1 CLG- AGON1.04 Page 1 of 25 Contents A. INTRODUCTION... 2 B. EQUIPMENT... 2 C. REAGENTS AND SOLUTIONS... 4 D. STANDARD(S)... 4 E. SAMPLE PREPARATION... 6 F. ANALYTICAL PROCEDURE... 7 G. CALCULATIONS / IDENTIFICATION H. SAFETY INFORMATION AND PRECAUTIONS I. QUALITY ASSURANCE PLAN J. APPENDIX K. APPROVALS AND AUTHORITIES... 25
2 CLG- AGON1.04 Page 2 of 25 A. INTRODUCTION 1. Background Beta-agonists are also known as beta-adrenergic agonists. An adrenergic agent is a drug that has effects similar to epinephrine (adrenaline). Adrenergic drugs either stimulate a response (agonists) or inhibit a response (antagonists). The five categories of adrenergic receptors are: α 1, α 2, β 1, β 2, and β 3, and agonists vary in specificity between these receptors, and may be classified respectively. Thus, beta-agonists stimulate a response of the beta receptors. Beta-agonists are used for growth promotion in food animals, increasing lean muscle mass. In humans, clenbuterol and salbutamol are used as bronchodilators by asthma sufferers and as performance-enhancing drugs by athletes. Human side effects include increased heart rate and blood pressure, anxiety, palpitation, and skeletal muscle tremors. 2. Summary of Procedure Free residues of clenbuterol, salbutamol, cimaterol, zilpaterol, and ractopamine are extracted from retinal, liver, or muscle tissues with a mixture of acetonitrile and isopropanol. Sodium chloride, sodium sulfate, and magnesium sulfate salts are used to precipitate proteins and dehydrate the solution. This extract is evaporated, reconstituted in water, filtered, and analyzed by HPLC/MS/MS. 3. Applicability This method is suitable for the screening and/or confirmation of β-agonists in bovine retinal tissue (except for zilpaterol); bovine, porcine, ovine, and caprine liver; and bovine and porcine muscle. Screening is applicable at 3 ppb for clenbuterol, salbutamol, cimaterol, and ractopamine HCl; and 6 ppb for zilpaterol. Note: Refer to 21CFR for tolerance values set by FDA. B. EQUIPMENT Note: Equivalent equipment may be substituted. 1. Apparatus a. Petri Dishes disposable polystyrene, Optilux 100 x 20 mm, Cat. No. 1005, Becton Dickinson. b. Scalpels Fisherbrand single-use scalpel, Cat. No D, Fisher, or Razor Blades Single edge safety razor, American Safety Razor Co.
3 CLG- AGON1.04 Page 3 of 25 c. Cut Resistant Glove Cat. No. FSIS-50, FSIS Beltsville Service Center, or Ansell Polar Bear Glove, Cat. No , Fisher. d. Waring commercial blender Model 51BL31. e. Tissuemizer Polytron, Kinematica AG, Model No. PT2100. f. Top loading balance 0.01 g sensitivity, PJ3600 DeltaRange, Mettler. g. Centrifuge tubes 50 ml round bottom polyallomer with sealing caps, Cat. No , Nalgene. h. Centrifuge tubes 50 ml conical, disposable, polypropylene, with caps, Cat. No , Becton Dickinson. i. Micropipettes Adjustable, µl, Eppendorf. j. Glass rods 10 mm diameter by 25 cm length, fired and rounded at both ends. k. Vortex mixer variable speed, Cat. No. S8223-1, American Scientific Products. l. Shaker Horizontal flatbed, two speed, Cat. No , Eberbach. m. Centrifuge International Equipment Company B-22M high speed with rotor 876 for 50 ml tubes, Cat. No , VWR Scientific. n. Syringeless filter device Mini-UniPrep, 0.45 µm nylon, Cat. No. UN203NPUNYL, Whatman. o. Evaporator N-Evap 112, Organomation Associates Inc., Model No p. Filter 0.45 µm, nylon, acrodisc-13, Cat. No. 4551, Pall Gelman Sciences, Inc. q. Amber autosampler vial, 12 x 32 mm, E&K, Cat. No. E r. Analytical balance g sensitivity, AG204, Mettler. s. Volumetric flasks 1 L, 100 ml amber, 10 ml amber. t. HPLC mobile phase filtering and degassing apparatus Microfiltration Assembly, 47 mm, Millipore. u. HPLC guard column - C18 Security Guard cartridge, 2.1 x 4 mm, 2 µm particles, Cat. No. AJO-4286 and Security Guard kit, Cat. No. KJO-4282, Phenomenex. v. HPLC column - BetaMax Base, 2.1 x 100 mm, 5 µm particles, Cat. No , Thermo Hypersil. w. Food processor Robot Coupe model RSI6Y-1, Robot Coupe USA Inc. x. Sample cups evalue 4.5 oz specimen containers with caps, Cat. No. C686550, E&K Scientific.
