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1 Application Guide CLINICAL RESEARCH Complete solutions for vitamins, steroids, hormones, pain management drugs, therapeutic drug monitoring, and more Simplified, rapid cleanup procedures Accurate, reproducible results

2 Here to Support You with Your Clinical Research Personalized Support Your lab has its very own Technical Consultant that is available to assist with method development, optimization, troubleshooting, and product recommendations. Feel free to give them a call with any questions you may have! Get to Know Your Technical Consultant Support@phenomenex.com Take Advantage of Our Lab Our customers asked us for more support, we responded with PhenoLogix sm, a full-service analytical support laboratory. Let our PhenoLogix clinical research gurus save your lab time and money. Method development and optimization in our PhenoLogix laboratory On-site method development On-site hands-on training Explore PhenoLogix 24 Hour Access to Information Technical Database Access our vast database of technical notes, applications, webinars, and more. Search by industry, document type, and separation mode to easily find what you need. Tools Utilize our web tools to find product recommendations, create customized methods, and more. For Research Use Only. Not for use in diagnostic procedures. 214 Phenomenex, Inc. All rights reserved.

3 TABLE OF CONTENTS Steroids/Hormones and Related Compounds Aldosterone...4 Catecholamines...5 Corticosteroids Estrogens Histamine...1 Metanephrine and Normetanephrine OH Progesterone Steroids Panel Testosterone...16 Therapeutic Drug Monitoring Antiepileptic Drugs...17 Antipsychotic Drugs...18 Digitoxin and Digoxin...19 Gabapentin...2 Tricyclic Antidepressants...21 Pain Panel/Pain Management Drugs Pain Panel Amphetamines...24 Benzodiazepines...25 Opiates...26 Vitamin Testing 25-OH Vitamin D2 and D OH Vitamin D and -epi-25-oh Vitamin D...28 Methylmalonic Acid (MMA)...29 Vitamin B6... Vitamin C...1 Forensic/Drugs of Abuse Nicotine and its Metabolites...2 Bath Salts... Ordering Information Sample Preparation HPLC/UHPLC Columns Looking for something else? Let us know, we can help you find a solution. By Support@phenomenex.com Phenomenex l WEB:

4 Steroids/Hormones and Related Compounds Aldosterone from Serum Sample Preparation Pretreatment: Into individually labeled test tubes, combine.5 ml serum (or calibrator or QC sample), 1 ml 25 mm Ammonium bicarbonate (ph ), and.1 ml working internal standard solution. Solid Phase Extraction (SPE): Cartridge: Strata -X-A 6 mg/ ml Part No.: 8B-S12-UBJ (see pg. 5 for additional formats) Condition: ml 1 % Methanol Equilibrate: ml 25 mm Ammonium bicarbonate Load: Pretreated sample Wash 1: ml 25 mm Ammonium bicarbonate Wash 2: ml 25 % Methanol in Water Dry: 5 min at high vacuum Elute: 2x 1 ml Ethyl acetate/isopropanol/nh 4 OH (7:2:1) Evaporate: To 5-55 C under a gentle nitrogen stream Reconstitute: 1 µl Methanol/Water (:7) containing ~1 ppm Estriol 29 1.e4 1.2e4 1.1e4 1 LC/MS/MS Conditions: Column: Kinetex 2.6 µm XB-C18 Dimension: 5 x. mm Part No.: B-4496-Y Guard: SecurityGuard ULTRA guard cartridge system Part No.: AJ-8775 Mobile Phase: A: Water B: Acetonitrile/Methanol (5:5) Flow Rate:.5 ml/min Column Temperature: 22 C Injection Volume: µl Detection: MS/MS (AB SCIEX API 5 ) Sample: 1. Aldosterone 2. Aldosterone-D7 1e App ID min Representative calibration curve for Aldosterone. The method was linear across the concentration range of 1-1 ng/dl Analyte Q1 Q2 Aldosterone Aldosterone IS (Aldosterone-D7) Analyte Area / IS Area r= ng/dl 4

5 Steroids/Hormones and Related Compounds Catecholamines by LC/UV 1874 mau LC/UV Conditions: Column: Kinetex 2.6 µm C18 Dimension: 15 x 4.6 mm Part No.: F-4462-E Guard: SecurityGuard ULTRA guard cartridge system Part No.: AJ-8768 Mobile Phase: 5 mm Ammonium formate buffer (ph )/Methanol (97:) Flow Rate: 1.2 ml/min Column Temperature: 22 C Injection Volume: 2 µl Detection: 28 nm Sample: 1. Norepinephrine 2. Epinephrine. L-DOPA 4. Dopamine 5. Serotonin 4 2 App ID min

6 Steroids/Hormones and Related Compounds Corticosteroids from Urine Sample Preparation Pretreatment: Dilute µl urine with µl DI Water with 1 µg IS (Cortisol-D4). Solid Phase Extraction (SPE): 96-Well Plate: Strata -X 6 mg/well Part No.: 8E-S1-UGB (see pg. 5 for additional formats) Condition: 1 ml Methanol Equilibrate: 1 ml Water Load: Pretreated sample Wash 1: 1 ml Water Wash 2: 1 ml 1 % Methanol in Water Elute: 2x 5 µl 2 % Formic acid in Ethyl acetate/isopropanol (85:15) Evaporate: To 5 C under a gentle nitrogen stream Reconstitute: 1 µl Ammonium acetate/ammonium acetate in Methanol (5:5) LC/MS/MS Condtions Column: Kinetex 2.6 µm Biphenyl Dimensions: 5 x. mm Part No.: B-4622-Y Guard: SecurityGuard ULTRA guard cartridge system Part No.: AJ-928 Mobile Phase: A: 1 mm Ammonium acetate in Water B: 1 mm Ammonium acetate in Methanol Flow Rate:.4 ml/min Column Temperature: 4 C Injection Volume: 1 µl Detection: MS/MS (AB SCIEX API 4 ), ESI+ 6

7 Steroids/Hormones and Related Compounds e5 6e5 5e5 4e5 e5 2e5 1e5 1. Prednisolone 2. Prednisone min 1 2 App ID Analyte Q1 Q Cortisone Cortisone Cortisol Cortisol Prednisolone Prednisolone Prednisone Prednisone Cortisol-D Cortisol-D e6. Cortisol 8e5 4. Cortisone 6e5 4e5 2e min App ID e5 4.2e5 4.e5.8e5.6e5.4e5.2e5.e5 2.8e5 2.6e5 2.4e5 2.2e5 2.e5 1.8e5 1.6e5 1.4e5 1.2e5 1.e5 8.e4 6.e4 4.e4 2.e4. 1. Prednisolone 1. Prednisolone 2. Cortisone 2. Cortisone min App ID Representative calibration curve for Cortisol. The method was linear across the concentration range of 1-2 ng/ml r = Analyte Area / IS Area ng/ml 7

8 Steroids/Hormones and Related Compounds Estrogens Estrone and Estradiol LC/MS/MS Conditions 265 Estrone Estrone-D4 Estradiol Estradiol-D App ID 265 Column: Gemini µm NX-C18 Dimension: 5 x 2. mm Part No.: B-445-B Guard: SecurityGuard guard cartridge system Part No.: AJ-867 Mobile Phase: A:.5 mm Ammonium hydroxide in Water B:.5 mm Ammonium hydroxide in Methanol Flow Rate:.55 ml/min Column Temperature: 22 C Injection Volume: 2 µl Detection: MS/MS (AB SCIEX API 4 ), ESI - Sample: 1. Estrone 2. Estrone-D4. Estradiol 4. Estradiol-D Analyte Q1 Q Estrone Estradiol Estrone-D4 (IS) Estradiol-D (IS)

