FAST TOXICOLOGICAL SCREENING KIT BY LC-MS/MS – Code SC9000U

The kit for the toxicologicalforensic analysis is a screening method to analyze hundreds of compounds in positive and negative mode in urine. It is possible to determine the following molecules:

POSITIVE MODE: Amitrole Alypin Bamifylline 17 alpha Methyltestosterone Amantadine Bamipin 2Amino 5chlorobenzophenone Ambroxol Beclamide 2Amino 5nitrobenzophenone Ambucetamide Beclometasone dipropionate 2Hydroxyethylflurazepam Amezinium Befunolol 3,4 Methylenedioxyamphetamine Amidopyrin Bendiocarb 3,4 Methylenedioxyethylamphetamine Amidotrizoic Acid Benfluorex 3,4 Methylenedioxymethamphetamine Amiloride Benodanil 3,5 Diiodo tyrosine Aminodantrolene Benperidole 4Benzamidosalicyclic acid Aminoglutethimide Benproperine 6Mercaptourine Aminophenazone Bensultap 6OMonoacetylmorphine Aminopromazine Bentiromide 7Aminoclonazepam Aminorex Benzatropine 7Aminodesmethylflunitrazepam Amiodarone Benzocaine 7Aminoflunitrazepam Amiphenazole Benzoctamine 7Aminonitrazepam Amisulpride Benzododec inium 9Hydroxyrisperidone Amitr iptylin Benzoylecgonine cebutolol Amlodipine Benzthiazuron Aceclidine Ammoidin Berberine Aceclofenac Amorolfine Betamethasone 21 phosphate Acemetacin Amoxicillin Betaxolol Acepromazine Amphetamine Bevonium Ac eprometazine Amrinone Bezafibrate Acetiamin Antazoline Biperiden Acetylaminonitroprophoxybenzene Apalcillin Bisoprolol Aciclovir Apomorphine Bornaprine Actinoquinol Apophedrin Brodifacoum Adenine Apraclonidin Bromacil Adrenalone Aprinidine Bromazepam Ajmaline Atenolol Bromocriptine Alachlor Atorvastatin Brompheniramine Alclometasone 17,21 dipropionate Atraton Brotizolam Alimemazine Atrazine desethyl Bucetin Alizapride Atropine Bufexamac Allopurinol Atropinmethylbromid Bumetanide Almitrine Azatadine Bunazosin alpha Hydroxyalprazolam Azelasti ne Bunitrolol alpha Hydroxytriazolam Azintamid Bupivacaine Alprazolam Aztreonam Bupranolol Alprenolol Baclofen Buprenorphine Altretamine Bambuterol Buprofezin

1 Buspirone Clobenzepam Dienogest Butaperazine Clobutinol Diethazine Butetamate Clomethiazole Diethylcarbamazine Butoxycaine Clomipramine Difenoconazole Cafaminol Clonazepam Difenoxuron Caffeine Clonidine Diflucortolone Candesartan Clopenthixol Dihydrocodeine Caproylresorcinol Clopidogrel Dihydroergocristin Captopril Cloprednol Dihydroergotamine Carazolol Clozapine Dilazep Carbamazepine Cocaine Diltiazem Carbamazepine 10,11 epoxide Codeine Dimetindene Carbendazi m Coniine Dimetotiazine Carbinoxamine Corticosterone Dimetridazole Carbutamide Cortisone Dioxethedrin Carbuterol Cotinine Diphenamid Carteolol Coumatetralyl Diphenhydramine Carvedilol Croconazole Diponium Celiprolol Cromoglicic acid Diprophylline Cerivastatin Cyamemazine Dipyridamole Cetirizine Cyclicine Disopyramide Chinine Cyclophosphamide Dixyrazine Chlorazanil Cyclovalone Dobutamine Chlorbenzoxamine Cyp roheptadine Dorzolamide Chlorcyclizine Cytarabine Doxapram Chlordiazepoxide D3 Doxepin Doxepin Chloridazon D5 Diazepam Doxylamine Chlormadinone Acetate Dapiprazole Drofenine Chlormequat Deflazacort Dummy Chlorphenethiazine Demeton Smethylsulfone Ecgonineme thylester Chlorphen iramine Denaverine EDDP Chlorpromazine Desalkylflurazepam Embutramide Chlorpromazine Sulfoxide Desipramine Enalapril Chlorprothixene Desmedipham Enoximone Cilastatin Desmethylclobazam Ephedrine Cilazapril Desmethylclomipramine Eprosartan Cimetidine Dexamethasone Esculin Cinnarizine Dexamethasone 21 isonicotinate Esmolol Cinoxacin Dexfenfl uramine Estazolam Ciprofloxacin Dextromethorphan Ethenzamide Cisapride Dextropropoxyphene Etofenamate Citalopram Diaveridine Etomidate Clarithromycin Diazepam Etoposide Clemastine Dibutyl adipate Famotidine Clenbuterol Diclofenac Fedrilate Clobazam Dicycloverine Felbamate

