March 26th, 2007

Chemistry Notes
Sciex Mass Spec

Page 1 of 13
Contents Page
Recommended Buffers 3
Probe Temperature 3
Compounds That Cause Sensitivity Suppression 3
Progesterone in Positive Ion ESI-MS 4
Effect of Phosphate Buffer 5
Progesterone in Positive Ion APCI (Effect of TEA contamination) 6
Taurocholic Acid by LC/MS (Example of Adducts and Wrong Polarity) 7
Adducts & Clusters formed in LC/MS 9
Choice of Correct Source: Erythromycin by + Ion TurboIonSpray 10
Choice of Correct Source: Erythromycin by + Ion Heated Nebulizer 11

Page 2 of 13
Turbo Ion Spray Guide
Recommended Buffers
• Ammonium formate or acetate (2-10 mM optimum, can see suppression effects over 20 mM).
• Acetic acid at 0.1 - 1.0 % Acetic
• Formic acid at 0.1% TFA (good for chromatography) at up to 0.1% for positive ion mode (signal supression sometimes with TFA).
• Do not use in negative ion mode.

Probe Temperature
• Distillation limits actual temperature experienced by Analyte ~ 150°C
• Minimum temperature set by solvent and flow rate.

Compounds That Cause Sensitivity Suppression

• Salts can interfere with ionization and can cluster to complicate spectrum (but also aid in identification).
• Strong bases or quaternary amines can interfere with positive mode analytes, e.g. Triethylamine (TEA)
• Acids - Sulfuric/Sulfonic acids and TFA interfere in negative mode experiments.
• Phosphate buffer and non-volatile ion pairing agents (e.g. SDS) can cause severe suppression and complex spectra.
• Dimerization ([2M+H]+) can occur at high concentration, leading to non-linearity during Quantitation.
• Dimer Signal at m/z = (MW*2)+1 can cause non-linearity at high concentrations.

Page 3 of 13
Progesterone in Positive Ion ESI-MS
[M+ Na]+

Page 4 of 13
Effect of Phosphate Buffer
–Q1 MCA (10 scans): from cck q1 -ve nano 8.78e7 cps
254.43
424.53
8e7

7e7

CCK dissolved in Phosphate Buffer

6e7

566.44
5e7
Intensity, cps

4e7

3e7
733.93
2e7
835.93

1e7
898.22 1000.42
1164.61
1142. 41

300 400 500 600 700 800 900 1000 1100

m/z, amu

–Q1 MCA (10 scans): from CCK PURIFIED Q1 SACN 16.7 4.44e7 cps
570.44

4.0e7

3.5e7
1142.03
Phosphate Buffer removed from CCK sample
3.0e7
Intensity, cps

2.5e7

2.0e7

1.5e7

292.82
377.02
1.0e7
439.03 879.23
497.03 700.63
798.63
5.0e6

300 400 500 600 700 800 900 1000 1100

m/z, amu Page 5 of 13
Progesterone in Positive Ion APCI (Effect of TEA contamination)

Page 6 of 13
Taurocholic Acid by LC/MS (Example of Adducts and Wrong Polarity)
.
21.9

9.0e6 +
[M + Na]
8.5e6
8.0e6
+
[M + NH ]
4
7.5e6 17.0 43.9
7.0e6 H 3C O
OH
6.5e6 CH3

6.0e6 CH3 NH
H
5.5e6
H H HO3S
5.0e6
HO
4.5e6
+ H
4.0e6 [M + H]
3.5e6 22.9 +
516.4
3.0e6
[M + K]
44.9
2.5e6 18.0 38.8

2.0e6 37.8
1.5e6
1.0e6
5.0e5
0.0
515 520 525 530 535 540 545 550 555 560 565 570 575
m/z, amu
Page 7 of 13
Adducts & Clusters formed in LC/MS
Covalent*/
Cluster/Adduct Ion Cause Ionic APCI ESI +/- m/z of Ion

Lithium Lithium Salts Ionic 3 3 + [M + H + 6]+

Ammonia Ammonia/
[M + NH4]+ NH4OH Covalent 3 3 + [M + H + 17]+
Water Water/
[M + H3O] +
Acids Covalent 3 3 + [M + H + 18]+
Sodium
[M + Na]+ Sodium salts Ionic 3 3 + [M + H + 22]+
Methanol Methanol in [M + H + 32]+
[M + H + CH3OH] +
Solvent Covalent 3 3 +/- [M - H + 32]-
Chloride
[M + Cl]- Chlorinated solvent Ionic 3 6 - [M + 35 (& 37)]-
Potassium
[M + K]+ Potassium salts Ionic 3 3 + [M + H + 38]+
Acetonitrile Acetonitrile in [M + H + 41]+
[M + H + CH3CN)+ solvent Covalent 3 3 +/- [M - H + 41]-
Formic Acid
[M - H + HCO2H]- Formic Acid Covalent 3 3 - [M - H + 46]-
Acetic Acid
[M - H + CH3CO2H]- Acetic Acid Covalent 3 3 - [M - H + 60]-
TFA
[M - H + CF3CO2H]- TFA Covalent 3 3 - [M- H + 114]-
* Covalent adducts can often be broken up by using a higher DP voltage.

Page 8 of 13
Negative Ion TurboIonSpray with NaCl-containing eluents.
–Q1: 2.53 min (50 scans) from Grundlinie 10mM NaCl 1.24e5 cps
269

1.0e5
Intensity, cps

151 211
5.0e4
327
445
130 235
177 293 353 385 411
0.0e0

150 200 250 300 350 400

m/z, amu

–Q1: 1.17 min (26 scans) from Grundlinie 5mM NaCl+NH4ac 4.41e4 cps

1.0e5
Intensity, cps

5.0e4
141
269
151 211 227 255
177 293
317 329 341 351 375 411 443
0.0e0

150 200 250 300 350 400

m/z, amu

–Q1: 1.08 min (25 scans) from Grundlinie 10mM NH4ac, subtracted (scans 4 to 24) 6.65e3 cps

1.0e5
Intensity, cps

5.0e4

141 161 189 217 233 265 277 299 329 372 413 432
150 200 250 300 350 400
m/z, amu

Page 9 of 13
Choice of Correct Source: Erythromycin by + Ion TurboIonSpray

Page 10 of 13
Choice of Correct Source: Erythromycin by + Ion Heated Nebulizer

Page 11 of 13
Page 12 of 13
Page 13 of 13