Waters Forensic Toxicology Installation Standards Kit - Manual (Page 2)

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[ CARE AND USE MANUAL ]
Forensic Toxicology Installation Standards Kit
II. STORAGE AND STABILIT Y
The Waters Forensic Toxicology Installation Standards Kit is shipped
at ambient temperature, but should be refrigerated upon receipt.
The unopened kit is stable for the length of the expiration period if
it is stored refrigerated.
The kit should be allowed to equilibrate to room temperature
before use.
Diluted standard and prepared buffer are good for approximately
one week.
III. P REPARING T HE FORENSIC TOXICOLOGY
INSTALLATION STANDARD
The Forensic Toxicology Installation Standards is intended for use with
Xevo
® G2-S instruments use at a dilution of 1:20 using 18 MΩ cm
-1
deionized water.
Modify the dilution as required for other mass spectrometers.
Store both the concentrated and diluted standard in the refrigerator.
IV. BUFFER P REPARATION
Add the Ammonium Formate Concentrate (186007361-2) to
1.0 L of MΩ cm
-1
deionized water. This will make a final buffered
solution of 5 mM ammonium formate at pH 3.0.
When the intended application requires an accurate mobile phase
pH of the final solution must be verified before use. Use the
supplied formic acid (186007361-3) to adjust the actual pH as
required. As a guide, the addition of 100 µL of formic acid will
reduce the pH approximately 0.05 s.u.
V. EXAMPLE USING T HE TOXICOLOGY
INSTALLATION STANDARD
The example base peak intensity chromatogram (Figure 1) shows
typical retention time (Table 4) and relative responses using
positive ion electrospray with the LC conditions in Tables 2 and 3.
Table 2. Chromatographic Conditions
LC system: ACQUITY UPLC ® I Class
Column:
ACQUITY UPLC HSS C
18
, 1.8 µm,
2.1 x 150 mm (P/N 186003534)
Column temp: 50 °C
Sample temp: 10 °C
Mobile phase A:
5 mM aqueous ammonium formate
adjusted to pH 3.0 with formic acid
Mobile phase B: Acetonitrile with 0.1% formic acid
Wash solvent: 95:5 acetonitrile:water + 0.1% formic acid
Purge solvent:
5 mM aqueous ammonium formate
adjusted to pH 3.0 with formic acid
Seal wash: 80% water/20% methanol
Flow rate: 0.4 mL/min
Run time: 15 min
Injection volume: 10 µL
Table 3. LC Gradient
Time (min)
Flow Rate
(mL/min)
%A %B Curve
0.00 0.40 87.0 13.0 Initial
0.50 0.40 87.0 13.0 6
10.00 0.40 50.0 50.0 6
10.75 0.40 5.0 95.0 6
12.25 0.40 5.0 95.0 6
12.50 0.40 87.0 13.0 6
15.00 0.40 87.0 13.0 6
Table 4: Expected Analyte Retention Times for mobile
phase pH 3.0
Analyte
Retention Time
(minutes)
Neutral Mass
Nicotine 1.03 162.1157
Tiapride 1.69 328.1457
Scopolamine 2.19 303.1471
Buflomedil 4.30 307.1784
Milnacipran 4.99 246.1732
Trazodone 5.60 371.1513
Tripolidine ǂ 5.80 278.1783
Clozapine ǂ 6.20 326.1298
Tianeptine 6.90 436.1224
Perphenazine ǂ 8.80 403.1485
ǂ These analytes exhibit a shift in retention time as a function
of mobile phase pH at a rate of ~+0.1 min/+0.1 pH