2 Felodipine Guanoxan Lisinopril Fenarimol Haloperidol Lonazolac Fendiline Haloxyfop ethoxyethyl ester Loperamide Fenetylline Heptaminol Loratadine Fenfluramine Heroin Lorazepam Fenpipram ide Hexazinone Lormetazepam Fenpiprane Hexobendine Losartan Fenproporex Histamine Loxapine Fentanyl Histapyrrodine Lysergide Fenticonazole Homatropine Mafenide Fexofenadine Hordenine Maprotiline Flecainide Hydrocodone MBDB Flocoumafen Hydrocortisone Meb everine Floctafenine Hydrocortisone 21 acetate Meclizine Fluanisone Hydromorphone Meclofenamic acid Fluconazole Hydroxyzine Mecloxamine Flumazenil Hym ecromone Meclozine Flunarizine Imipramine Medazepam Flunitrazepam Indanazoline Mefenamic acid Fluoxetine Indapamine Mefenorex Flupentixol Indinavir Mefexamide Fluphenazine Indoprofen Mefloquine Flupirtine Indoramin Melatonin Flurazepam Iopodic acid Melitracen Flurochloridone Ipratropium Meloxicam Flu spirilen Iprazochrome Melperone Fluticasone Propionate Irbesartan Melphalan Fluvastadine Isoaminile Mepindolol Fluvoxamine Isoconazole Mepivacaine Fuberidazole Isoprenaline Meptazinol Furalaxyl Isoproturon Mequitazine Gabapentin Isothipendyl Mescaline Galantamine Isoxicam Mesoridazine Gallopamil Isoxsuprine Mesuximide Gemcitabine Kavain Metaclazepam Glibenclamide Ketamine Metamfepramone Glibornuride Ketoprofen Metamitron Gliclazide Ke torolac Metamphetamine Glimepiride Ketotifen Metazachlor Glipizide Lamotrigine Metenolone acetate Gliquidone Lercanidipine Metformin Griseofulvin Levocabastine Methabenzthiazuron Guaifenesin Levodopa Methadone Guanabenz Levomepromazine Methaphenilene Guane thidine Levopropylhexedrin Methapyrilene Guanfacine Lidocaine Methaqualon

3 Methazolamide Nebivolol Oxilofrine Methfuroxam Nefopam Oxitriptan Methocarbamol Nicardipine Oxitropium Methoprotryne Nicotinamide Oxomemazine Methotrex ate Nicotine Oxprenolol Methylephedrine Nifedipine Oxybuprocaine Methylphenidate Nifenazone Oxybutynin Methylscopolamine Niflumic acid Oxycodone Methylthiouracil Nimodipine Oxyfedrine Metipranolol Nimorazole Oxymetazoline Metixene Nisoldipine Oxymorphon Metoprolol NIsopropylsalicylamide Oxypertine Metronidazole Nitrazepam Papaverine Metsulfuron methyl Nitrendipine Paracetamol Metyrapone Nizatidine Paraoxon Mexiletine NMethylephed rine Paroxetine Mianserin Nomifensine Penfluridol Miconazole Norbuprenorphine Pentamidine Midazolam Nordiazepam Pentoxyverine Midodrine Nordiazepam N ethyl Perazine Milrinone Norephedrine Periciazine Minoxidil Norethisterone Perindopril Mirtazapine Norethisterone acetate Perphenazine Mizol astine Norfenefrine Pethidine Moclobemide Norfentanyl Phenacetin Modafinil Norfloxacin Phenazone Molsidomine Normorphine Phenazopyridine Monocrotophos Nortriptyline Phencyclidine Monolinuron Noscapine Phenelzine Moperone Nuarimol Pheniramine Morphine Obidoxime Phenprocoumon Morphine 3ßDglucuronide Ofloxacin Phenylephrine Moxaverine Olanzapine Phenyltoloxamine Moxisylyte Ondanset ron Phenytoin Moxonidine Opipramol Pholedrine Nabumetone Orciprenaline Physostigmine Nadolol Ornidazole Pilocarpine Naftidrofuryl Orphenadrine Pindolol Naftifine Oxadixyl Pioglitazone Nalidixic acid Oxamyl Pipamperone Nalorphine Oxatomide Pirbuterol Naloxone Oxazepam Pirenzepine Naltrexone Oxcarbazepine Piretanide Na ndrolone Oxeladin Piritramide Naphazoline Oxetacaine Piroxicam NDespropylpropafenone Oxiconazole Pizotifen