9 Steroids/Hormones and Related Compounds Estrogens Estriol LC/MS/MS Conditions Estriol min 2.8 Estriol-D App ID min Column: Gemini µm C6-Phenyl Dimension: 5 x 2. mm Part No.: B-444-B Guard: SecurityGuard guard cartridge system Part No.: AJ-7914 Mobile Phase: A:.1 % 5. N Ammonium hydroxide in Water B:.1 % 5. N Ammonium hydroxide in Methanol Flow Rate:.5 ml/min Column Temperature: 22 C Injection Volume: 2 µl Detection: MS/MS (AB SCIEX API 4 ), ESI - Sample: 1. Estriol 2. Estriol-D Analyte Q1 Q Estriol Estriol-D

10 Steroids/Hormones and Related Compounds Histamine from Plasma Sample Preparation Pretreatment: Add 1.1 ml IS solution containing 4 ng/ml histamine-d4 and 1-methylhistamine-d prepared in DI water to 5 µl sample, blank, or plasma. Solid Phase Extraction (SPE): Cartridge: Strata -X-CW mg/ ml Part No.: 8B-S5-TBJ (see pg. 5 for additional formats) Condition: 1 ml 1 % Methanol Equilibrate: 1 ml DI Water Load: Pretreated sample Wash 1: 2 ml DI Water Wash 2: 2 ml Methanol Dry: 2 min at high vacuum Elute: 2x 25 µl initial mobile phase containing B/A (87:1), fortified to contain 5 % Formic acid LC/MS/MS Conditions Analyte Area / IS Area 4.e4 2.6e4.2e4 2.8e4 2.4e4 2.e4 1.6e e min 1.4e4 1.2e4 1.e min min 4.e4 2.6e4.2e4 2.8e4 2.4e4 2.e4 1.6e4 1.2e min 1.e Representative 1.1e4 calibration curve for Histamine. The method was linear across a concentration range of.1-5 ng/ml ng / ml r =.999 App ID 265 Column: Luna µm HILIC Dimension: 1 x 2. mm Part No.: D-4449-B Guard: SecurityGuard guard cartridge system Part No.: AJ-828 Mobile Phase: A: Acetonitrile/DI Water/1 mm Ammonium formate, ph.2 (5:45:5) B: Acetonitrile/1 mm Ammonium formate, ph.2 (95:5) Flow Rate:.6 ml/min Column Temperature: 22 C Injection Volume: 2 µl Detection: MS/MS (AB SCIEX API 5 ) Sample: 1. 1-Methylhistamine 2. N-Methylhistamine. Histamine Analyte Q1 Q Histamine Histamine Methylhistamine Methylhistamine N-Methylhistamine N-Methylhistamine Histamine-D Methylhistamine-D

11 Steroids/Hormones and Related Compounds Metanephrine and Normetanephrine from Plasma Sample Preparation Pretreatment: Dilute 5 µl plasma with 1 ml water. Solid Phase Extraction (SPE): Cartridge: Strata -X-CW mg/ ml Part No.: 8B-S5-TBJ (see pg. 5 for additional formats) Condition: 4 µl Methanol/Acetonitrile (5:5) Equilibrate: 4 µl DI Water Load: Pretreated sample Wash 1: 8 µl DI Water Wash 2: 2 ml Methanol/Acetonitrile (5:5) Dry: 2 min at high vacuum Elute: 2x 2 µl 5 % Formic acid in Acetonitrile/Methanol (5:5) Evaporate: To 45 C under a gentle nitrogen stream Reconstitute: 1 µl Acetonitrile/1 mm Ammonium formate, ph.2 (95:5), cap immediately to prevent evaporation LC/MS/MS Conditions Intens ity, c ps Intens ity, c ps 7.8e4 7.e4 6.e4 5.e4 4.e4.e4 2.e4 1.e4 7.8e4 7.e4 6.e4 5.e4 4.e4.e4 2.e4 1.e min min Metanephrine-D Intens ity, cps 4.e4.5e4.e4 2.5e4 2.e4 1.5e4 1.e4 5. Normetanephrine-D min Column: Luna µm HILIC Dimension: 5 x 2. mm Part No.: B-4449-B Guard: SecurityGuard guard cartridge system Part No.: AJ-828 Mobile Phase: A: Acetonitrile/1 mm Ammonium formate, ph.2 (95:5) B: Acetonitrile/Water/1 mm Ammonium formate, ph.2 (5:45:5) Flow Rate:.4 ml/min Column Temperature: 22 C Injection Volume: µl Detection: MS/MS (AB SCIEX API 5 ) Sample: 1. Metanephrine 2. Metanephrine-D. Normetanephrine 4. Normetanephrine-D Inte nsity, c ps Metanephrine min Intens ity, cp s Normetanephrine min App ID 1797 Analyte Q1 Q Metanephrine Metaneprhine-D Normetanephrine Normetanephrine-D

12 Steroids/Hormones and Related Compounds 17-OH Progesterone from Serum Sample Preparation Pretreatment: Dilute.25 ml of sample with 1 ml DI Water and.1 ml IS solution (1 ng/ml Testosterone-D and ng/ml 17-OH-Progesterone-D8 in Methanol/DI Water (5:5). Solid Phase Extraction (SPE): Cartridge: Strata -X-A mg/1 ml Part No.: 8B-S12-TAK (see pg. 5 for additional formats) Condition: 1 ml 1 % Methanol Equilibrate: 1 ml DI Water Load: Pretreated sample Wash: 6 µl 5 % Methanol Dry: 2 min at high vacuum Elute: 2x 6 µl 1 % Methanol Evaporate: To 5-55 C under a gentle nitrogen stream Reconstitute: 25 µl 2 % Acetonitrile in.1 % Formic acid 12

13 Steroids/Hormones and Related Compounds LC/MS/MS Conditions e4 2.2e4 2.e4 1.7e4 1.6e4 1.4e4 1.2e4 1.e min 4.5 min App ID 2652 Column: Gemini µm NX-C18 Dimension: 5 x 2. mm Part No.: B-445-B Guard: SecurityGuard guard cartridge system Part No.: AJ-867 Mobile Phase: A:.1 % Formic acid in DI Water B:.1 % Formic acid in Acetonitrile Flow Rate:.4 ml/min Column Temperature: 22 C Injection Volume: 25 µl Detection: MS/MS (AB SCIEX API 4 ) Sample: 1. Testosterone OH Progesterone Analyte Q1 Q 17-OH-Progesterone OH-Progesterone OH-Progesterone-D OH-Progesterone-D Representative calibration curve for 17-OH Progesterone. The method was linear across the concentration range of 1-1, pg/ml Analyte Area / IS Area r = e 4 pg / ml 1

14 Steroids/Hormones and Related Compounds Steroids Panel LC/MS/MS Conditions E E E6 1.2E6 1.E6 8.E5 8 6.E5 4.E5 2.E App ID min Column: Kinetex 2.6 µm C18 Dimension: 1 x. mm Part No.: D-4462-Y Guard: SecurityGuard ULTRA guard cartridge system Part No.: AJ-8775 Mobile Phase: A: Water B: Methanol Flow Rate:.55 ml/min Column Temperature: 22 C Injection Volume: 2 µl Detection: MS/MS (AB SCIEX API 4 ) Sample: 1. DHEAS 2. Cortisol. Corticosterone Deoxycortisol 5. Androstenedione 6. Estradiol 7. Testosterone OH Progesterone 9. DHEA 1. Progesterone OH Vitamin D 14

15 Steroids/Hormones and Related Compounds Analyte Q1 Q Q DHEAS Cortisol Corticosterone Deoxycortisol Androstenedione Estradiol (water loss) Testosterone OH Progesterone DHEA Progesterone OH Vitamin D Representative calibration curves for Testosterone, 17-OH Progesterone, Progesterone, and Estradiol. The method was linear across a concentration range of.1-2. ng/ml r = r = Testosterone OH Progesterone Analyte Area / IS Area % CV < 7.56% % accuracy > 94.% Analyte Area / IS Area % CV < 9.8% % accuracy > 94.6% ng/ml ng/ml r = r = Progesterone Estradiol Analyte Area / IS Area Analyte Area / IS Area % CV < 8.45% % accuracy > 94.4% % CV < 9.88% % accuracy > 94.2% ng/ml ng/ml 15

16 Steroids/Hormones and Related Compounds Total Testosterone from Plasma Sample Preparation Pretreatment: Dilute.25 ml of sample with 1 ml DI Water and.1 ml IS solution (2 ng/ml Testosterone-D in methanol). Solid Phase Extraction (SPE): Cartridge: Strata -X-A mg/ ml Part No.: 8B-S12-TBJ (see pg. 5 for additional formats) Condition: 1 ml 1 % Methanol Equilibrate: 1 ml DI Water Load: Pretreated sample Wash 1: 6 µl 5 % Methanol Dry: 2 min at high vacuum Elute: 2x µl 1 % Methanol Evaporate: To 5-55 C under a gentle nitrogen stream Reconstitute: 5 µl 25 % Hydroxylamine to form oxime and heat for min at 6-65 C then add 2 µl 5 % Formic acid in DI Water.6e5.4e5.2e5.e5 2.8e5 2.6e5 2.4e5 2.2e5 2.e5 1.8e5 1.6e5 1.4e5 1.2e5 1.e5 8.e4 6.e4 4.e4 2 1 LC/MS/MS Conditions Column: Gemini µm NX-C18 Dimension: 5 x 2. mm Part No.: B-445-B Guard: SecurityGuard guard cartridge system Part No.: AJ-867 Mobile Phase: A: 1 mm Ammonium formate in Water with.1 % Formic acid B: 1 mm Ammonium formate in Acetonitrile with.1 % Formic acid e min 9 9 Flow Rate:.4 ml/min Column Temperature: 5 C Injection Volume: 25 µl Detection: MS/MS (AB SCIEX API 5 ) Sample: 1. Testosterone 2. Testosterone-D. Epitestosterone App ID 2844 Representative calibration curve for Testosterone. The method was linear across the concentration range 1-1 ng/dl Analyte Area / IS Area Analyte Q1 Q Testosterone Testosterone Testosterone-D Testosterone-D e4 ng/dl Note: the quantitation is based on the testosterone-oxime derivative See Performance Characteristics of a Novel Tandem Mass Spectrometry Assay for Serum Testosterone. Kushnir, Mark M, et. al. ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT 8418, USA. Clin Chem 52: Jan. Therapeutic Drug Monitoring 16

17 Therapeutic Drug Monitoring Antiepileptic Drugs LC/MS/MS Conditions e5.8e5.6e5.4e5.2e5.e5 2.8e5 2.6e5 2.4e5 2.2e5 2.e5 1.8e5 1.6e5 1.4e5 1.2e5 1.e5 8.e4 6.e4 4.e4 2.e App ID min Column: Kinetex 2.6 µm Biphenyl Dimension: 5 x 2.1 mm Part No.: B-4462-AN Guard: SecurityGuard ULTRA guard cartridge system Part No.: AJ-929 Mobile Phase: A: Water with.1 % Formic acid B: Methanol with.1 % Formic acid Flow Rate:.5 ml/min Column Temperature: 4 C Injection Volume: 1 µl Detection: MS/MS (AB SCIEX Triple Quad 45) Sample: 1. Pregabalin 2. Gabapentin. Levetiracetam 4. Lamotrigine 5. Zonisamide 6. Lacosamide 7. MHD 17

18 Therapeutic Drug Monitoring Antipsychotic Drugs LC/MS/MS Conditions e5 6.e e5 5.e5 4.5e5 4.e5.5e5.e5 2.5e e5 1.5e5 1.e5 5.e App ID min Column: Kinetex 2.6 µm Biphenyl Dimension: 5 x 2.1 mm Part No.: B-4462-AN Guard: SecurityGuard ULTRA guard cartridge system Part No.: AJ-929 Mobile Phase: A: Water with.1 % Formic acid B: Methanol with.1 % Formic acid Flow Rate:.5 ml/min Column Temperature: 4 C Injection Volume: 1 µl Detection: MS/MS (AB SCIEX Triple Quad 45) Sample: 1. Olanzapine 2. Clozapine. 9-OH-Risperidone 4. Risperidone 5. Norquetiapine 6. Quetiapine 7. Ziprasidone 8. Aripiprazole 18

19 Therapeutic Drug Monitoring Digitoxin and Digoxin from Plasma Sample Preparation Pretreatment: Dilute 1 ml plasma with 2 ml DI water and spike with 1 µl of 4 ng/ml oleandrin IS. Solid Phase Extraction (SPE): Cartridge: Strata -X mg/ ml Part No.: 8B-S1-TBJ (see pg. 5 for additional formats) Condition: 2 ml 1 % Methanol Equilibrate: 2 ml 1 mm Ammonium acetate Load: ml Pretreated sample Wash: 1 ml 1 mm Ammonium acetate/1 mm Ammonium acetate in Methanol (5:5) Dry: 1 min at high vacuum Elute: 2x 6 µl 1 % Methanol Evaporate: To 5-55 C under a gentle nitrogen stream Reconstitute: 1 µl Methanol/Water (5:5) LC/MS/MS Conditions min App ID Column: Kinetex 2.6 µm C8 Dimension: 5 x 2.1 mm Part No.: B-4497-AN Guard: SecurityGuard ULTRA guard cartridge system Part No.: AJ-8784 Mobile Phase: A: 1 mm Ammonium acetate B: 1 mm Ammonium acetate in Methanol Flow Rate:.4 ml/min Column Temperature: C Injection Volume: 4 µl Detection: MS/MS (AB SCIEX API 4 ) Sample: 1. Digoxin 2. Oleandrin. Digitoxin Analyte Q1 Q Digoxin Oleandrin (IS) Digitoxin

20 Therapeutic Drug Monitoring Gabapentin from Plasma Sample Preparation Pretreatment: 1. Transfer 1 µl plasma to a.5 ml centrifuge tube 2. Add 1 µl IS/1 % TCA solution to the centrifuge tube, mix briefly. 14, rpm for 1 min 4. Transfer 1 µl supernatant to an autosampler vial and QS to 1 ml with water. LC/MS/MS Conditions e5 2.e5 1.8e5 1.6e5 1.4e5 1.2e5 1.e5 8.e4 6.e4 4.e4 2.e min App ID 2649 Column: Kinetex 2.6 µm C18 Dimension: 5 x 2.1 mm Part No.: B-4462-AN Guard: SecurityGuard ULTRA guard cartridge system Part No.: AJ-8782 Mobile Phase: A: 5 mm Ammonium formate B: Acetonitrile Flow Rate:.4 ml/min Column Temperature: 22 C Injection Volume: 4 µl Detection: MS/MS (AB SCIEX API ) Sample: 1. D4-Gabapentin 2. Gabapentin 2