4 CLG- AGON1.04 Page 4 of Instrumentation a. Waters Alliance 2695 HPLC equipped with an autosampler. b. Waters Micromass Quattro micro API mass spectrometer. C. REAGENTS AND SOLUTIONS Note: Equivalent reagents / solutions may be substituted. The stability time frame of the solution is dependant on the expiration date of the components used or the listed expiration date, whichever is soonest. 1. Reagents a. Methanol (MeOH) HPLC grade, Cat. No , Burdick & Jackson. b. Acetonitrile (ACN) HPLC grade, Cat. No , Burdick & Jackson. c. Isopropanol (IPA) HPLC grade, Cat. No. AH323-4, Burdick & Jackson. d. Sodium chloride (NaCl) ACS reagent grade, Cat. No. S271-1, Fisher. e. Sodium sulfate (Na 2 SO 4 ) ACS reagent grade, Cat. No , Acros Organics. f. Magnesium sulfate (MgSO 4 ) anhydrous, minimum 99.5% purity, Cat. No. M-7506, Sigma. g. Water Deionized, HPLC grade, Millipore Rx system. h. Formic acid Purity %, Cat. No , Riedel-de Haën. 2. Solutions a. Aqueous mobile phase (0.1% formic acid in water): D. STANDARD(S) Pipette 1 ml of formic acid into a 1 L volumetric flask. Fill to volume with Millipore water. Degas before use. Vacuum filter if desired through a 0.45 µm nylon filter. This solution is stable for 1 month at room temperature. Note: Equivalent standards / solutions may be substituted. The stability time frame of the solution is dependant on the expiration date of the components used or the listed expiration date, whichever ends sooner. 1. Standard Information a. Clenbuterol (CLEN) HCl approximately 95% pure, Cat. No. C-5423, Sigma Chemical Co. b. Cimaterol (CIM) Cat. No , MP Biochemicals Inc.
5 CLG- AGON1.04 Page 5 of 25 c. Salbutamol (SAL) approximately 95% pure, Cat. No. S-8260, Sigma Chemical Co. d. Ractopamine HCl (RAC) Elanco Animal Health. e. Zilpaterol (ZIL) Intervet Inc. 2. Preparation of Standard Solution(s) Note: Adjust all standard weights for purity. The concentration of Clenbuterol HCl is to be corrected for salt, since Clenbuterol is the analyte of interest as opposed to Clenbuterol HCl. The Ractopamine HCl concentration is not to be corrected for salt since Ractopamine HCl is the analyte of interest. See calculation in section G. a. CLEN, CIM, SAL and ZIL stock standards (~25 µg/ml): Weigh ~2.5 mg of CLEN, CIM, SAL and ZIL into its own 100 ml amber volumetric flask and bring to volume with methanol. Record the weight to 0.1 mg and calculate the exact concentration. These standards are stable for 2 months when stored in a refrigerator at 2-8 C. b. RAC stock standard (~1 mg/ml): Weigh ~10.0 mg into a 10 ml volumetric flask and dilute to volume with methanol. Record the weight to 0.1 mg and calculate the exact concentration. This standard is stable for 3 months when stored in a refrigerator at 2-8 C. c. RAC intermediate standard (10 µg/ml): Pipette ~100 µl (adjusted for the actual stock standard concentration) of the RAC stock standard into a 10 ml volumetric flask and bring to volume with Millipore water. This standard is stable for 1 month when stored in a refrigerator at 2-8 C. d. Mixed intermediate standard A (250 ng/ml CLEN, CIM, SAL, RAC; 500 ng/ml ZIL): Pipette ~100 µl (adjusted for the actual stock standard concentration) of the CLEN, CIM, SAL stock standards, ~200 µl of the ZIL stock standard, and ~250 µl of the 10 µg/ml RAC intermediate standard into a 10 ml amber volumetric flask and bring to volume with Millipore water. This standard is stable for 1 month when stored in a refrigerator at 2-8 C. e. Mixed intermediate standard B (50 ng/ml CLEN, CIM, SAL, RAC; 100 ng/ml ZIL): Pipette 200 µl of the mixed intermediate standard A into an HPLC vial and add 800 µl Millipore water. This standard is stable for 1 month when stored in a refrigerator at 2-8 C.
6 CLG- AGON1.04 Page 6 of 25 f. Mixed external standard (3 ng/ml CLEN, CIM, SAL, RAC; 6 ng/ml ZIL): Pipette 60 µl of mixed intermediate standard B and 940 µl of Millipore water into an HPLC vial. This standard is stable for 1 month when stored in a refrigerator at 2-8 C. E. SAMPLE PREPARATION 1. Retinal excision a. Eyeballs should be thawed at room temperature just enough so that the outside tissues can be manipulated but the aqueous and vitreous humors are still frozen. This takes about 45 to 60 minutes. The eyeball should not deform any noticeable amount when squeezed. b. Incise the eyeball by slowly cutting across the cornea horizontally and vertically with a scalpel or a razor blade, using whichever provides more control during the cutting process. The length of the incisions should include the full width and length of the cornea, and extend into the sclera to allow the eyeball to be everted. Force the semi-frozen contents (aqueous and vitreous humors and lens) out of the eyeball and retain (the eye contents may be discarded if retinal results are negative for beta agonists). Caution: To ensure minimal risk from BSE while performing the excision, wear a cut resistant glove on the hand holding the eyeball, and safety glasses or a face shield. Wash hands thoroughly after performing the excision. c. Evert the eyeball, and scrape the choroid/pre layer into a Petri dish. This layer is distinctive in bovine species due to its iridescent bluish-green metallic coloration on black background. Include black filmy tissue emanating from the optic nerve area (neural retina) if observed. Note: Whole eyeballs can be stored at -20 ºC or lower for at least one year. Extracted retinal tissue can be stored at -40 ºC or lower for one week (longer results in dry tissue). d. With a second razor blade, mince the choroid/pre tissue into fine pieces prior to weighing. 2. Liver and muscle homogenization (For alternative muscle sample homogenization, see E.2.b.) a. Blender or food processer i. Cut liver or muscle sample into smaller pieces and homogenize in a blender or food processor. ii. Transfer homogenized sample into a plastic bag and store in a freezer at -10 ºC or colder.
