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Calibration Curve for Cyanuric acid Extraction (20-2000ng/mL) from Strata Melamine on Luna HILIC

Sample Preparation Details

Condition 1 :3 mL Methanol at 1.0 mL/min
Condition 2 :3 mL Water at 1.0 mL/min
Load :Load 3.50 mL Sample diluted in Acetonitrile:Water
Wash 1 :1 mL Acetonitrile:Water (50/50); 2 aliquots of 500 uL at 1.0 mL/min
Wash 2 :0.5 mL Methanol:Water (50/50); 1 aliquot of 500 uL at 1.0 mL/min
Wash 3 :2 at 1.0
Dry :2 mins at 10" of Hg
Elution 1 :0.5 mL Methanol at 1.0 mL/min
Elution 2 :1.0 mL 5% Ammonium Hydroxide in Methanol (aliquots of 0.5 mL) at 1 mL/min
Evaporate: :Evaporate to dryness
Reconstitute :1 mL 90/10 : ACN/100mM ammonium formate, pH 3.2
Notes :Protein Precipitation: Weigh 0.2 mg (200mg) of Baby formula (Enfamil) in a 15 ml plastic centrifuge tube, add 1 ml Lukewarm water to make a homogeneous mix, vortex and add 100uL of 0.2N HCl followed by addition of 50uL of each internal standard (at 15.0 ug/mL). Vortex 3-5 secs. Add 3 ml of ACN, vortex and centrifuge at 6000Xg for 10 mins and then collect the supernatant to carry out SPE in the subsequent step.

Application Detail (App ID: 18468)

Part No:00D-4449-B0
Dimensions:100 x 2 mm ID
Elution Type:Isocratic
Elution A:90/10 ACN/100 mM Ammonium Formate, pH 3.2
Gradient Profile:
Step No. Time (min) Pct A
1 3.5 100

Flow Rate:0.4 mL/min
Col. Temp: ambient
Detection: Mass Spectrometer (MS) @ (22°C)
Detector Info: * MS in -ve ionization till 1.7 mins then to +ve till 3.5 mins
Note:LC/MS/MS Condition:
Column: Luna 3µm HILIC, 100x2.0mm; Flow rate: 0.4ml/min
Mobile Phase: Isocratic; 90/10 ACN/100mM NH4 Formate pH 3.2; MS Detector: API 3000 LC/MS/MS with ESI (TurboIonSpray®) (www.Appliedbiosystems.com), Switch from -ve Ion Mode (for cyanuric) to Positive Ion (for Melamine);
MRM; heater gas flow 7000 cc/min; heater temperature 450 °C

Cyanuric acid QC - 200 ng
% Accuracy 100.1%
% CV 17%


Calibration Curve of Extracted Samples

conc. (ng) % Accuracy
20 104%
40 98%
200 95%
400 103%
2000 99.91%

Analyte Details

Analyte Detail not available

1
Cyanuric acid
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2
Cyanuric-13C3
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