21 Therapeutic Drug Monitoring Tricyclic Antidepressants from Plasma Sample Preparation Pretreatment: Dilute 5 µl plasma with 1 ml 1 mm Sodium acetate buffer (ph 5) Solid Phase Extraction (SPE): Cartridge: Strata -X-Drug B mg/ ml Part No.: 8B-S128-TBJ (see pg. 4 for additional formats) Condition: NOT REQUIRED Equilibrate: NOT REQUIRED Load: 5 µl Pretreated sample Wash 1:.8 ml 1 mm Sodium acetate, ph 5 Wash 2:.8mL Methanol Dry: 8-1 min at high vacuum Elute:.4 ml Methanol/Acetonitrile (5:5) plus 2% Ammonium hydroxide Evaporate: To 45 C under a gentle nitrogen stream Reconstitute: 5 µl initial mobile phase 297 LC/MS/MS Conditions 6.7e5 6.5e5 7 6.e5 5.5e5 5.e5 4.5e5 4.e5.5e5.e5 2.5e5 2.e5 1.5e5 1.e5 5.e min App ID 297 Column: Kinetex 2.6 µm C18 Dimension: 5 x 2.1 mm Part No.: B-4462-AN Guard: SecurityGuard ULTRA guard cartridge system Part No.: AJ-8782 Mobile Phase: A:.1% Formic acid in Water B:.1% Formic acid in Methanol Flow Rate:.5 ml/min Column Temperature: 22 C Injection Volume: 1 µl Detection: MS/MS (AB SCIEX API 4 ), ESI+ Sample: 1. Doxepin 2. DM-Doxepin. Imipramine-D 4. Imipramine 5. Desipramine-D 6. Desipramine 7. Nortriptyline-D 8. Nortriptyline 9. Amitriptyline 1. Protriptyline-D 11. Protriptyline 12. Clomipramine-D 1. Clomipramine 14. DM-Clomipramine Analyte Q1 Q Protriptyline Nortriptyline DM-Doxepin Desipramine Protriptyline-D Nortriptyline-D Desipramine-D Amitriptyline Doxepin Imipramine Imipramine-D DM-Clomipramine Clomipramine Clomipramine-D

22 Pain Panel/Pain Management Drugs Pain Panel Sample Preparation We have provided several sample preparation methods to choose from depending on the goals of your analysis. Solid Phase Extraction (SPE): recommended when working with low concentrations of target analyte Protein Precipitation/β-Glucuronidase Removal: rapid removal of β-glucuronidase Pretreatment for SPE and Protein Precipitation: Urine Acid Hydrolysis Enzymatic Hydrolysis 1. To 2 ml urine sample, add 5 µl concentrated 1. Dilute 5 µl urine with 1 µl buffer* and 2 µl of hydrochloric acid. 1,, units/ml β-glucuronidase in a 96-well 2. Heat at 9 C for 2 hours. collection plate.. Add 2 ml of 2 mm Sodium acetate buffer (ph 4.). 2. Vortex for 5-6 seconds. 4. Add 1 ml of 6 N Potassium hydroxide (KOH). Vortex.. Incubate in a water bath at 62 C for minutes. 5. Centrifuge for 5-6 minutes at 4. If performing SPE, transfer sample to a 96-well collection 5,-5,5 rpm (2-22 C). plate or autosampler vial, seal and centrifuge for 1 6. Verify ph of sample is between minutes at 2, rpm. Load supernatant onto Strata-X- 7. Load pretreated sample directly onto Drug B sorbent (see procedure below.) If performing rapid Strata-X-Drug B sorbent (see procedure below). removal of β-glucuronidase, load sample directly onto Impact Protein Precipitation plate (see procedure below). Serum / Plasma / Saliva 1. Dilute plasma/serum :1 with 1 % Acetic acid. 2. Vortex.. Centrifuge for 5-6 minutes at,-,5 rpm (2-22 C). 4. Load pretreated sample directly onto Strata-X- Drug B sorbent (see procedure below). For Ultra Clean and Concentrated Samples *Buffer prepared by adding 8 ml deionized water and 111 ml glacial acetic acid to a 1 L volumetric flask, adjust final volume to the line with a 5 % KOH solution and mix by inversion. Strata-X-Drug B SPE Steps Procedure designed for 6 mg bed mass Strata-X-Drug B and can be adjusted for smaller or larger bed masses. Condition: Not required Equilibrate: Not required Load: Pretreated sample (see above for pretreatment recommendations) Wash: 2 ml of 1 mm Sodium acetate buffer (ph 5.) Wash 2: 2 ml Methanol Elute: 2 ml Ethyl acetate/isopropanol/ammonium hydroxide (ph 5.) (7:2:1) Evaporate: to dryness under a stream of Nitrogen at 5 C Reconstitute: 1 ml 15 % Methanol Inject: 5 μl Reduce solvent consumption; no conditioning or equilibration required! For Rapid Removal of Proteins (Ex. β-glucuronidase) Impact Protein Precipitation (PPT) Steps 1. Load µl Acetonitrile to the wells of an Impact Plate 2. Load 1 µl hydrolyzed sample directly onto the Impact Plate. Vortex for 2 minutes at maximum possible speed 4. Apply vacuum at 2-7 inches Hg for 2- minutes and collect filtrate in a collection plate New long drip format now available! Part no. CE

23 Pain Panel/Pain Management Drugs LC/MS/MS Conditions e6 5.e6 4.8e6 4.6e6 4.4e6 4.2e6 4.e6.8e6.6e6.4e6.2e6.e6 2.8e6 2.6e6 2.4e6 2.2e6 2.e6 1.8e6 1.6e6 1.4e6 1.2e6 1.e6 8.e5 6.e5 4.e5 2.e min App ID Column: Kinetex 2.6 µm Biphenyl Dimensions: 5 x. mm Part No.: B-4622-Y Guard: SecurityGuard ULTRA Biphenyl Cartridges Part No.: AJ-928 Mobile Phase: A:.1 % Formic Acid in Water B:.1 % Formic Acid in Methanol Flow Rate:.7 ml/min Column Temperature: 4 C Injection Volume: 1 µl Detection: MS/MS (AB SCIEX API 5 ) Analyte Q1 Q Retention Time (min) Morphine Oxymorphone Hydromorphone Amphetamine Naloxone Methamphetamine Codeine MDA Oxycodone Naltrexone Hydrocodone MDMA MDEA Norfentanyl Tramadol Benzoylecgonine Meperidine Meprobamate Norbuprenorphine Fentanyl Buprenorphine Flurazepam Analyte Q1 Q Retention Time (min) Carisoprodol PCP Propoxyphene Sufentanil MAM Midazolam Normeperidine EDDP Methadone Lorazepam Clonazepam Norpropoxyphene Oxazepam Hydroxyalprazolam Nordiazepam Flunitrazepam Temazepam Alprazolam Diazepam

