MOOC: Validation of liquid chromatography mass spectrometry (LC-MS) methods (analytical chemistry) course

Self test 10.4

1.

An analyst has developed a LC/MS method for determining pesticides in tomatoes. One important aspect for your method is that each analyte is analyzed in a fixed time window.

Time window

Analyte

Retention time (min)

m/z of precursor ion

m/z of product ion

1

oxamyl

7.8

242

185

2

thiamethoxam

9.1

292

211

carbendazim

8.5

192

160

3

imidacloprid

10.3

256

209

thiabendazole

9.6

202

175

4

acetamiprid

11.2

223

126

vamidothion

10.9

310

146

5

thiacloprid

12.0

253

126

6

spiroxamine

15.2

298

144

imazalil

13.9

297

201

7

mepanipyrim

17.3

224

183

fluquinconazole

17.1

376

349

methiocarb

16.5

226

169

8

buprofezin

18.8

306

201

hexythiazox

19.3

353

228

9

etofenprox

20.9

359

189

Therefore it is important to verify if the method is robust enough from the retention times point of view (i.e. if the retention times are stable enough). Analyst uses isocratic elution with eluent composition 30% (v/v) of 1 mM acetic acid buffer with pH 4.5 and 70% (v/v) of acetonitrile. Analyst wants to study the effect of buffer concentration (0.9 mM to 1.1 mM)|pH (4.0 to 5.0) and organic modifier content (68% to 72%). Please indicate which experiments need to be carried out to evaluate the robustness with full factorial design.

% MeCN pH Buffer concentration (mM)
68 4.0
68 1.1
5.0 0.9
68 5.0
72 0.9
72 4.0
5.0 0.9
5.0 1.1

2.

Imagine we want to perform a full factorial design with four factors that influence our response. How many experiments need to be performed?

3.

The analyst is evaluating robustness with full factorial design. Most of all she is concerned about limit of detection. From previous knowledge she knows that LoD in case of her method is mostly influenced by ionization source parameters. Therefore she determined LoD dependence on nebulizer gas pressure and drying gas temperature (data given below).

Factor A

Nebulizer gas pressure (psi)

Factor B

Drying gas temp. (°C)

LoD (µg/L)

55

360

3.2

55

340

3.8

45

360

2.8

45

340

3.7

Calculate how large is the effect of the nebulizer gas pressure|on the drying gas temperature and on the interaction of drying gas temperature* nebulizer gas pressure on the LoD!

Please use dots (.) instead of commas (|) for separating the decimals and present the answer with 2 decimals.

The effect of the factor A (nebulizer gas pressure) on LOD is   µg/L.

The effect of the factor B (drying gas temperature) on LOD is   µg/L.

The effect of the interaction of factors A and B (drying gas temperature*nebulizer gas pressure) on LOD is   µg/L.

Create an order of significants of effects and interactions from most impactful (top) to least impactful (bottom).

1. (most impactful) 

2. 

3. (least impactful) 

4.

contour plot

Which of the following statements about the interaction plot are true :

5.

contour plot

Which of the following statements about the contour plot above are true: