| RP-SPE |
porcine insulin, novolog, apidra,
lantus DesB(30–32) metabolite,
humalog and human insulin,
GH-RH(1–29), LH-RH, synacthen, CJC-1295, LongR3-IGF-1, and IFG-1 |
preconditioned
solution: ACN (2 mL), water (2 mL); load solution: 1–10 mL of urine fortified with ISs (500 pg/mL)
and 10% of ACN); rinsing solution: water; eluting solution: ACN/water (80:20, v/v)54
|
| |
GHRPs, desmopressin, vasopressin, felypressin,
terlipressin,
leuprolide, LHRH, ibutamoren, anamorelin |
preconditioned
solution: MeOH (1 mL), water (1 mL); load solution:
7 mL of urine, 40 μL of IS (100 ng/mL), pH 6.5–7.5, 100 μL of PBS; rinsing solution: water (1 mL), 20% ACN in
water (1 mL); eluting solution: 1 mL of 95/5 (75% ACN in water/FA)55
|
| |
GHRPs, desmopressin, vasopressin, and leuprolide |
preconditioned solution: MeOH (12 mL, 3×), water (12 mL,
3×); load solution: peptide (50 mg/mL) in water with 0.5% TFA; rinsing solution: water with 0.5% TFA (12
mL); eluting solution: gradient of 5–50% ACN with 0.5% TFA62,63
|
| RP-SPE/IE-SPE |
GHRPs and opioid peptides |
preconditioned solution: MeOH (2 mL), water (2 mL); load solution:
plasma containing 10 ng mL–1 each of the peptides;
rinsing solution: ACN/water; eluting solution: ACN (10–90%)/ water with 2% FA32
|
| |
GHRPs, anamorelin,
leuprolide, LHRH, triptorelin, [deamino-Cys1-Val4-d-Arg8]-vasopressin (ISTD) |
preconditioned
solution: MeOH (1 mL), water (1 mL); load solution:
1 mL of urine; rinsing solution: 1 mL 5% of ammonium hydroxide and
1 mL 20% ACN; eluting solution: 0.5 mL of NH3/FA 10%/MeOH 8/12/80 v/v50
|
| |
GHRPs, anamorelin, and [deamino-Cys1-Val4-d-Arg8]-vasopressin (ISTD) |
preconditioned solution: MeOH (1 mL), water (1 mL); load solution:
2 mL of sample (HRPs), 40 μL IS (2 μg/mL diamino-Cys1Val4d-Arg8-vasopressin), 200
μL 0.8 M PBS, pH 7; rinsing solution: 1 mL water and 0.5 mL
of MeOH; eluting solution: ACN/water with 2% FA 1:3 v/v79
|
| microextraction RP-SPE |
GHRPs, desmopressin, vasopressin, felypressin, terlipressin,
leuprolide, LHRH, ibutamoren, and anamorelin |
preconditioned
solution: MeOH (0.2 mL), water (0.2 mL); load
solution: 0,75 mL of urine, 15 μL of IS (100 ng/mL), pH 6.5–7.5, 50 μL of PBS; rinsing solution:
water (0.2 mL), 20% ACN in water (0.2 mL); eluting solution: 25 μL
of 95/5 (75% ACN in water/FA) (2×)55
|
| enzymatic digestion |
GHRPs, TB-500 and desmopressin |
incubation: 24
h at 37 °C; enzyme/protein ratio: 1:20, 1:50, and 1:100 (w/w); peptide concentration: 0.2 mg/mL; buffer
solutions: 50 mM Tris-HCl, pH 7.0 (rhAPN); 50 mM MES, pH 6.0, 5 mM
CaCl2 (rhCPM); 0.2 M Na2HPO4/0.2
M NaH2PO4 2/7 (v/v) (Leu-AP); 50 mm Tris-HCl, pH 7.65, 0.1 M NaCl (rhCPB and CPB);
25 mM Tris-HCl, pH 8.85, 1 mM EDTA (endopeptidase Lys C)56
|
| |
GHRPs, sermorelin, IGF-1, hCG |
incubation: 37 °C for 16 h; enzyme/protein ratio: 1:200 (w/w); peptide concentration: 0.2 mg/mL; buffer
solutions ammonium bicarbonate, 100 μL of 50 mM acetic acid,
pH 8 (2.5 μg sequencing grade trypsin)80
|
| liquid–liquid extraction |
GHRPs, anamorelin, and [deamino-Cys1-Val4-d-Arg8]-vasopressin (ISTD) |
sample
solution: 2 mL of sample, 200 μL IS (2 μg/mL
diamino- Cys1Val4d-Arg8-vasopressin),
200 μL ACN, 200 μL carbonate buffer, pH 12; extraction
solution: ethyl acetate, shaking, 30 min79
|
| immunoaffinity |
porcine insulin, novolog, apidra, lantus DesB(30–32) metabolite, humalog and human insulin,
synacthen, LH-RH, GH-RH(1–29), CJC-1295, longR3-IGF-1 and IFG-1 |
samples dissolved in PBS, pH 7.4, and mixed with primary
antibodies (1–3 μg of
IgG), paramagnetic beads
solution (40 μL of antimouse, 40 μL of antirabbit and
20 μL of protein A suspension), incubation for 1–3 h at 22 °C54
|