7 CLG- AGON1.04 Page 7 of 25 iii. Let sample partially thaw prior to analysis. b. Alternatively, dry ice grinding can be used for muscle sample homogenization i. Chop lb of muscle tissue into small pieces and homogenize with an equal amount of dry ice in a large food processor. The resulting sample homogenate will be a frozen powder. ii. iii. Transfer a portion of the homogenized sample into a loosely capped sample cup until the dry ice has sublimed. Excess sample from step b.i. may be discarded. Tighten the caps and store in a freezer at -10 ºC or colder. F. ANALYTICAL PROCEDURE 1. Preparation of Controls and Samples for Retinal Tissue. a. Weigh two ± 0.02 g blank retina tissue potions into 50 ml round bottom polyallomer centrifuge tubes. b. Prepare positive and negative controls by fortifying one with 24 μl of mixed intermediate standard B (D.2.e). 2. Sample Extraction for Retinal Tissue a. Weigh 0.4 ± 0.02 g retina into a 50 ml round bottom polyallomer centrifuge tube. Include the prepared positive and negative controls in the sample set at this time. b. Add 800 µl acetonitrile and 200 µl isopropanol. Vortex for 2 minutes while pounding tissue with a glass rod to mash retinal tissues. c. Add 0.24 g NaCl and vortex for 2 minutes. d. Add 0.80 g Na 2 SO 4 and 0.10 g MgSO 4 and vortex for 2 minutes. Note: This is a suitable stopping point. Samples may be stored overnight at 2-8 C. e. Centrifuge the samples for about five minutes at approximately 2000 RCF. f. Filter extract by either of: i. Whatman mini-uniprep filter vial. (a) (b) (c) Pipette 0.5 ml of extract into Whatman mini-uniprep filter vial. Evaporate to dryness with air or nitrogen. Add 0.5 ml Millipore water to vial and filter reconstituted extract by pushing plunger-shaped cap equipped with a 0.45 µm nylon filter into vial.
8 CLG- AGON1.04 Page 8 of 25 ii. 3 ml syringe and 0.45 µm nylon filter. (a) (b) Pipette 0.8 ml of extract into a 12x75mm glass tube. Evaporate to dryness with air or nitrogen. (c) Add 0.8 ml of Milli-Q water to the glass tube, vortex for 30 seconds and filter using a 3 ml syringe and 0.45 µm nylon filter into a vial. 3. Preparation of Controls and Samples for Liver and Muscle Tissue. a. Weigh two 5.0 ± 0.1 g blank homogenized liver or muscle tissue portions into 50 ml disposable centrifuge tubes. b. Prepare positive and negative controls by fortifying one with 60 µl of the mixed intermediate standard A (D.2.d.). 4. Sample Extraction for Liver and Muscle Tissue a. Weigh 5.0 ± 0.1 g homogenized liver or muscle into a 50 ml disposable centrifuge tube. Include the prepared positive and negative controls in the sample set at this time. b. Add 4 ml acetonitrile and 1 ml isopropanol. For liver and dry ice ground muscle (E.2.b.), cap and shake or vortex for 2 minutes. For muscle homogenized without dry ice (E.2.a), tissuemize for 30 seconds. c. Add 1.2 g NaCl and shake or vortex for 2 minutes. d. Add 4 g Na 2 SO 4 and 0.5 g MgSO 4 and shake or vortex for 2 minutes. Note: This is a suitable stopping point. Samples may be stored overnight at 2-8 C. e. Centrifuge the samples for about five minutes at approximately 2000 RCF. f. Filter extract by either of: i. Whatman mini-uniprep filter vial. ii. (a) (b) (c) Pipette 0.5 ml of extract into Whatman mini-uniprep filter vial. Evaporate to dryness with air or nitrogen. Add 0.5 ml Millipore water to vial and filter reconstituted extract by pushing plunger-shaped cap equipped with a 0.45 µm nylon filter into vial. 3 ml syringe and 0.45 µm nylon filter. (a) (b) Pipette 0.8 ml of extract into a 12x75 mm glass tube. Evaporate to dryness with air or nitrogen.