4 Practolol Risperidone Tetroxoprim Prajmalium Ritodrine Tetryzoline Pramipexole Rizatriptan THC Prazepam Ropinirole THC COOH Prazosin Ropivacaine THC OH Prednisolone Rosiglitazon Thebacon Prednisone Salbutamol Theobromine Prilocaine Salicylamide Theophylline Primaquine Scopolamine Thiabendazole Primidone Serotonin Thiamazole Procainamide Sertindole Thiamine Prochlorperazine Sertraline Thiopropazate Procyclidine Sildenafil Thioprop erazine Progesterone Simazine Thioridazine Promazine Sotalol Thiram Promethazin Spirapril Tiagabine Prometryn Stanozolol Tiapride Propafenone Sulfabenza mide Ticlopidine Propiconazole Sulfadiazine Tiemonium Propionylpromazine Sulfadoxine Tilidine Propipocaine Sulfaethidole Timolol Propiverine Sulfaguanidine Tinidazole Prop ranolol Sulfamethizole Tizanidine Propyphenazone Sulfamethoxazole Tocainide Prothiopendyl Sulfapyridine Tolazoline Protriptyline Sulfaquinoxaline Tolbutamide Pseudoephedrine Sulindac Toliprolol Psilocin Sulpiride Tolnaftate Pyranocoumarin Sultiame Tolpropamine Pyribenzamine Sumatriptan Tolycaine Pyridoxine Suxibuzone Topotecan Pyrimethamine Tadalafil Torasemide Pyritinol Talinolol Tramadol Pyrvinium Tamoxifen Tranexamic acid Quetiapine Telmisartan Tr azodone Quinapril Temazepam Triadimenol Quinine Tenocixam Triallate Ramipril Terazosine Triamterene Ranitidine Terbinafine Triazolam Raubasine Terbutaline Trifluperazine Reboxetine Terbutryn Trifluperidol Remoxipride Terconazole Triflupromazine Repa glinide Terfenadine Trimethobenzamide Reproterol Tertalolol Trimethoprim Reserpine Tetracaine Trimipramine Riluzole Tetrazepam Tripelennamine

5 Triprolidine Valdecoxib Xipamide Tritoqualine Valsartan Xylometazoline Tromantadine Vardenafil Yohimbine Tropisetron Venlafaxine Zolpidem Trospium Verapamil Zopiclone Tryptamine Vincamine Zuclopenthixol Tulobuterol Warfarin Urapidil Xantinol

NEGATIVE MODE: Azelaic Acid Ethyl sulfate 2Amino 5nitrobenzophenone Aztreonam Ethyl sulfate D5 4Benzamidosalicy clic acid Benserazid Furosemid 5(p Methylphenyl) phenylhydantoin Benzthiazide Hydrochlorothiazide 5Aminosalicylic acid Bezafibrate Morphin 3ßD glucuronide D3 6Mercaptourine Brallobarbital Pentobarbital Acetazolamide Carbenoxolone Phenobarbital Acetylsalicylic Acid Chl orothiazide Propallylonal Adenine Cimetidine THC COOH Adenosine Cinchonine THC OH Adrenalone Cortisone Triamcinolone Alpr ostadil Dinoprost Amobarbital Epinephrine Amoxicillin Ethyl glucuronide Atorvastatin Ethyl glucuronide D5

6 FAST TOXICOLOGICAL SCREENING KIT – Code SC9000U

TECHNICAL FEATURES

Principle of the Method:

After optimizing the retention times of the analytes by means of the injection of the Mix Standard in LC MS/MS, it will be possible to inject the diluted urine to determine intoxication by unknown molecules.

Components of the kit (200 tests): All reagents are ready to use and stable 3 years at 2–8 °C.

