Table 4.
Analytical methods for amatoxins detection.
| Ref. | Matrix | Separation | Detection | Qualitative/Quantitative | LOD | LOQ | Linearity | Extraction Recovery | Additional Analytical Information |
|---|---|---|---|---|---|---|---|---|---|
| [119] | Rabbit serum | - | RIA | Qualitative | α-: 50 pg | NA | NA | NA | - |
| [70] | Pure substances | TLC | - | Qualitative | α-: 50 µg | NA | NA | NA | - |
| [120] | Mushrooms | HPTLC | Spectrophotometry | Quantitative | 50 ng deposit | NC | NC | NC | - |
| [121] | Serum, urine, duodenal fluid, gastric juice, mushrooms | - | RIA | Quantitative | 3 µg/L | NC | 3.3–100 µg/L | NC | - |
| [122] | Serum, urine, stomach washings | HPLC | UV (280 nm) | Quantitative | 10 µg/L | NC | 20–500 µg/L | 110% | Column: (250 mm × 4.6 mm) 5 µm Ultrasphere ODS C18; Flow rate: 1 mL/min; Mobile phase: 0.02 M ammonium acetate/ACN (88/12; v/v) pH 5; RT α-: 12.1 min, β-: 7.4 min |
| [123] | Serum, urine, mushrooms | HPLC | UV (302 nm) | Quantitative | 10 ng | NC | 0.5–20 mg/L | α-: 81.1–98.1% β-: 80.6–97.3% |
Column: (125 mm × 4.0 mm) 5 µm Lichrosorb RP-18; Flow rate: 1 mL/min; Mobile phase: ACN (A), 0.01 M acetic acid-ammonium acetate buffer pH 5 (B); RT α-: 14.9 min, β-: 9.1 min |
| [124] | Plasma, urine | - | RIA | Quantitative | 0.1 µg/L plasma 1 µg/L urines |
NC | 0.1–20 µg/L plasma; 1–100 µg/L urines | 101.3% plasma 110% urine |
- |
| [125] | Serum, urine | HPLC | Amperometry (Reference electrode: Ag/AgCl; Working potential: 600 mV) |
Quantitative in serum; Qualitative in urine | α-: 40 pg on column β-: 80 pg on column |
NC | 1–1000 µg/L | α-: 53–65% β-: 36% |
Column: (250 mm × 4.6 mm) 5 µm Spherisorb ODS2 - (250 mm × 4.6 mm) 5 µm Hypersil WP300 Butyl; Flow rate: 1 mL/min; Mobile phase: 0.02 M ammonium acetate/ACN (92:8; v/v) 0.5 mM EDTA pH 5; RT α-: 16.5 min, β-: 12.0 min |
| [126] | Plasma | HPLC | UV (303 nm) | Quantitative for α-amanitin | 9.74 µg/L | 10 µg/L | 10–100 µg/L | 67.3–105.56% | - |
| [127] | Plasma | HPLC | Amperometry/EC (Reference electrode: Ag/AgCl; Working potential: 350 mV) |
Quantitative for α-amanitin | 2 µg/L | NC | 3–200 µg/L | 80–82.5% | Column: (150 mm × 4.6 mm) 5 µm PLRP-S 100 Å; Flow rate: 0.5 mL/min; Mobile phase: 0.05 M phosphate buffer—ACN (91/9; v/v) pH 9.5 |
| [128,129] | Mushrooms | HPLC | UV (214, 295 nm) | Quantitative | 10 µg/L = 0.5 ng/g mushrooms | NC | NC | NC | Column: (250 mm × 4.6 mm) 5 µm Ultrasphere ODS; Flow rate: 1 mL/min; Mobile phase: 0.02 M aqueous ammonium acetate/ACN (90/10; v/v A) (76/24; v/v B) |