9 CLG- AGON1.04 Page 9 of Instrumental Settings (c) Add 0.8 ml of Milli-Q water to the glass tube, vortex for 30 seconds and filter using a 3 ml syringe and 0.45 µm nylon filter into a vial. Note: The instrument parameters may be optimized to ensure system suitability. a. HPLC Conditions: Aqueous Mobile Phase 0.1% formic acid in water Organic Mobile Phase Acetonitrile Flow Rate 0.3 ml/min Column Temperature 25 C Injection Volume 50 µl Run Time 12 minutes b. HPLC Mobile Phase Gradient Table: Time % Aqueous % Organic 0:00 95% 5% 1:30 95% 5% 7:30 35% 65% 7:36 95% 5% 12:00 95% 5% c. Interface Conditions: Ion Mode ES+ Source Temperature 125 C Desolvation Temperature 400 C Cone Gas Flow 25 L/hr Desolvation Gas Flow 900 L/hr
10 CLG- AGON1.04 Page 10 of 25 d. MRM Parameters: Precursor Ion (m/z) Salbutamol 240 Cimaterol 220 Product Ion (m/z)* Collision Energy (ev) Clenbuterol Ractopamine Zilpaterol * Most abundant product ion is in bold e. MS Parameters: Segment # 1 2 Starting Retention Time (min) Dwell Time (sec) Capillary (kv) 1 1 Multiplier (V) Analytes SAL, CIM, & ZIL CLEN & RAC
11 CLG- AGON1.04 Page 11 of Injection sequence/sample Set a. External Standard b. Positive Control c. Water Blank d. Negative Control e. Intra-laboratory check sample (if needed) f. Samples, up to a maximum of 48 The external standard, positive control and negative control must be re-injected after a maximum of 24 samples. The positive control shall also be re-injected at the end of the injection sequence. G. CALCULATIONS / IDENTIFICATION 1. Calculations a. Correction of Clenbuterol Concentration for Salt and Purity Mass Clenbuterol = Mass Clenbuterol HCl x Molecular Mass Clenbuterol x Purity Where Mass Clenbuterol HCl = ~2.5 mg Molecular Mass Clenbuterol = g/mol Molecular Mass Clenbuterol HCl Molecular Mass Clenbuterol HCl = g/mol Purity = 0.95 (for the product listed) b. Estimated Amount Found This is a quantitative estimate calculated for comparison to the screen cutoff level. It is based on a one point calibration with the positive control injected most recently before the sample as the reference. The MS instruments can be programmed to automatically do this calculation. D = E * B sample / B pos. ctrl. Where D = Estimated Amount Found in the Sample (ppb) E = Positive Control Fortification Level (ppb) B sample = Quant Ion Peak Area in the Sample (counts)
12 CLG- AGON1.04 Page 12 of 25 B pos. ctrl. = Quant Ion Peak Area in the Positive Control injected most recently before the sample (counts) c. Screen Cutoff Level This level is used to determine if a sample is screen positive or negative. It includes two safety factors so that potential violations are not missed. F = 0.5 * G * H Where F = Screen Cutoff Level (ppb) G = Tolerance (ppb) The tolerance or action level will need to be inserted for the analyte in the product of interest. For zero and no tolerance situations, samples are screened at the positive control fortification level, which makes G = 2 * E, and F = E * H. Screening at half tolerance is the first safety factor. H = Minimum / Maximum Recovery (unitless) Take values from Table 1 below. Note: Values may be updated as more data becomes available. This is the second safety factor to ensure that violations are not missed due to variation in recovery. These values are based on the positive control having the maximum recovery and the sample having the minimum recovery. Table 1 Minimum / Maximum Recovery Values Min / Max Cmpd # Name recovery 1 Clenbuterol Cimaterol Salbutamol Ractopamine HCl Zilpaterol 0.50
13 CLG- AGON1.04 Page 13 of Screening Criteria a. The retention time of each analyte must match that of the positive control or the external standard injected most recently before the sample within 5%. b. All product ions for a given analyte must be present. The required ions are listed in F.5.d. c. Each ion must have a signal-to-noise ratio 3. d. The sample is screen positive if the estimated amount found equals or exceeds the screen cutoff level (D F). e. A water blank injected immediately after the initial positive control injection must be negative for all analytes according to criteria G.2.a.-c. above. f. Referring to the negative and positive controls injected most recently before the relevant sample, the negative control must be negative for all analytes according to criteria a.-c. above, with the quant ion peak area being 5% of the positive control injection. g. The positive controls injected closest in time before and after the relevant sample must both be positive for all analytes according to criteria b.-c. above and match the retention time of the external standard injected most recently before the relevant sample within 5%. 3. Confirmation Criteria a. The retention time must match that of the positive control or the external standard injected most recently before the relevant sample within 5%. b. Product ion abundance ratios must match that of the positive control or the external standard injected most recently before the relevant sample within a 20% relative difference. The following are representative ion ratios calculated relative to the most abundant product ion: Ratio #1 Ratio #2 SAL 148 / / 222 CIM 160 / / 143 CLEN 203 / / 259 RAC 164 / / 107 ZIL 244 / / 157 c. Each ion must have a signal-to-noise ratio 3. d. A water blank injected immediately after the initial positive control injection must be negative according to screening criteria G.2.a.-c. above.
14 CLG- AGON1.04 Page 14 of 25 e. Referring to the negative and positive controls injected most recently before the relevant sample, the negative control must be negative according to criteria a.-b. above, with the quant ion peak area being 5% of the positive control. f. The positive controls injected closest in time before and after the relevant sample must both be positive according to screening criteria G.2.b.-c. above and match the retention time of the external standard injected most recently before the relevant sample set within 5%. Note: Confirmation criteria for the negative control and positive control are required only for analytes that are to be confirmed in the sample set. H. SAFETY INFORMATION AND PRECAUTIONS 1. Required Protective Equipment safety glasses and/or face shield, disposable gloves, cut resistant gloves, lab coat. 2. Hazards Procedure Step Hazard Recommended Safe Procedures Preparation of retinal tissue using scalpel/razor blade May cause accidental Injury to hands Use cut resistant gloves and other protective equipment Methanol, acetonitrile, and isopropanol Flammable and poisonous Use reagents in an efficient fume hood away from all electrical devices and open flames. Wear gloves and protective eyewear. Formic acid Acid burns Wear protective equipment and avoid contact with skin. 3. Disposal Procedures Follow local, state and federal guidelines for disposal.
15 CLG- AGON1.04 Page 15 of 25 I. QUALITY ASSURANCE PLAN 1. Performance Standard a. Positive control is positive for all analytes using the criteria in Section G. b. Negative control is negative for all analytes using the criteria in Section G. 2. Critical Control Points and Specifications Record Sample weight of retinal tissue Sample weight of liver or muscle tissue Acceptable Control 0.4 ± 0.02 g 5.0 ± 0.1 g 3. Intralaboratory Check Samples a. System, minimum contents. i. Frequency: One per week per analyst when samples analyzed. ii. Records are to be maintained. b. Acceptability criteria. Refer to I. 1. If unacceptable values are obtained, then: i. Investigate following established procedures. ii. Take corrective action as warranted. 4. Condition upon receipt a. Liver, muscle, and retinal tissue Cold, not spoiled or rancid J. APPENDIX 1. Proposed fragmentation patterns Clenbuterol Ion (m/z) Fragment 277 M [M -H 2 O] [M -H 2 O-(CH 3 ) 2 C=CH 2 ] [M -H 2 O-(CH 3 ) 2 C=CH 2 -Cl ] +
CONFIRMATION OF ZOLPIDEM BY LIQUID CHROMATOGRAPHY MASS SPECTROMETRY 9.1 POLICY This test method may be used to confirm the presence of zolpidem (ZOL), with diazepam-d 5 (DZP-d 5 ) internal standard, in
CLG-AVR1.03 Page 1 of 18 Contents A. INTRDUCTIN... 2 B. EQUIPMENT... 2 C. REAGENTS AND SLUTINS... 3 D. STANDARDS... 4 E. SAMPLE PREPARATIN... 5 F. ANALYTICAL PRCEDURE... 5 G. CALCULATINS... 11 H. SAFETY
METHOD OF ANALYSIS N methyl 2-pyrrolidone Center for Veterinary Medicine U.S. Food and Drug Administration 7500 Standish Place, Rockville, Maryland 20855 September 26, 2011 This method of detection for
CLG-CEF2.00 Page 1 of 13 Contents A. INTRODUCTION... 2 B. EQUIPMENT... 2 C. REAGENTS AND SOLUTIONS... 3 D. STANDARD(S)... 4 E. SAMPLE PREPARATION... 6 F. ANALYTICAL PROCEDURE... 6 G. CALCULATIONS / IDENTIFICATION...
Food Safety and Inspection Service, ffice of Public Health Science CLG-MAL1.02 Page 1 of 20 Contents A. INTRDUCTIN... 2 B. EQUIPMENT... 2 C. REAGENTS AND SLUTINS... 3 D. STANDARDS... 4 E. SAMPLE PREPARATIN
SUCRALOSE Prepared at the 41st JECFA (1993), published in FNP 52 Add 2 (1993). Metals and arsenic specifications revised at the 63rd JECFA (2004). An ADI of 0-15 mg/kg bw was established at the 37th JECFA
And in the red corner, introducing the challenger NASDAQ April 4 2011 243.47 $ 1.38 $ 0.58 % 0.07 $ 0.02 $ 39.4 % Technology DVD Digital Streamline Instant Play Internet Access Unlimited Selection No Late
Extraction, Derivatization and Ultra-Sensitive GC/Triple Quadrupole Analysis of Estrone and Estradiol in uman Serum Technical verview Clinical Research Authors Anthony Macherone, Ph.D. Agilent Technologies,
The Use of Micro Flow LC Coupled to MS/MS in Veterinary Drug Residue Analysis Stephen Lock AB SCIEX Warrington (UK) Overview A rapid, robust, sensitive and specific LC-MS/MS method has been developed for
Method 552.1 Revision 1.0 Determination of Haloacetic Acids and Dalapon in Drinking Water by SPE and GC/ECD* UCT Products: EUQAX156 (quaternary amine with Cl - counter ion, 6 ml cartridge)** CLTTP050 (CLEAN-THRU
1 Experiment 3: Extraction: Separation of an Acidic, a Basic and a Neutral Substance Read pp 142-155, 161-162, Chapter 10 and pp 163-173, Chapter 11, in LTOC. View the videos: 4.2 Extraction (Macroscale);
Mass Standards Kit for Calibration of AB SCIEX TOF/TOF Instruments Protocol 1 Product Description The Mass Standards Kit includes reagents needed to test instrument function, optimize instrument parameters,