Reagent A – Reconstitution Solution, 1 x 20 ml

Reagent B – Hydrolysis Enzyme, 2 x 1 ml

Reagent C – Diluting Solution with con Internal Standard, 1 x 100 ml

Reagent D – Standard mix Solution, 1 x 1 ml

Reagent M1 – Mobile Phase M1, 8 x 500 ml

Reagent M2 – Mobile Phase M2, 8 x 500 ml

Minimum Instrumental equipment: ® Instrument API 3200 QTRAP Instrument API 4000 4500 QTRAP ® ® Instrument API 5500 QTRAP Operational Computer Optional equipment: Autosampler

Urine Collection Procedure: Take 10 ml of urine Stable for about 3 months at –20 °C Centrifuge the urine before analysis.

EUREKA srl – LAB DIVISION Head Quarter: VAT N° 01547310423 Via Enrico Fermi 25 E-mail:[email protected] 60033 Chiaravalle (AN) ITALY www.eurekaone.com Tel. +39 071 7450790 Fax + 39 071 7496579

This product meets all the requirements of Directive 98/79 / EC on in vitro diagnostic medical devices (IVD). The EC declaration of conformity is available on request.

Release N° 00 1 Fast To xicolog ical Screening Kit in urine June 2017

7 PREANALYTICAL PHASE

PREPARATION OF STANDARD MIX FOR SYSTEM SET UP:

1) Dilute 1 at 10 in Methanol the Reagent D – Standard mix Solution.

2) Prepare the following dilutions using the Reagent M1 – Mobile Phase M1. ® • For API 3200 QTRAP systems: add 100 ul of solution diluted at step 1 and add 900 L of Reagent M1 – Mobile Phase M1 • For API 4000 4500 QTRAP® systems: add 10 ul of solution diluted at step 1 and add 90 ul of Methanol and 900 L of Reagent M1 – Mobile Phase M1 ® • API 5500 QTRAP systems: add 2 ul of solution diluted at step 1 and add 98 ul of Methanol and 900 L of Reagent M1 – Mobile Phase M1

INJECTION : • Inject 10 30 ul

This standard should be injected at the beginning of each session as a System Suitability Test to check the instrumental and chromatographic performance.

ANALYTICAL PROCEDURE

Enzyme reconstitution

Reagent B – Enzyme of Hydrolysis it should be reconstituted with 9,0 ml of Reagent A – Reconstitution Solution. After reconstitution, the enzyme is stable 1 years at 2–8 °C.

PREPARATION OF URINE SAMPLES

STEP 1 : Sample Hydrolysis

In eppendorf dispense in sequence:

• 500 ul of SAMPLES + 100 ul of Enzyme previously reconstituted.

STEP 2 : Incubate at 60 °C for 3 hours. Cooling

STEP 3 : Take 100 µl of hydrolized and add 500 µl of Reagent C – Diluting + Internal Std

INJECTION :

• Injection 30 ul

Release N° 001 Fast Toxicological Screening Kit in urine June 2017

8

FAST TOXICOLOGICAL SCREENING KIT – Warnings

(THIS KIT HAS BEEN DEVELOP FOR SCIEX LC/MS SYSTEMS QTRAP CONFIGURATION)

INSTRUMENTAL CONDITIONS: Cromatografic Gradient

Gradient Profile Time (min) Flow Rate A (%) B (%) (µL/min) 0.00 500 90 10 10.00 1000 10 90 15.00 1000 10 90 15.50 500 90 10 18.00 500 90 10

Flow rate: 0.5 mL/min Column oven temperature: 40°C

Separation Column Drug Screen and Quant Analytical Column, 5 m 60 Å, PFP Propyl Column, 50 x 2.1 mm (P/N 9169552)

CODE ACCESSORIES DESCRIPTION PACKAGING Drug Screen and Quant Analytical Column, 5 m 60 SRE9169552 1 PK Å, PFP Propyl Column, 50 x 2.1 mm SK9001 PEEK Tubing O.D. × I.D. 1/16 in. × 0.005 in., red 1 PK PEEK Tubing O.D. × I.D. 1/16 in. × 0.010 in., blue Peek fitting Onepiece fingertight F130X S90199511 Prefilters Javelin 1 x 10 PK

9 FAST TOXICOLOGICAL SCREENING KIT (Reference Chromatograms/Spectra)

10 FAST TOXICOLOGICAL SCREENING KIT (Reference Chromatograms/Spectra)

11