| [63] | Urine, mushrooms | Electrophoresis | DAD: 190–350 nm | Quantitative | 1000 µg/L | NC | 1–1000 mg/L | NC | Capillary length: 36 cm (50 µm); T separation: 25°C; Buffer: 100 mM phosphate (pH 2.4) |
| [130] | Urine | HPLC | Coulometry (Full scale range 50 µA until 12.5 min, 20 µA up to 20 min) |
Quantitative for α-amanitin | 2 µg/L | 10 µg/L | 10–200 µg/L | 77–80.4% | Column: (250 mm × 4.6 mm) Supelcosil LC 18; Flow rate: 1 mL/min; Mobile phase: 0.005 M bisodic phosphate aqueous solution pH 7.2 and ACN (90/10; v/v); Electrode: graphite |
| [131] | Plasma, urine | HPLC | ESI-UV-MS (UV: 302 nm) (SIM mode (+): α- 919, 920, 921 m/z; β- 920, 921, 922 m/z) |
Quantitative | 2.5 µg/L | 5.0 µg/L | 5–75 µg/L | α-: 49.1–62.5% β-: 52.1–57.5% |
Column: (100 mm × 2.1 mm) 3 µm HP ODS Hypersil RP-18; Flow rate: gradient; Mobile phase: MeOH-0.01 M ammonium acetate pH 5 (10/90; v/v A) (70/30 v/v B) |
| [132] | Serum, urine | ELISA | - | Quantitative for β-amanitin | 0.08 µg/L | NC | 0.080–2 µg/L | NC | - |
| [133] | Mushrooms | HPLC | HILIC-ESI-MS/MS (ion trap) (scan range: 600–930 m/z) |
Quantitative | 20 ng/g | α-: 26.8 ng/g β-: 33.3 ng/g |
20–500 µg/L | 63–75% | Column: (250 mm × 2.0 mm) 5 µm 80 Å TSK-Gel Amide 80; Flow rate: 0.2 mL/min; Mobile phase: 2 mM ammonium formate + 5mM HCOOH (A), ACN (B), MeOH (C); RT: α- ≈ 7.18 min, β- ≈ 8.94 min |
| [134] | Serum, liver | HPLC | ESI-MS/MS/MS (ion trap) (α- 941 to 746 (CE 40%) m/z; Full-scan of product ions of m/z 746 (CE 25%)) |
Quantitative for α-amanitin | 0.26 ng/g (serum) 0.5 ng/g (liver) |
NC | 1–50 µg/L | 95% (serum) 98% (liver) |
Column: (100 mm × 4.6 mm) Synergi RP-Polar; Flow rate: 0.5 mL/min; Mobile phase: 0.01 M ammonium acetate in H2O 0.1% HCOOH (A), 0.01 M ammonium acetate in MeOH 0.1% HCOOH (B); RT: α-: 4.5 min |
| [135] | Urine | Electrophoresis | DAD (214 nm) | Quantitative | 2.5 µg/L | 5 µg/L | 5 - 100 µg/L | NC | Capillary length: 48 cm (75 µm); T separation: 25 °C |
| [136] | Plasma | HPLC | ESI-MS/MS (ion trap) (SIM mode: α- 919–921 m/z; β- 920–922 m/z) |
Quantitative | 0.5 µg/L | NC | 10–500 µg/L | 77–79% | Column: (150 mm × 2.0 mm) Capcell Pak C18 UG120; Flow rate: 0.2 mL/min; Mobile phase: H2O 0.1% HCOOH (A), ACN 0.1% HCOOH (B); RT: α-: 19.0 min, β-: 20.1 min |
| [137] | Mushrooms | HPLC | ESI-TOF-MS (Full-scan: 100–1000 m/z) |
Quantitative | 30 ng/g | NC | 100–1000 ng/g | 53.1–69.6% | Column: (150 mm × 2.0 mm) 3 µm TSK-gel Amide-80; Flow rate: 1 mL/min; Mobile phase: ACN (A), 15% MeOH in 10 mM ammonium acetate (B) |
| [11] | Serum, urine | UPLC | ESI-MS/MS (triple Q) (α- 919.6 to 919.6 (20 eV) m/z; β-: 920.6 to 920.6 (20 eV) m/z) |