TECHNICAL WHO/SIT/21.R3 TECHNICAL CHLORPYRIFOS Full specification WHO/SIT/21.R3 Revised 10 December 1999 1. Specification 1.1 Description The material shall consist of chlorpyrifos together with related
QSM Approval: Coating and Extraction of Honeycomb Denuders 1 INTRODUCTION 1.1 The following procedures are used for the coating of honeycomb denuders with citric acid and sodium carbonate solutions, and
LC-MS/MS Method for the Determination of Docetaxel in Human Serum for Clinical Research J. Jones, J. Denbigh, Thermo Fisher Scientific, Runcorn, Cheshire, UK Application Note 20581 Key Words SPE, SOLA,
DNA SPOOLING 1 ISOLATION OF DNA FROM ONION INTRODUCTION This laboratory protocol will demonstrate several basic steps required for isolation of chromosomal DNA from cells. To extract the chromosomal DNA,
Summary: Determination of Anabolic Steroids in Horse Urine by SPE and LC-MS/MS UCT Part Numbers: CUNAX226 - Clean-Up C8+NAX, 2mg/6mL BETA-GLUC- ml Beta-Glucuronidase Enzyme, liquid form SLAQ1ID21-3UM -
SODIUM CHLORITE New specifications prepared at the 68 th JECFA (2007) and published in FAO JECFA Monographs 4 (2007). An ADI of 0.03 mg/kg bw for chlorite was established at the 68 th JECFA (2007). SYNONYMS
Application Note AN844 Extraction of, and from Human Plasma Using EVOLUTE EXPRESS WCX Page 1 Extraction of, and from Human Plasma Using EVOLUTE EXPRESS WCX Prior to LC-MS/MS Analysis Introduction Catecholamines
Page 1 of 5 Overview Purpose & Scope This procedure describes a method to extract high quality genomic DNA from processed plant tissues (e.g., leaf, grain, or seed). Monsanto has optimized and performed
ANALYTICAL REPORT DETERMINATION OF CHOLECALCIFEROL (VITAMIN D 3 ) IN XXXXXXXXX 1. Samples: The samples were received on xxxxxxxx, 2009, for Vitamin D3 (cholecalciferol) determination. Sample ID XXXX XXXX
Technical Procedure for the Solid Phase Extraction of Acidic, Neutral and Basic Drugs for GC-MS Analysis 1.0 Purpose - This procedure specifies the required elements for the solid phase extraction of acidic,
SPE, LC-MS/MS Method for the Determination of Ethinyl Estradiol from uman Plasma Krishna Rao Dara, Dr. Tushar N. Mehta, Asia Pacific Center of Excellence, Thermo Fisher Scientific, Ahmedabad, India Application
CORESTA RECOMMENDED METHOD N 72 DETERMINATION OF TOBACCO-SPECIFIC NITROSAMINES IN SMOKELESS TOBACCO PRODUCTS BY LC-MS/MS (February 216). INTRODUCTION The CORESTA Smokeless Tobacco Sub-Group studied various
C146-E175A Technical Report Automatic Identification and Semi-quantitative Analysis of Psychotropic Drugs in Serum Using GC/MS Forensic Toxicological Database Hitoshi Tsuchihashi 1 Abstract: A sample consisting
Extraction of Cannabinoids in Marijuana and Edibles by QuEChERS UCT Part Numbers: ECQUEU750CT-MP - QuEChERS, Mylar packs containing 4 g magnesium sulfate, 1 g sodium chloride, 0.5 g sodium citrate dibasic
Issue Implementation Page 1 of 7 GC/MS Analysis of Vanillin and Ethyl Vanillin in Distilled Spirits Scope and Application This method will determine the level of vanillin and ethyl vanillin in distilled
Removal of Beta-Glucuronidase Enzyme from Urine Post-Hydrolysis to Improve Assay Performance and Column Lifetime Shahana Huq, Seyed Sadjadi, and Sky Countryman Phenomenex, Inc., 411 Madrid Ave., Torrance,
Revision No.: ZJ0002 Issue Date: Aug 7 th, 2008 HBV Quantitative Real Time PCR Kit Cat. No.: HD-0002-01 For Use with LightCycler 1.0/LightCycler2.0/LightCycler480 (Roche) Real Time PCR Systems (Pls ignore
HARVESTING AND CRYOPRESERVATION OF HUMAN EMBRYONIC STEM CELLS (hescs) OBJECTIVE: can be cryopreserved in a liquid nitrogen (LN 2 ) freezer for long-term storage. This Standard Operating Procedure (SOP)
EXPERIMENT 12: Empirical Formula of a Compound INTRODUCTION Chemical formulas indicate the composition of compounds. A formula that gives only the simplest ratio of the relative number of atoms in a compound
1 APPLICATIONS MANUAL CONTENTS Ifosfamide in blood serum... 2 Ingredients in blood serum... 3 Organophosphorus pesticides in tea leaf... 5 Sudan i ii iii iv in chilli sauce... 6 Pah in water... 7 Phenols
Analysis of the Vitamin B Complex in Infant Formula Samples by LC-MS/MS Stephen Lock 1 and Matthew Noestheden 2 1 AB SCIEX Warrington, Cheshire (UK), 2 AB SCIEX Concord, Ontario (Canada) Overview A rapid,
Automated Solid Phase Extraction (SPE) of EPA Method 1694 for Pharmaceuticals and Personal Care Products in Large Volume Water Samples Keywords Application Note ENV0212 This collaboration study was performed
Chemistry 321, Experiment 8: Quantitation of caffeine from a beverage using gas chromatography INTRODUCTION The analysis of soft drinks for caffeine was able to be performed using UV-Vis. The complex sample
Analytical Test Report Customer: Address (City, State): Purchase Order: Report Number: Project Number: Date Received: Date of Report: Test Location: Boulder, CO. Assay: Part Number: Amino Acids by HPLC