24 Pain Panel/Pain Management Drugs Amphetamines from Urine Sample Preparation Pretreatment: Dilute 2 ml with 1 ml 1 mm Phosphate buffer (ph 6.) spiked with IS and.5 M Sodium periodate. Mix/vortex and incubate at room temperature for 2 minutes. Solid Phase Extraction (SPE): Cartridge: Strata -X-Drug B 6 mg/6 ml Part No.: 8B-S128-UCH (see pg. 4 for additional formats) Condition: NOT REQUIRED Equilibrate: NOT REQUIRED Load: Pretreated sample Wash 1: 2 ml 1 mm Sodium acetate, ph 5 Wash 2: 2 ml Methanol Dry: 8-1 min at high vacuum Elute: 2 ml Ethyl acetate/isopropanol/ammonium hydroxide (7:2:1) Evaporate: Add µl.5 N Methanolic hydrochloride. Evaporate to 45 C under a gentle nitrogen stream. Reconstitute: 1 ml 1 % Methanol in.1 % Formic acid LC/MS/MS Conditions 7.e e5 6.e5 5.5e5 7 5.e5 4.5e5 4.e5.5e5.e e5 1 2.e5 1.5e5 1.e5 5.e App ID min Column: Kinetex 2.6 µm C18 Dimension: 5 x 2.1 mm Part No.: B-4462-AN Guard: SecurityGuard ULTRA guard cartridge system Part No.: AJ-8782 Mobile Phase: A:.1 % Formic acid in Water B:.1 % Formic acid in Methanol Flow Rate:.4 ml/min Column Temperature: 22 C Injection Volume: 2 µl Detection: MS/MS (AB SCIEX API 4 ) Sample: 1. D11-Amphetamine 2. Amphetamine. D14-Methamphetamine 4. Methamphetamine 5. D5-MDA 6. MDA 7. D5-MDMA 8. MDMA 9. D5-MDEA 1. MDEA 24

25 Pain Panel/Pain Management Drugs Benzodiazepines from Urine Sample Preparation Pretreatment: To 2 ml urine add 1 µl β-glucuronidase solution (contains 5 F units/ml Patella Vulgata in 1 mm Acetate buffer, ph 5.) and vortex. Hydrolyze for hours at 6 C. Let cool and add 1 µl of 1 mm Phosphate buffer (ph 6.). Centrifuge for 5 minutes at 5 rpm and discard pellet. Solid Phase Extraction (SPE): Cartridge: Strata -X-Drug N mg/ ml Part No.: 8B-S129-TBJ (see pg. 4 for additional formats) Condition: NOT REQUIRED Equilibrate: NOT REQUIRED Load: Pretreated sample Wash 1: 2 ml Acetonitrile/Water (2:8) Dry: 8-1 min at high vacuum Elute: 2 ml Ethyl acetate/isopropanol (85:15) Evaporate: To 45 C under a gentle nitrogen stream Reconstitute: 5 µl initial mobile phase LC/MS/MS 2219 Conditions 7.e5 6.e5 5.e5 4.e5.e5 2.e5 1.e min App ID 2219 Column: Kinetex 2.6 µm Biphenyl Dimensions: 5 x. mm Part No.: B-4622-Y Guard: SecurityGuard ULTRA Biphenyl Cartridges Part No.: AJ-928 Mobile Phase: A:.1 % Formic acid in Water B:.1 % Formic acid in Methanol Flow Rate:.6 ml/min Injection Volume: 1 µl Column Temperature: 4 C Detection: MS/MS (AB SCIEX API 5 ) Analyte Q1 Q Retention Time (min) Alprazolam 1 (quantifier) Alprazolam 2 (confirmation) Clonazepam Clonazepam Diazepam Diazepam Flunitrazepam Flunitrazepam Flurazepam Flurazepam Hydroxyalprazolam Hydroxyalprazolam Lorazepam Lorazepam Midazolam Midazolam Nordiazepam Nordiazepam Oxazepam Oxazepam Temazepam Temazepam

26 Pain Panel/Pain Management Drugs Opiates from Urine Sample Preparation Pretreatment: To 2 ml urine add 5 µl concentrated Hydrochloric acid. Heat at 9 C for 2 hours. Add 2 ml 2 mm Sodium acetate buffer (ph 4.). Add 1 ml 6 N Potassium hydroxide then vortex. Centrifuge for 5 minutes at 5 rpm. Solid Phase Extraction (SPE): Cartridge: Strata -X-Drug B 6 mg/6 ml Part No.: 8B-S128-UCH (see pg. 4 for additional formats) Condition: NOT REQUIRED Equilibrate: NOT REQUIRED Load: Pretreated sample LC/MS/MS 2218 Conditions Wash 1: 2 ml 1 mm Sodium acetate buffer, (ph 5.) Wash 2 2 ml Methanol Dry: 8-1 min at high vacuum Elute: 2 ml Ethyl acetate/isopropanol/ammonium hydroxide (7:2:1) Evaporate: To 45 C under a gentle nitrogen stream Reconstitute: 5 µl initial mobile phase 2.e6 1.e6 App ID min Column: Kinetex 2.6 µm Biphenyl Dimensions: 5 x. mm Part No.: B-4622-Y Guard: SecurityGuard ULTRA Biphenyl Cartridges Part No.: AJ-928 Mobile Phase: A:.1 % Formic acid in Water B:.1 % Formic acid in Methanol Gradient: Time (min) Flow Rate:.6 ml/min Column Temperature: 4 C Injection Volume: 1 µl Detection: MS/MS (AB SCIEX API 5 ) Analyte Q1 Q Retention Time (min) 6-MAM 1 (quantifier) MAM 2 (confirmation) Codeine Codeine EDDP EDDP Fentanyl Fentanyl Hydrocodone Hydrocodone Hydromorphone Hydromorphone Meperidine Meperidine Methadone Methadone Morphine Morphine Naloxone Naloxone Naltrexone Naltrexone Norfentanyl Norfentanyl Normeperidine Normeperidine Norpropoxyphene Norpropoxyphene Oxycodone Oxycodone Oxymorphone Oxymorphone Propoxyphene Propoxyphene Tramadol Tramadol

27 Vitamin Testing 25-OH Vitamin D2 and D from Serum Sample Preparation 1. Add 5 µl of precipitating reagent containing internal standard to a 1.5 ml centrifuge tube 2. Pipette 1 µl serum into the centrifuge tube. Vortex 2- sec 4. Inspect each tube to ensure no unmixed sample remains in the bottom of the tube. A homogenous mixture is critical. If unmixed sample remains at the bottom of the tube, dislodge by inverting and tapping, then re-vortex. 5. Centrifuge 15 min at 1, rpm 6. Transfer 1914 supernatant into a sample vial without disturbing the pellet LC/MS/MS Conditions 6.e4 5.e4 4.e4.e4 2.e4 1.e4 25-OH Vitamin D-d6 (IS) min 1.14e4 1.e OH Vitamin D min 1.6e4 1.4e4 1.2e4 1.e OH Vitamin D min App ID 1914 Column: Kinetex 2.6 µm C18 Dimension: 5 x 4.6 mm Part No.: B-4462-E Guard: SecurityGuard ULTRA guard cartridge system Part No.: AJ-8768 Mobile Phase: A:.5 % Formic acid in Water B: 5 mm Ammonium acetate with.1 % Formic acid in Water Flow Rate: 1 ml/min Column Temperature: 5 C Injection Volume: 5 µl Detection: MS/MS (AB SCIEX API 4 ) Analyte Q1 Q 25-OH Vitamin D OH Vitamin D OH Vitamin D-d

28 Vitamin Testing 25-OH Vitamin D and -epi-25-oh Vitamin D LC/MS/MS Conditions 1 2.4e6 2.2e6 2.e6 1.8e6 1.6e6 1.4e6 1.2e6 1.e6 8.e5 6.e5 4.e5 2.e5 2 App ID min Column: Kinetex 2.6 µm PFP Dimension: 1 x 2.1 mm Part No.: D-4477-AN Guard: SecurityGuard ULTRA guard cartridge system Part No.: AJ-8787 Mobile Phase: A:.1 % Formic acid in Water B:.1 % Formic acid in Methanol Flow Rate:.4 ml/min Column Temperature: 22 C Injection Volume: 2 µl Detection: MS/MS (AB SCIEX API 5 ) Sample: OH Vitamin D OH Vitamin D2. -epi-25-oh Vitamin D Analyte Q1 Q 25-OH Vitamin D OH Vitamin D/Epi-D OH Vitamin D-d OH Vitamin D (sec trans) OH Vitamin D2 (sec trans) Representative calibration curves of 25-OH Vitamin D2 and 25-OH Vitamin D. The method was linear across the concentration range of ng/ml Analyte Area / IS Area 25-OH Vitamin D ng/ml App ID 26 Analyte Area / IS Area OH Vitamin D ng/ml App ID 27 28

29 Vitamin Testing Methylmalonic Acid (MMA) from Urine or Serum Sample Preparation Pretreatment: Dilute 1 µl serum or urine with.5 ml 25 mm Ammonium formate and 5 µl IS. Solid Phase Extraction (SPE): Cartridge: Strata -X-AW mg/1 ml Part No.: 8B-S8-TAK (see pg. 5 for additional formats) Condition: 1 ml 1 % Methanol Equilibrate: 1 ml 25 mm Ammonium formate Load: Pretreated sample Wash:.5 ml Methanol/Water (5:5) Dry: 5-1 min at high vacuum Elute: 2x 6 µl 2 % NH 4 OH in Methanol Evaporate: To 45-5 C under a gentle nitrogen stream Reconstitute: 2 µl.1 % Formic acid in Water LC/MS/MS Conditions 2.6e5 2.4e e5 2.e5 1.8e5 1.6e5 1.4e5 1.2e5 1.e5 8.e4 6.e4 4.e4 2.e4 1 App ID min Column: Gemini µm C18 Dimension: 1 x. mm Part No.: D-449-Y Guard: SecurityGuard guard cartridge system Part No.: AJ-7596 Mobile Phase: A:.1 % Formic acid in Water B: 1 mm Ammonium formate in Methanol with.1 % Formic acid Flow Rate:.7 ml/min Column Temperature: 4 C Injection Volume: 1 µl Detection: MS/MS (AB SCIEX API 4 ), ESI- Sample: 1. Succinic acid 2. Methylmalonic acid (MMA) 29

30 Vitamin Testing Vitamin B6 from Plasma or Serum by HPLC-Fluorescence Sample Preparation 1. Thaw plasma samples and plasma/serum spiked calibrators or pre-manufactured calibration standards and controls at ambient temperature. Protect from light. 2. Pipette 2 µl plasma blank, calibration standards, controls and plasma samples into appropriately labeled.6 ml amber microcentrifuge tubes. a. Briefly vortex the calibrators and controls immediately prior to sampling. b. Mix the plasma samples by gentle inversion immediately prior to sampling. c. Protect the tubes from light.. Add µl of 1 mg/ml semicarbazide/glycine solution into all the tubes containing samples; cap the tubes, vortex for 15 sec. 4. Incubate in the dark at room temperature for min. 5. Uncap the tubes; add 25 µl of 2 % meta-phosphoric acid to the controls and samples. 6. Recap the tubes and vortex for sec. 7. Centrifuge for 5 min at 14, rpm at room temperature. Note: The relative centrifugal force (RCF) = 16, g. 8. Transfer 15 µl of supernatant to an amber autosampler glass Verex vial. 9. Cover the vial with a screw cap and place it in the autosampler at room temperature HPLC-Fluorescence Conditions mau min Representative calibration curve for Pyridoxal 5 -phosphate. The method was linear across the concentration range of nmol/l. App ID 2684 Column: Gemini µm NX-C18 Dimension: 1 x 4.6 mm Part No.: D-445-E Guard: SecurityGuard guard cartridge system Part No.: AJ-868 Mobile Phase: A: 2 mm Sodium phosphate and 1. ml Acetic acid in 1 L DI water, ph 6 B: Acetonitrile/Methanol (7:) Flow Rate: 1 ml/min Column Temperature: 5 C Injection Volume: µl Detection Fluorescence, Ex 6, Em 45 Sample: 1. Pyridoxal 5 -phosphate 2. 4-Pyridoxic Acid. Pyridoxal 12 1 Analyte Area / IS Area r= nmol / L

31 Vitamin Testing Vitamin C from Plasma or Serum by LC/UV Sample Preparation Rapid Protein Precipitation: 1. Add µl cold 5 % Meta-phosphoric acid (4 C) to the wells of an Impact Protein Precipitation Plate (see pg. 7 for ordering information) 2. Add 1 µl plasma/serum directly into the 5 % Meta-phosphoric acid. Mix 5 times by aspirating with same pipette tip 4. Centrifuge* the Impact plate at 5 g (with collection plate underneath) for 5 min at 4 C. Purified filtrate is collected in the collection plate. * A vacuum manifold may be used however 25 Hg vacuum pressure must be applied for up to 1 minutes or until sample is completely pulled through the Impact plate LC/UV Conditions mau 14 Vitamin C Uric Acid min Representative calibration curve for Vitamin C. The method was linear across a concentration range of Vitamin - 1 µg/ml. C in Human Plasma by Impact Plate (Kinetex 5 µm, XB-C18, 15 x 4.6 mm + Guard) App ID Column: Kinetex 5 µm XB-C18 Dimension: 15 x 4.6 mm Part No.: F-465-E Guard: SecurityGuard ULTRA guard cartridge system Part No.: AJ-8768 Mobile Phase: A:.1 % Formic acid in Water B: Acetonitrile Flow Rate:.8 ml/min Column Temperature: 22 C Injection Volume: µl Detection 245 nm Sample: 1. Vitamin C (ascorbic acid) 2. Uric acid Analyte Area / IS Area r= µg/ml 1

32 Forensic/Drugs of Abuse Nicotine and Metabolites from Urine Sample Preparation Pretreatment: Dilute.5 ml urine with.5 ml of 2 mm Ammonium acetate, ph 4, and 1 µl IS. Solid Phase Extraction (SPE): Cartridge: Strata -X-C 6 mg/ ml Part No.: 8B-S29-UBJ (see pg. 5 for additional formats) Condition: 2 ml 1 % Methanol Equilibrate: 2 ml Ammonium acetate buffer Load:.5 ml Pretreated sample Wash 1: 2 ml Ammonium acetate buffer Wash 2 2 ml % Methanol in Water Dry: 5 min at high vacuum Elute: 2x 2 ml 1.5 % Ammonium hydroxide in Methanol Evaporate: To 55 C under a gentle nitrogen stream Reconstitute: 5 µl Acetonitrile/2 mm Ammonium bicarbonate (1:9) LC/MS/MS Conditions 1.6e5 1.5e5 1.4e5 1.e5 1.2e5 1.1e5 1.e5 9.e4 8.e4 7.e4 6.e4 5.e4 4.e4.e4 2.e4 1.e min Analyte Q1 Q Nicotine Nicotine-D Cotinine Cotinine-D Nornicotine Nornicotine-D OH-cotinine OH-cotinine-D Anabasine App ID 2222 Column: Gemini µm NX-C18 Dimension: 5 x 2. mm Part No.: B-445-B Guard: SecurityGuard guard cartridge system Part No.: AJ-867 Mobile Phase: A: 2 mm Ammonium bicarbonate B: Acetonitrile Flow Rate:.5 ml/min Column Temperature: 25 C Injection Volume: 1 µl Detection MS/MS (AB SCIEX API 4 ), ESI+ Sample: 1. Nornicotine 2. -OH-Cotinine. Anabasine 4. Cotinine 5. Nicotine Representative calibration curve for Nornicotine. The method was linear across a concentration range of 1-5 ng/ml. Analyte Area / IS Area r = ng/ml 2

33 Forensic/Drugs of Abuse Bath Salts LC/MS/MS Conditions e6 4.5e e6.5e6.e6 2.5e6 2.e6 1.5e6 1.e6 5.e min App ID 279 Column: Kinetex 2.6 µm C18 Dimension: 5 x. mm Part No.: B-4462-Y Guard: SecurityGuard ULTRA guard cartridge system Part No.: AJ-8775 Mobile Phase: A: 1 mm Ammonium formate B:.1 % Formic acid in Methanol Flow Rate:.5 ml/min Column Temperature: 22 C Injection Volume: 5 µl Detection MS/MS (AB SCIEX API 4 ) Sample: 1. Methamphetamine 2. Methylone. Mephedrone 4. MDPV

34 Solid Phase Extraction (SPE) Capitalize on Cleanliness without Sacrificing Recovery With Strata-X polymeric SPE sorbents the guesswork that leads to lengthy method development is eliminated. Unique selectivities have been developed to cover a diverse spectrum of analytes, simplifying the method development process for fast and efficient sample preparation. Strata -X SPE Sorbents Find the best Strata-X sorbent for your target analytes Acidic Compounds Neutral Compounds Basic Compounds Strong Acids (pka < 2) Strata-X-AW Weak Acids (pka 2 4) Strata-X-A Neutral Compounds Strata-X Weak Bases (pka 8 1) Strata-X-C Strong Bases (pka > 1) Strata-X-CW Reversed Phase & Weak Anion-Exchange Reversed Phase & Strong Anion-Exchange Reversed Phase Reversed Phase & Strong Cation-Exchange Reversed Phase & Weak Cation-Exchange Strata-X-Drug SPE Sorbents Choose from 2 different phases that are designed and quality control tested for drugs of abuse analysis. Ordering Information Strata-X-Drug B for BASIC drugs of abuse Format Sorbent Mass Part Number Unit Price Tab-Less Tube mg 8L-S128-TAK 1 ml (1/box) Tube 1 mg 8B-S128-AAK 1 ml (1/box) mg 8B-S128-TAK 1 ml (1/box) mg 8B-S128-TBJ ml (5/box) 6 mg 8B-S128-UBJ ml (5/box) 6 mg 8B-S128-UCH 6 ml (/box) 6 mg 8B-S128-UCL 6 ml (2/box) Giga Tube 1 mg 8B-S128-EDG 12 ml (2/box) Strata-X-Drug N for NEUTRAL drugs of abuse Format Sorbent Mass Part Number Unit Price Tube mg 8B-S129-TAK 1 ml (1/box) mg 8B-S129-TBJ ml (5/box) 6 mg 8B-S129-UBJ ml (5/box) 6 mg 8B-S129-UCH 6 ml (/box) 6 mg 8B-S129-UCL 6 ml (2/bag) 1 mg 8B-S129-ECH 6 ml (/box) 96-Well Plate 1 mg 8E-S129-AGB 2 Plates/box mg 8E-S129-TGB 2 Plates/box 6 mg 8E-S129-UGB 2 Plates/box 96-Well Plate 1 mg 8E-S128-AGB 2 Plates/box mg 8E-S128-TGB 2 Plates/box 6 mg 8E-S128-UGB 2 Plates/box SPE Method Development Tool Create a Customized Method in Under 1 Minute and Receive a FREE Sample of Strata-X 1. Visit and create a customized SPE method by inputting your target analyte, sample matrix, and sample volume. 2. After using the SPE Method Development Tool, request a free sample by clicking on the Request Free Sample button. 4 Phenomenex l WEB:

35 Ordering Information Strata -X Format Sorbent Mass Part Number Unit Price Tab-Less Tube mg 8L-S1-TAK 1 ml (1/box) 6 mg 8L-S1-UBJ ml (5/box) Tube mg 8B-S1-TAK 1 ml (1/box) mg 8B-S1-TBJ ml (5/box) 6 mg 8B-S1-UBJ ml (5/box) 1 mg 8B-S1-EBJ ml (5/box) 1 mg 8B-S1-ECH 6 ml (/box) 2 mg 8B-S1-FBJ ml (5/box) 2 mg 8B-S1-FCH 6 ml (/box) 5 mg 8B-S1-HBJ ml (5/box) 5 mg 8B-S1-HCH 6 ml (/box) 96-Well Plate 1 mg 8E-S1-AGB 2 Plates/Box mg 8E-S1-TGB 2 Plates/Box 6 mg 8E-S1-UGB 2 Plates/Box On-line Extraction Cartridge Description Part No. Unit/Box Price Strata-X on-line extraction M-S-B-CB ea cartridge, 2 x 2. mm Cartridge holder, 2 mm CH-5845 ea Strata-X-C Format Sorbent Mass Part Number Unit Price Tab-Less Tube mg 8L-S29-TAK 1 ml (1/box) 6 mg 8L-S29-UBJ ml (5/box) Tube mg 8B-S29-TAK 1 ml (1/box) mg 8B-S29-TBJ ml (5/box) 6 mg 8B-S29-UBJ ml (5/box) 1 mg 8B-S29-EBJ ml (5/box) 1 mg 8B-S29-ECH 6 ml (/box) 2 mg 8B-S29-FBJ ml (5/box) 2 mg 8B-S29-FCH 6 ml (/box) 5 mg 8B-S29-HBJ ml (5/box) 5 mg 8B-S29-HCH 6 ml (/box) 96-Well Plate 1 mg 8E-S29-AGB 2 Plates/Box mg 8E-S29-TGB 2 Plates/Box 6 mg 8E-S29-UGB 2 Plates/Box On-line Extraction Cartridge Description Part Number Unit Price Strata-X-C on-line extraction M-S48-B-CB ea cartridge, 2 x 2. mm Cartridge holder, 2 mm CH-5845 ea If Strata-X SPE products do not perform as well or better than your current SPE product of similar phase, mass and size, return your products with comparative data within 45 days and keep the Strata-X SPE product for FREE! Strata-X-CW Format Sorbent Mass Part Number Unit Price Tab-Less Tube mg 8L-S5-TAK 1 ml (1/box) Tube mg 8B-S5-TAK 1 ml (1/box) mg 8B-S5-TBJ ml (5/box) 6 mg 8B-S5-UBJ ml (5/box) 1 mg 8B-S5-ECH 6 ml (/box) 2 mg 8B-S5-FBJ ml (5/box) 2 mg 8B-S5-FCH 6 ml (/box) 5 mg 8B-S5-HBJ ml (5/box) 5 mg 8B-S5-HCH 6 ml (/box) 96-Well Plate 1 mg 8E-S5-AGB 2 Plates/Box mg 8E-S5-TGB 2 Plates/Box 6 mg 8E-S5-UGB 2 Plates/Box Description Part Number Unit Price Strata-X-CW on-line extraction M-S6-B-CB ea cartridge, 2 x 2. mm Cartridge holder, 2 mm CH-5845 ea Strata-X-A Format Sorbent Mass Part Number Unit Price Tab-Less Tube mg 8L-S12-TAK 1 ml(1/box) Tube mg 8B-S12-TAK 1 ml (1/box) mg 8B-S12-TBJ ml (5/box) 6 mg 8B-S12-UBJ ml (5/box) 1 mg 8B-S12-EBJ ml (5/box) 1 mg 8B-S12-ECH 6 ml (/box) 2 mg 8B-S12-FBJ ml (5/box) 2 mg 8B-S12-FCH 6 ml (/box) 5 mg 8B-S12-HBJ ml (5/box) 5 mg 8B-S12-HCH 6 ml (/box) 96-Well Plate 1 mg 8E-S12-AGB 2 Plates/Box mg 8E-S12-TGB 2 Plates/Box 6 mg 8E-S12-UGB 2 Plates/Box Strata-X-AW Format Sorbent Mass Part Number Unit Price Tab-Less Tube mg 8L-S8-TAK 1 ml(1/box) Tube mg 8B-S8-TAK 1 ml (1/box) mg 8B-S8-TBJ ml (5/box) 6 mg 8B-S8-UBJ ml (5/box) 1 mg 8B-S8-EBJ ml (5/box) 1 mg 8B-S8-ECH 6 ml (/box) 2 mg 8B-S8-FBJ ml (5/box) 2 mg 8B-S8-FCH 6 ml (/box) 5 mg 8B-S8-HBJ ml (5/box) 5 mg 8B-S8-HCH 6 ml (/box) 96-Well Plate 1 mg 8E-S8-AGB 2 Plates/Box mg 8E-S8-TGB 2 Plates/Box 6 mg 8E-S8-UGB 2 Plates/Box Don t forget your vacuum manifold, collection plates, and sealing mats p. 6. Phenomenex l WEB: 5

