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Dushyant Patel LC-MS/MS Method

S.K.P.C.P.E.R., Kherva 32 M .Pharm. Thesis

TABLE NO: 4.4 INSTRUMENTATION

TABLE NO: 4.5 REAGENTS AND CHEMICALS

Chemicals/Reagents Grade Manufacturer/ Supplier

Methanol LCMS grade Sigma-Aldrich, Germany

Water Milli Q Millipore, USA

Ammonium Formate For Molecular biology Sigma-Aldrich, Germany

4.2.2 PREPARATION OF SOLUTIONS:

a) Preparation of 5 mM Ammonium formate:

¾ Ammonium formate (315.30 mg) was weighed accurately and transferred in 1000 ml

glass bottle. Water (about 1000 ml) was added using graduated cylinder and sonicated to

dissolve.

¾ pH was adjusted to 3.0 with OPA.

b) Elution solution / Blank solution (Methanol 100 %):

c) Auto sampler rinsing solution (Methanol 100%):

d) Mobile phase preparation [Methanol: 5 mM Ammonium formate (80:20,% v/v)]

¾ In 1000 ml glass bottle, Methanol (800 ml) and 5mM Ammonium formate (200 ml) were

added using graduated cylinder. Mixed well. The solution was filtered on a 0.45µm

filter using solvent filtration apparatus by applying vacuum and sonicated it.

e) Preparation of diluent [Water : Methanol, (50: 50, % v/v)]:

¾ In 1000 ml glass bottle, water (500 ml) was taken using graduated cylinder. Methanol

(500 ml) was added using graduated cylinder. Mixed well.

Instrument Brand/Model Manufacturer/ Supplier

Auto sampler CTC PAL CTC PAL Ltd., UK

Solvent delivery module Accela pump Thermo Finnigan Ltd., UK

MS Detector

TSQ Quantum

ultra

Thermo Finnigan Ltd., UK

Vortexer Spinix Barnstead International, USA

Analytical balance Mettler Toledo Mettler Toledo India Pvt. Ltd., Mumbai

Solid Phase Extraction (SPE)

Unit

Waters/

Orochem

Waters, USA/Orochem, India

Freezer (-70°C) U725-86 New Brunswick Scientific co.inc., USA

Freezer (-20°C) 375 W Vest frost, UK

Refrigerator(2-8°C) RT34M Samsung India Electronics Ltd., India

Ultrasonic bath JAC 4020 Bio-sci instruments, India

Water purification system Milli-Q Millipore, USA

Vacuum Pump Millipore Millipore, India

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