ISLATIN F CAFFEINE FRM TEA Introduction In this experiment, caffeine is isolated from tealeaves. The chief problem with the isolation is that caffeine does not exist alone in the tealeaves, but other natural
Micromass LCT User s Guide 1) Log on to MassLynx with your username & password. 2) After you have logged in, the MassLynx software will automatically run. 3) After MassLynx has come up, open your project
CHAPTER 5 SIMULTANEOUS DETERMINATION OF TELMISARTAN AND HYDROCHLOROTHIAZIDE IN TABLET DOSAGE FORM USING REVERSE PHASE HIGH PERFORMANCE LIQUID CHROMATOGRAPHY CHAPTER 5 Simultaneous determination of Telmisartan
Southern Blot Analysis (from Baker lab, university of Florida) DNA Prep Prepare DNA via your favorite method. You may find a protocol under Mini Yeast Genomic Prep. Restriction Digest 1.Digest DNA with
Document: AND Sol Env 08 2013 Environmental Water Testing: Surface Water, Groundwater, Hard Water, Wastewater, & Seawater Matrix specific sample preparation and testing methods for environmental waters
Supplementary Materials and Methods (Metabolomics analysis) Metabolite extraction and mass spectrometry analysis. 12 Vldlr-/- knock out and 12 wild type (WT) retinas were separately frozen at -80 ºC in
HS 1003 Heavy Metals Test 1. Purpose This test method is used to analyse the heavy metal content in an aliquot portion of stabilised hot acetic acid extract by Atomic Absorption Spectroscopy (AAS). Note:
Test Procedure for DETERMINING SULFATE CONTENT IN SOILS COLORIMETRIC TxDOT Designation: Tex-145-E Effective Date: February 2005 1. SCOPE 1.1 This method determines the soluble sulfate content of soil by
EURL-FV Multiresidue Method using Mini-Luke followed by GC-QqQ-MS/MS for Fruits and Vegetables CONTENTS 1. Aim and Scope 2 2. Short Description 2 3. Apparatus and Consumables 2 4. Chemicals 3 5. Procedure
Isolation, concentration and determination of post blast residua by liquid chromatography with DAD detector capillary electrophoresis with DAD detector Firearm fumes (gunshot residua, GSR) - various metallic
Determination of Formic Acid, Acetic Acid and Propionic Acid Contents in FAME - Blended Diesel Fuels (Draft) Water Extraction Ion Chromatography Method (WARNING The use of this standard may involve hazardous
ACETA.02-1 ACETALDEHYDE and ISOVALERALDEHYDE (Gas Chromatography) PRINCIPLE Isovaleraldehyde (IVA) and acetaldehyde are released from the syrup with the aid of dilute phosphoric acid and heat. The liberated
To make Glycerol Stocks of Plasmids ** To be done in the hood and use RNase/DNase free tips** 1. In a 10 ml sterile tube add 3 ml autoclaved LB broth and 1.5 ul antibiotic (@ 100 ug/ul) or 3 ul antibiotic
Synthesis, Isolation, and Purification of an Ester AP Chemistry Laboratory Introduction An ester is a chemical compound that is formed when an organic acid reacts with an alcohol. Esters frequently have
Mouse Insulin ELISA For the quantitative determination of insulin in mouse serum and plasma Please read carefully due to Critical Changes, e.g., Calculation of Results. For Research Use Only. Not For Use
TANNIC ACID Prepared at the 71 st JECFA (2009) and published in FAO JECFA Monographs 7 (2009), superseding specifications prepared at the 39 th JECFA (1992), published in the combined Compendium of Food
Page 1 of 7 Agencourt RNAdvance Cell v2 Total RNA Isolation from Cultured Cells Please refer to http://www.agencourt.com/technical for updated protocols and refer to MSDS instructions http://www.beckmancoulter.com/customersupport/msds/msds.asp
SODIUM CARBOXYMETHYL CELLULOSE Prepared at the 28th JECFA (1984), published in FNP 31/2 (1984) and in FNP 52 (1992). Metals and arsenic specifications revised at the 55 th JECFA (2000). An ADI not specified
A 21 a - Whey Protein Nitrogen Index GEA Niro Method No. A 21 a Revised: November 2009 1. Principle The undenaturated Whey Protein Nitrogen Index (WPN) is a measure of the heat treatment applied to the
Test Procedure for ANALYSIS OF WATER FOR CHLORIDE AND SULFATE IONS TxDOT Designation: Tex-619-J Effective Date: August 2005 1. SCOPE 1.1 Use this method to analyze water for chloride and sulfate ions to
Cantaurus, Vol. 7, 5-8, May 1999 McPherson College Division of Science and Technology Develop a Quantitative Analytical Method for low (» 1 ppm) levels of Sulfate Janet Bowen ABSTRACT Sulfate is used in
SYNONYMS INS No. 464 HYDROXYPROPYLMETHYL CELLULOSE Prepared at the 74 th JECFA (2011) and published in FAO JECFA Monographs 11 (2011), superseding the specifications prepared at the 63 rd JECFA (2004),
application Note Gas Chromatography/ Mass Spectrometry Author Timothy D. Ruppel PerkinElmer, Inc. Shelton, CT 06484 USA Opiates in Urine by SAMHSA GC/MS Introduction The United States Department of Health
A ovel Approach to Quantify Unbound Cisplatin, Carboplatin, and xaliplatin in Human Plasma Ultrafiltrate by Measuring Platinum-DDTC Complex Using LC/M/M Min Meng, Ryan Kuntz, Al Fontanet, and Patrick K.
