METHYLETHYLKETONE (M.E.K.) IN URINE BY GC/MS in head space Code GC10010
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1 METHYLETHYLKETONE (M.E.K.) IN URINE BY GC/MS in head space Code GC10010 BIOCHEMISTRY The MEK is presented as a colorless liquid odor similar to that of acetone, is volatile and potentially explosive. The methyl ethyl ketone is used in a variety of important industrial applications such as paint thinner nitro and vinyl film, paint removers, cements and adhesives, cleaning fluids, printing, wax solvent in lubricating oil, acrylic coatings, etc.. The MEK is a flammable and irritating compound, it can cause irritation to nose, throat and eyes. Although there is evidence of data that the MEK has carcinogenic or mutagenic, is suspected to be harmful to the fetus to high maternal doses. Low levels of chronic exposure have been shown to be due to decreased memory and deterioration of the central nervous system. The proposed method determines the injection of methyl ethyl ketone by GC-MS / headspace of a rate of urine sample previously treated in an appropriate solvent. The levels of MEK are determined on a urine sample at the beginning and end of shift. EUREKA srl LAB DIVISION VAT N info@eurekaone.com Head Quarter: Via Enrico Fermi Chiaravalle (AN) ITALY Tel Fax This product fulfills all the requirements of Directive 98/79/EC on in vitro diagnostic medical devices (IVD). The declaration of conformity is available upon request. Release N 002 MEK in urine by GC/MS-headspace February
2 TECHNICAL FEATURES Principle of the method : This method enables the methyl ethyl ketone urine by GC-MS / head space. MEK is first diluted and, after the addition of a stabilizer, is directly injected into GC. Recovery: 99% Sensitivity : about 0,01 mg/l by MS Dynamic Range: 0,01-10,0 mg/l by MS Normal Range: mg/l Components of the Kit : All reagents are ready to use and stable 3 years at 2 8 C. Reagent A Diluting Solution, 1 x 200 ml Reagent B Test Solution 1,6 mg/l, 1 x 20 ml Minimum Instrumental equipment required: GC/MS Operational Computer Optional Equipment: Autosampler Urine Collection Procedure: After work shift, collect 10 ml of urine in a suitable test tube. Stable about 60 days at 20 C. 2
3 PREANALYTICAL PROCEDURE Preparation of Test Solution STEP 1 : Dispense in a tube: 1800 µl of Reagent A Diluting Solution 200 µl of Reagent B Test Solution Inject 1000 µl of solution into gas chromatograph GC. Verify that the Test solution has retention time similar to fig. 1. If the Test is all right you can start with the analytical procedure; if not, check the functionality of the analytical system. ANALYTICAL PROCEDURE STEP 1 : Preparation of Calibration Standard Solution at 0,16 mg/l Dispense in a tube: 1800 µl of Urine free from MEK 200 µl of Reagent B Chemical Standard IMPORTANT: This Calibration Standard Solution is prepared from time to time To verify that the urine is effectively free from MEK, to prepare a Blank: 1800 µl of Urine 200 µl of H 2 O HPLC grade If the Blank urine presented the peak of MEK, proceed to identify the quantitative difference between the areas / heights of the peaks of MEK in urine enriched and blank. The samples are directly injected transfer 2 ml in GC vials. Close vials hermetically Mix at Vortex Place them in the Sampler Prepare work list and start the serie according to the system s specifications Release N 002 MEK in urine by GC/MS-headspace February
4 MEK - Warnings REAGENT B : TEST SOLUTIN MEK 1,60 mg/l SET OF GAS-CROMATOGRAPH: VF-5ms Column 30 m x 0,25 mm, 0,25 µm (conditioned) Injector s Temperature 200 C Temperature 60 C x 2 minutes + 5 C/min up to 100 C + 20 C/min up to 120 C (run 11 min) Helium Gas 1 ml/min Splitting ratio about 1:20 Injection Splitless with opening after 0,10 min Syringe Temperature: 60 C MixerTemperature: 80 C (6 minutes) no agitation SET OF MASSA DETECTOR (4000): Range of Massa Operative Method in Electronic Impact (EI) Set in SIM at Characteristic Masses Temperature of Massa: 200 C; Filament on: 1 minute CONDITIONING OF VF-5 MS COLUMN Follow the manufacturer prescription. Do not condition column/s if connected to the Massa Detector. CLEANING OF THE COLUMN Disconnect the detector. Keep the column at the highest temperature for the recommended time. (See the manufacturer s instructions) OPERATIONAL COMPUTER PARAMETERS IN CONFORMITY WITH THE SPECIFICATION OF OPERATIONAL COMPUTER SOFTWARE 4
5 MEK IN URINE (Reference Chromatograms/Spectrums GC-MS / Headspace) 800 Area: 1.218e+ No Search Coun kcounts 04 stand.cal.m ek 0.16ug-m l.sms 40:150 40: % Spectrum 1A BP: min, Scan: 162, 40:150, Ion: 2112 us, RIC: , BC A 75% % % % minutes m/z Fig. 1 : Calibration Standard Fig. 2 : Massa Spectrum of Calibration Standard R.T MEK 0,16 mg/l MOLECULAR ION: 43 5
6 MEK IN URINE (Reference Chromatograms/Spectrums GC-MS / Headspace) kcounts 05 cam p.urinario m ek 0.08ug-m l.sms 40:150 40: % Spectrum 1A BP: min, Scan: 161, 40:150, Ion: 2713 us, RIC: , BC A 75% Area: No Search Counts min 50% % % minutes m/z Fig. 3 : Urine Sample Fig. 4 : Massa Spectrum of Urine Sample R.T MEK 0,08 mg/l MOLECULAR ION: 43 6
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