Q1 Scan showing sample cleanliness of Oral fluid SPE from ntercept buffer using Starta-X-C cartridge

LC Conditions

Column

Kinetex 2.6 µm C18 100 Å, LC Column 50 x 3 mm, Ea

Brand

Kinetex

Part No

00B-4462-Y0

Phase Name

C18

Elution Type

Gradient

Mobile Phase

A: 0.1% formic aid in water

B: 0.1% formic acid in MeOH

Gradient Profile

Step No.

Time(min)

%A

%B

1

0

90

10

2

4

5

95

3

5.5

5

95

4

5.51

90

10

5

7

90

10

Flow Rate

500.00 mL/min

Temperature

22°C

Detection

Tandem Mass Spec (MS-MS) @ (22°C)

Detection Info

Sample Notes

SPE Product: Strata-X-CW, 30 mg/3 mL cartridge Part# 8E-S035-TBJ Sample pre-treatment Transfer 1 mL oral fluid collected on an applicator tip in its preservative buffer (Intercept). Leave it for 2 hours followed by centrifugation for 15 mins at 600g. Remove 0.5 mL of supernatant to combine with 1 mL 1% formic acid, mix/vortex 5-10 secs to load on Strata X-CW  30mg/3 mL cartridge. Condition: 1 mL Methanol Equilibrate: 1 mL DI Water Load: Combine 0.5mL of pretreated sample with 1 mL 1% formic acid, mix/vortex 5-10 sec and load Wash 1: 1 mL 1% Formic acid/DI Water Wash 2: 1 mL DI Water Dry Down: 5-6 minutes at maximum vacuum (20" Hg or higher) Elute: 2 x 500 µL methylene chloride/Isopropanol/30% Ammonium Hydroxide (80:18:2) Dry Down: Evaporate to dryness under gentle nitrogen and 45-50°C. Reconstitute: With 167 uL initial mobile phase SPE Product: CEREX OFX 3mg Part# 633-003I NBE, 96 well NBE plate 3mg/1Ea Sample pre-treatment Transfer 1 mL oral fluid collected on an applicator tip in its preservative buffer (Intercept). Leave it for 2 hours followed by centrifugation for 15 mins at 600g. Remove 0.5 mL of supernatant to combine with 0.3 mL 0.1M HCl, mix/vortex 5-10 secs to load on SPEware cartridge Condition: 500 uL methanol Equilibrate: 500 uL water Load: Sample from pre-treatment step Wash1: 700 uL 0.1M HCl Wash2: 2x0.8 mL D.I. water Dry: 10 mins under 45050 psi Elution: 300 uL of Dichloromethane 30%ammonium hydroxide (80/18/2) Drydown: Evaporate to dryness under gentle nitrogen and 45-50°C. Reconstitute:167 µL initial mobile phase