Determining the Quantity of Iron in a Vitamin Tablet Computer 34 As biochemical research becomes more sophisticated, we are learning more about the role of metallic elements in the human body. For example,
Quantitative Liquid Chromatography Analysis of in Dairy Products Using Agilent's Compact LC and Rapid Resolution LC Application Note Food Safety Authors Hua Wu, Rong An, Yao Xiao, Zhixu Zhang, and Ping
CONTENTS I. INTRODUCTION II. SAMPLE PRE-TREATMENT a. Biological Samples b. Solid Samples: Soil, Whole Foods, Tissue c. Aqueous Samples: Water, Beverages d. Non-Aqueous Liquid III. SOLID PHASE EXTRACTION
htslabs.com email@example.com 800 541.4792 760 757.8080 760 757.9367 Improved Method for the Analysis of 31 Drugs of Abuse in Oral Fluid samples using the Thomson extreme FV by LC-MS/MS Nadine Koenig 2,
Physical and Chemical Properties and Changes An understanding of material things requires an understanding of the physical and chemical characteristics of matter. A few planned experiments can help you
Analysis of Pesticides in Vegetables Using the Agilent 1260 Infinity Analytical SFC System with Triple Quadrupole MS Detection Application Note Food Testing & Agriculture Authors Edgar Naegele and Thomas
Waters Corporation Waters 2690/5 USER & TROUBLESHOOTING GUIDE Contents 2690/5 Theory Setup procedures. Troubleshooting the 2690/5 User maintenance of the 2690/5 Spare Parts 2 2690/5 Theory 2690/5 Solvent
Isolation of Caffeine from Tea Introduction A number of interesting, biologically active compounds have been isolated from plants. Isolating some of these natural products, as they are called, can require
Experiment 12- Classification of Matter Experiment Matter can be classified into two groups: mixtures and pure substances. Mixtures are the most common form of matter and consist of mixtures of pure substances.
Nitrate DOC316.53.01066 Cadmium Reduction Method Method 8039 0.3 to 30.0 mg/l NO 3 N (HR) Powder Pillows or AccuVac Ampuls Scope and application: For water, wastewater and seawater. Test preparation Instrument-specific
Identification of a Substance by Physical Properties 2009 by David A. Katz. All rights reserved. Permission for academic use provided the original copyright is included Every substance has a unique set
Experiment 3 Separation by Solvent Extraction Objectives To separate a mixture consisting of a carboxylic acid and a neutral compound by using solvent extraction techniques. Introduction Frequently, organic
Method Development of LC-MS/MS Analysis of Aminoglycoside Drugs: Challenges and Solutions authors Angela (Qi) Shen, Ling Morgan, Marcele L. Barroso, and Xin Zhang; Tandem Labs Tuyen Nguyen; Sepracor Inc.
A High Throughput Automated Sample Preparation and Analysis Workflow for Comprehensive Forensic Toxicology Screening using LC/MS/MS AB SCIEX QTRAP 4500 LC/MS/MS System and Gerstel, Inc. MultiPurpose Sampler
Journal of Metals, Materials and Minerals, Vol.20 No.3 pp.145-149, 2010 Development of Headspace Gas Chromatography-Mass Spectrometry for Determination of Residual Monomer in Polymer Latex Nopparat THAWEEWATTHANANON
MOIST.02-1 MOISTURE (Karl Fischer) PRINCIPLE SCOPE The sample is dissolved in a solvent and then titrated with standardized Karl Fischer reagent. The titration end point is detected electrically, and the
HiPer Ion Exchange Chromatography Teaching Kit Product Code: HTC001 Number of experiments that can be performed: 5 Duration of Experiment: Protocol: 5-6 hours Storage Instructions: The kit is stable for
www.megazyme.com GLUCOSE OXIDASE ASSAY PROCEDURE K-GLOX 04/14 (200 Assays per Kit) or (1960 Auto-Analyser Assays per Kit) or (2000 Microplate Assays per Kit) Megazyme International Ireland 2014 INTRODUCTION: