¬”¢Õß«‘∏’°“√«‘‡§√“–Àå ®“° °“√»÷°…“æ∫«à“«‘∏’«‘‡§√“–Àå∑’Ëæ—≤π“¢÷Èπ∑—Èß 3 «‘∏’

¡’ §à “ ∑’Ë ®— ¥ Õ¬Ÿà „ π™à « ß 0.5 - 1.5 · ¥ß«à “ «‘ ∏’ «‘‡§√“–Àå∑—Èß 3 «‘∏’¥—ß°≈à“«„π·µà≈–√–¥—∫§«“¡‡¢â¡ ¢âπ (3 √–¥—∫§«“¡‡¢â¡¢âπ) ¡’§«“¡·¡à𬔥’¡“° ‡æ√“–§à“∑’ˉ¥â®—¥Õ¬Ÿà„π™à«ß Typical HORRAT πÕ°®“°π’È Linearity ‰¥â°√“ø¢Õß “√¡“µ√∞“π ‡ âπµ√ß¡’§“à correlation coefficient Ÿß°«à“ 0.995 ·≈–¡’§à“Õ¬Ÿà„π™à«ß∑’˧√Õ∫§≈ÿ¡§à“¡“µ√∞“π¢Õß “√·µà≈–™π‘¥ à«π§ÿ≥ ¡∫—µ‘√Õߧ◊Õ§à“ % Reco very æ∫«à“∑—Èß 3 «‘∏’∑’Ëæ—≤π“¢÷Èπ¡’ % Recovery Õ¬Ÿà„π™à«ß 90-110 % ·≈–§à“ Coefficient of variation (CV) ¡’§à“Õ¬Ÿà„π‡°≥±å∑’ˬա√—∫‰¥â ¥â«¬‡™àπ°—π ¥—ßπ—Èπ®÷ß √ÿª‰¥â«à“«‘∏’«‘‡§√“–Àå 1,2-‰¥ §≈Õ‚√Õ’‡∏π„π‡≈◊Õ¥ ‰¥§≈Õ‚√¡’‡∏π„π‡≈◊Õ¥ ·≈– ‰µ√§≈Õ‚√Õ–´’µ°‘ ·Õ´‘¥„πªí “«–¥â«¬‡∑§π‘§ Gas Chromatography Electron Capture Detection Headspace (GC-ECD-HS) ∑’Ë æ— ≤ π“¢÷È π “¡“√∂π”¡“„™â‰¥â„πÀâÕߪؑ∫—µ‘°“√ ‡æ◊ËÕµ√«® «—¥√–¥—∫°“√ —¡º— “√Õ‘π∑√’¬å‰Õ√–‡À¬ ‡æ◊ËÕ°“√ ‡ΩÑ“√–«—ß ªÑÕß°—π‚√§∑’ˇ°‘¥®“°°“√ª√–°Õ∫Õ“™’æ ·≈–®“° ‘Ëß·«¥≈âÕ¡

‡Õ° “√Õâ“ßÕ‘ß 1 Kusakabe T, Saito T, Takeichi S. Headspace solid-phase micro-extraction and gas chromatography with micro-electron capture detection for the measurement of p,pû-DDE in rat whole blood and hair.

Tokai J Exp Clin Med. 2003;28(3):131-8.

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3. Barkley J, Bunch J, Bursey JT, Castillo N, Cooper SD, Davis JM, et al. Gas chromatography mass spectrometry computer analysis of volatile halogenated hydrocarbons in man and his environment-A multimedia environmental study. Biomed Mass Spectrom. 1980;7:139-7.

4. Wallace LA, Pellizzari E, Hartwell T, Rosenzweig M, Erickson M, Sparacino C, et al. Personal exposure to volatile organic compounds: I. Direct measurements in breathing-zone air, drinking water, food and exhaled breath. Environ Res. 1984;35: 293-9.

5. Wallace L, Pellizzari E, Hartwell T, Zelon H, Sparacino C, Perritt R, et al. Concentrations of 20 volatile organic compounds in the air and drinking water of 350 residents of New Jersey compared with concentrations in their exhaled breath. J Occup Med. 1986;28: 603-8.

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