Quantitative | 0.5–1.5 µg/L | NC | 2–420 µg/L | 91.3–110% | Column: (100 mm × 2.1 mm) 1.7 µm ACQUITY BEH Shield RP18; Flow rate: 0.4 mL/min; Mobile phase: H2O 0.1% HCOOH (A), MeOH (B); RT: α-: 2.23 min, β-: 2.49 min |
| [138] | Urine | MALDI | ESI-TOF-MS-MS | Quantitative | 0.5 µg/L | NC | 10–500 µg/L | 60–80% | - |
| [139] | Urine, liver | UPLC | ESI-MS/MS (triple Q) (α-: 919.48 to 259.13 (44 eV)/919.48 to 901.53 (28 eV) m/z; β-: 920.48 to 259.13 (42 eV)/920.48 to 902.44 (26 eV) m/z) |
Quantitative | 0.20 µg/L (urine) 10 ng/g (liver) |
0.46–0.57 µg/L (urine) 12.3–14.7 ng/g (liver) |
10–200 µg/L (et ng/g) | 90.4–105.0% (urine) 90.2– 12.9% (liver) |
Column: (100 mm × 2.1 mm) 1.8 µm ACQUITY HSS T3; Flow rate: 0.5 mL/min; Mobile phase: 0.02 M ammonium acetate pH 5 (A), ACN (B); RT: α-: 5.73 min, β-: 5.27 min |
| [140] | Urine | UPLC | (-) ESI-HR/MS/MS (orbitrap) (SIM mode: α-: 917.3458 m/z; β-: 918.3298 m/z) |
Quantitative for α-amanitin | 1 µg/L | 1 µg/L | 1–100 µg/L | 64–102% | Column: (150 mm × 2.1 mm) 2.6 µm TF Accucore PhenylHexyl; Mobile phase: 10 mM ammonium acetate in H2O 0.01% HCOOH pH 5 (A), ACN 0.1% HCOOH (B), 2-propanol/ACN (1:1; v/v) (C); RT: α-: 8.23 min, β-: 7.61 min |
| [141] | Urine | UPLC | HR/MS/MS (orbitrap) (SIM mode: α-: 919.3614 m/z; β-: 920.3455 m/z) |
Quantitative | α-: 0.25 µg/L β-: 0.5 µg/L |
α-: 0.5 µg/L β-: 0.75 µg/L |
1–100 µg/L | 88.4–93.4% | Column: (100 mm × 2.1 mm) 2.6 µm Accucore C18; Flow rate: 0.4 mL/min; Mobile phase: 10 mM ammonium acetate buffer 0.1% HCOOH (A), ACN 0.1% HCOOH (B); RT: α-: 1.9 min, β-: 1.7 min |
| [142] | Mushrooms | HPLC | DAD (303 nm) | Quantitative | 2 ng/g | NC | NC | NC | Column: (150 mm × 4.6 mm) 5 µm C18; Flow rate: 1 mL/min; Mobile phase: 0.05 M ammonium acetate pH 5.5 with HCOOH/ACN (90:10; v/v) |
| [143] | Urine | UPLC | ESI-TOF/MS (Full-scan 50–1000 m/z) |
Quantitative | 1 µg/L | NC | 1–1000 µg/L | 86–98% | Column: (100 mm × 2.1 mm) 2.2 µm Acclaim RS 120, C18; Flow rate: 0.2 mL/min; Mobile phase: H2O/ACN (99/1; v/v) 2mM ammonium formate, 0.1% HCOOH (A), ACN/H2O (99/1; v/v) 2mM ammonium formate, 0.1% HCOOH (B); RT: α-: 6.05 min, β-: 6.08 min |
| [144] | Rat liver and kidney Serum |
HPLC | DAD-EC (UV: 305 nm) |
Quantitative for α-amanitin | UV: 110 ng/g (liver) 160 ng/g (kidney) EC: 70 ng/g (liver) 40 ng/g (kidney) |
UV: 330 ng/g (liver) 500 ng/g (kidney) EC: 210 ng/g (liver) 110 ng/g (kidney) |
UV: 330–10000 µg/L (liver) 500–10000 µg/L (kidney) EC: 210–10000 µg/L (liver) 110–10000 µg/L (kidney) |
UV: 99.4% (liver) 100% (kidney) EC: 98.8% (liver) 99.7% (kidney) |
Column: (250 mm × 4.6 mm) 5 µm Spherisorb RP-18 ODS2; Flow rate: 1 mL/min; Mobile phase: 20% MeOH in 50 mM citric acid, 0.46 mM octanessulfonic acid pH 5.5 with 10 M NaOH |
| [145] | Serum, urine | UPLC | ESI-MS/MS (triple Q) (α-: 919.5 to 259.1 (42 eV)/919.5 to 86.0 (68 eV) m/z; β-: 920.5 to 259.1 (42 eV)/920.5 to 86.0 (71 eV) m/z) |
Quantitative | 0.5–1 ng/g | 1–2.5 ng/g | 1–100 µg/L | 80.7–88.6% | Column: (100 mm × 2.1) 1.6 µm; Flow rate: 0.2 mL/min; Mobile phase: 0.2% HCOOH in H2O (A), 0.2% HCOOH in MeOH (B); RT α-: 4.72 min, β-: 4.96 min |
| [146] | Food with mushrooms | HPLC | (-) ESI-MS/MS (triple Q) (α-: 917.4 to 205.1/917.4 to 257.1 m/z; β-: 918.4 to 205.1/918.4 to 257.1 m/z) |
Quantitative | 5 ng/g | 10 ng/g | 10–2000 ng/g | 77.6–90.4% | Column: (150 mm × 3.0 mm) 2.5 µm XBridge™ BEH C18; Flow rate: 0.3 mL/min; Mobile phase: MeOH (A), 0.03% ammonia solution in H2O pH 10.5 (B) |
| [147] | Rat plasma | HPLC | (+) ESI-MS/MS (triple Q) (MRM: 919.45 to 259.20 (47 eV); 919.45 to 901.45 (26 eV); 919.45 to 86.15 (50 eV) m/z) |
Quantitative for α-amanitin | 3.0 µg/L | 8.5 µg/L | 10–1500 µg/L | 85–115% | Column: (100 mm × 2.1 mm) 5 µm Hypersil GOLD C18; Flow rate: 0.2 mL/min; Mobile phase: 0.02 mol/L ammonium acetate, 0.1% HCOOH (A), ACN (B); RT: 4.86 min |
| [148] | Rat plasma and urine | HPLC | PDA-MS/MS/MS (IT-TOF) (PDA scan: 190–400 nm; Full-scan: 700–1000 m/z; Multiple stage fragmentation: 100–900 m/z for MS2, 50–900 m/z for MS3) |
Qualitative | NC | NA | NA | NA | Column: (100 mm × 2.1 mm) 3µm Inertsil ODS-3; Flow rate: 0.2 mL/min; Mobile Phase: 20 mM ammonium acetate, 0.1% HCOOH (A), ACN (B); RT α-: 11.05 min, β-: 10.20 min |
| [149] | Urine | HPLC | ESI-MS/MS (triple Q) (α-: 919.3 to 338.9 m/z; 15N10- α-: 929.3 to 911.4 m/z, β-: 920.3 to 644.3 m/z) |
Quantitative with 15N10-α-amanitin | α-: 0.458 µg/L β-: 0.930 µg/L |
NC | α-:1–200 µg/L β-: 2.5–200 µg/L |
α-: 97.8% β-: 71.1% |
Column: (50 mm × 2.1 mm) 1.7 µm Acquity BEH HILIC; Flow rate: gradient; Mobile phase: 10 mM ammonium formate in ACN (25/75; v/v) 1% HCOOH (A), 10 mM ammonium formate in ACN (10/90; v/v) 0.2% HCOOH (B) |
| [56] | Standard solution | - | PSI-HR-MS/MS (α-: 919.3610 to 86.0606 m/z; β-: 920.3405 to 86.0606 m/z) |
Qualitative | NA | NA | NA | NA | - |
| [150] | Mushrooms | - | LFIA | Qualitative | α-: 10 µg/L β-: 2000 µg/L γ-: 10 µg/L |
NA | NA | NA | - |
NA: not applicable; LOD: limit of detection; LOQ: limit of quantification; NC: not communicated; RT: retention time; DAD: diode array detection; EC: electrochemical.