36 Phospholipid Removal Phospholipid Removal Solutions Eliminate Ion Suppression with Phree Phospholipid Removal Products Consistently remove > 99% of phospholipids to increase LC/MS sensitivity Simultaneously remove interfering proteins No additional time required, the Phree method can be performed in the same amount of time as a protein precipitation procedure Skip the method development; one method for acids, bases, and neutrals How it Works: Remove Proteins Solvent Shielding Technology prevents dripping of organic solvent, allowing for protein precipitation within the wells of the Phree Phospholipid Removal Product. Eliminate Phospholipids The Phree sorbent selectively removes phospholipids from precipitated plasma samples. Proteins Phospholipids Target Analyte Phree Phospholipid Removal Products Part No. Description Unit Price 8B-S1-TAK Phree Phospholipid Removal 1 ml Tube 1/box 8E-S1-TGB Phree Phospholipid Removal 96-Well Plates 2/box Accessories Collection Plates (deep well, polypropylene) AH Well Collection Plate 5 µl/well 5/pk AH Well Collection Plate 1 ml/well 5/pk AH Well Collection Plate 2 ml/well 5/pk AH Well Collection Plate, 2 ml Square/Round-Conical 5/pk AH Well Collection Plate, 2 ml Round/Round, 8 mm 5/pk AH Well Collection Plate, 1 ml/well Round, 7 mm 5/pk Sealing Mats AH-8597 Sealing Mats, Pierceable, 96-Square Well, Silicone 5/pk AH-8598 Sealing Mats, Pre-Slit, 96-Square Well, Silicone 5/pk AH-861 Sealing Mats, Pierceable, 96-Round Well 7 mm, Silicone 5/pk AH-862 Sealing Mats, Pre-Slit, 96-Round Well 7 mm, Silicone 5/pk AH-86 Sealing Mats, Pierceable, 96-Round Well 8 mm, Silicone 5/pk AH-864 Sealing Mats, Pre-Slit, 96-Round Well 8 mm, Silicone 5/pk AH-762 Sealing Tape Pad 1/pk Vacuum Manifolds AH-62* SPE 12-Position Vacuum Manifold Set, for tubes ea AH-624* SPE 24-Position Vacuum Manifold Set, for tubes ea AH Well Plate Manifold, Universal with Vacuum Gauge ea *Manifolds include: Vacuum-tight glass chamber, vacuum gauge assembly, polypropylene lid with gasket, male and female luers and yellow end plugs, stopcock valves, collection rack assemblies, polypropylene needles, lid support legs. Waste container included with 12-positive manifold. If Phree Phospholipid Removal products do not perform as well or better than your current phospholipid removal products, send in your comparative data within 45 days and keep the Phree Phospholipid Removal product for FREE! 6 Phenomenex l WEB:

37 Protein Precipitation Impact Rapid Protein Precipitation Quickly cleanup sample by passing biological samples through the Impact filter Increase sensitivity of your analysis by eliminating proteins which contribute to baseline noise Increase reproducibility with the leak-free membrane, preventing premature sample breakthrough and incomplete protein precipitation How it Works: Proteins Target Analyte Impact Protein Precipitation Plates Part No. Description Unit Price Impact Precipitation Plates CE-7565 Impact Protein Precipitation, Square Well, Filter Plate, 2 ml CE-7566 Impact Protein Precipitation, Square Well, Filter Plate, 2 ml, Long Drip 2/box 2/box Impact Starter Kit for Protein Precipitation CE-821 Impact Protein Precipitation Plate (2 ea) Collection Plate 2 ml (2 ea) Sealing Mat, Santoprene (AH-8199) (2 ea) ea Don t forget your vacuum manifold, collection plates, and sealing mats p. 6. If Impact does not perform as well or better than your current protein precipitation plate with similar specifications, send in your comparative data within 45 days and keep the Impact product for FREE! Phenomenex l WEB: 7

38 Thrive with Kinetex Core-Shell Technology! Complete Scalable Solution from UHPLC to HPLC Core-Shell Technology UHPLC HPLC 1. µm 1.7 µm 2.6 µm 5 µm Incredible UHPLC efficiency and performance gains Six Highly Functional Stationary Phases Phase Selectivity Kinetex Biphenyl Excellent reversed phase and aromatic retention and selectivity Kinetex C18 Kinetex XB-C18 Kinetex C8 Kinetex Phenyl-Hexyl Kinetex HILIC 2 % higher efficiency than fully porous 1.7 μm columns General hydrophobic selectivity and increased retention of polar basic compounds Excellent hydrophobic selectivity and increased retention of polar acidic compounds Less hydrophobic than a C18 phase Greater retention and separation of aromatic hydrocarbons Increased polar retention under HILIC conditions Better Performance than Fully Porous Particles Achieve sub-2 μm performance on HPLC and UHPLC systems Solid Core μm or better efficiencies at 5 μm pressures for HPLC methods Porous Shell Using sol-gel processing techniques that incorporate nano structuring technology, a durable, homogeneous porous shell is grown on a solid silica core. This highly optimized process combined with industry leading column packing technology produces highly reproducible columns that generate extremely high plate counts. Fully Porous Kinetex Core-Shell Average Efficiency Gain with Kinetex* Fully Porous Kinetex Core-Shell Average Efficiency Gain with Kinetex* 5 µm VS 5 µm 9 % Higher 1.7 µm VS 1.7 µm 2 % Higher µm VS 2.6 µm 85 % Higher 1.7 µm VS 1. µm 5 % Higher * May not be representative of all applications Ordering Information SecurityGuard 5 μm Minibore Columns (mm) ULTRA Cartridges Phases 5 x x 2.1 /pk Biphenyl B-4627-AN D-4627-AN AJ-929 XB-C18 B-465-AN D-465-AN AJ-8782 C18 B-461-AN D-461-AN AJ-8782 C8 B-468-AN D-468-AN AJ-8784 Phenyl-Hexyl B-46-AN D-46-AN AJ-8788 for 2.1 mm ID SecurityGuard ULTRA Cartridges require holder, Part No.: AJ-9 SecurityGuard 5 μm MidBore Columns (mm) ULTRA Cartridges Phases 5 x. 1 x. /pk Biphenyl B-4627-Y D-4627-Y AJ-928 XB-C18 B-465-Y D-465-Y AJ-8775 C18 B-461-Y D-461-Y AJ-8775 C8 B-468-Y D-468-Y AJ-8777 Phenyl-Hexyl B-46-Y D-46-Y AJ-8781 for. mm ID 8 Phenomenex l WEB: