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. 2022 Mar 19;31(8):935–956. doi: 10.1007/s10068-022-01056-5

Table 3.

Various analytical methods for the determination of phenolics from common buckwheat sprouts

Instrument typea Stationary phaseb Mobile phasec Flow rate; injection Identification and/or quantificationd Phenolics analytee References
UFLC C18 RP (5 μm, 250 × 4.6 mm)

A: 2% acetic acid in water

B: 2% acetic acid in 45% acetonitrile

1 mL/min; 10 μL

355 nm

External standard (RT)

External standard calibration

Flavonoid Lim et al. (2012)
HPLC–UV C18 RP (3 μm, 250 × 2 mm); 45 °C

A: acetonitrile/water/formic acid (6:89:5)

B: acetonitrile/water/formic acid (80:15:5)

0.2 mL/min

350 nm

External standard (RT)

External standard calibration

Flavonoid Horbowicz et al. (2013)
CE–UV Silica NP (75 μm); 25 °C 50 mM boric acid buffer containing 0.1 M SDS (pH 9.0) Pressure injection (20 psi × 10 s)

280 nm

External standard (RT)

External standard calibration

Flavonoid Koyama et al. (2013)
HPLC–UV C18 RP (150 × 4.6 mm); 40 °C

A: 0.1% formic acid in water

B: 0.1% formic acid in acetonitrile

0.8 mL/min; 10 μL

280 nm

External standard (RT)

External standard calibration

HPLC–MS C18 RP (150 × 4.6 mm); 40 °C

A: 0.1% formic acid in water

B: 0.1% formic acid in acetonitrile

0.8 mL/min; 10 μL

ESI positive mode

Scanning (100–1000 m/z)

External standard (RT) + MS spectra

HPLC–UV C18 RP (150 × 4.6 mm); 40 °C

A: 0.1% formic acid in water

B: 0.1% formic acid in acetonitrile

0.8 mL/min; 10 μL

280 nm

External standard (RT)

Rutin standard calibration

Flavonoid Nakamura et al. (2013)
HPLC–UV C18 RP (250 × 4.6 mm) Acetonitrile/0.1% formic acid (16:84) 1 mL/min

370 nm

External standard (RT)

External standard calibration

Flavonoid Tsurunaga et al. (2013)
HPLC–UV C18 RP (5 μm, 250 × 4.6 mm); 40 °C

A: methanol/water/acetic acid (5:92.5:2.5)

B: methanol/water/acetic acid (95:2.5:2.5)

1 mL/min; 10 μL

350 nm

External standard (RT)

External standard calibration

Flavonoid and phenolic acid Arasu et al. (2014)
HPLC–UV C18 RP (5 μm, 250 × 4.6 mm); 40 °C

A: methanol/water/acetic acid (5:92.5:2.5)

B: methanol/water/acetic acid (95:2.5:2.5)

1 mL/min; 10 μL

350 nm

External standard (RT)

External standard calibration

Flavonoid and phenolic acid Kim et al. (2014b)
HPLC–UV C18 RP (5 μm, 250 × 4.6 mm); 40 °C

A: methanol/water/acetic acid (5:92.5:2.5)

B: methanol/water/acetic acid (95:2.5:2.5)

1 mL/min; 10 μL

350 nm

External standard (RT)

External standard calibration

Flavonoid and phenolic acid Lee et al. (2014)
HPLC–UV C18 RP (5 μm, 250 × 4.6 mm); 50 °C

A: 0.2% acetic acid in water

B: 0.2% acetic acid in methanol

5 μL

350 nm

External standard (RT)

External standard calibration

Flavonoid Ren and Sun (2014)
HPLC–DAD C18 RP (3 μm, 250 × 2 mm); 45 °C

A: acetonitrile/water/formic acid (6:89:5)

B: acetonitrile/water/formic acid (80:15:5)

0.2 mL/min

350 nm (flavonoid); 520 nm (anthocyanin)

External standard (RT)

External standard calibration

Flavonoid Wiczkowski et al. (2014)
HPLC–MS C18 RP (3 μm, 250 × 2 mm); 45 °C

A: acetonitrile/water/formic acid (6:89:5)

B: acetonitrile/water/formic acid (80:15:5)

0.2 mL/min

ESI positive mode

External standard (RT) + MS spectra

HPLC–CoulArray–ECD C18 RP (3 μm, 250 × 2 mm); 45 °C Acetonitrile/water/phosphoric acid (9.99:89.99:0.02) 0.2 mL/min

 + 800–950 mV

External standard (RT)

External standard calibration

HPLC–DAD C18 RP (3 μm, 250 × 2 mm); 45 °C

A: acetonitrile/water/formic acid (6:89:5)

B: acetonitrile/water/formic acid (80:15:5)

0.2 mL/min

350 nm (flavonoid); 520 nm (anthocyanin)

External standard (RT)

External standard calibration

Flavonoid Horbowicz et al. (2015)
HPLC–MS C18 RP (3 μm, 250 × 2 mm); 45 °C

A: acetonitrile/water/formic acid (6:89:5)

B: acetonitrile/water/formic acid (80:15:5)

0.2 mL/min

ESI positive mode

External standard (RT) + MS spectra

HPLC–PDA C18 RP (5 μm, 250 × 4.6 mm); 25 °C

A: 0.1% formic acid in water

B: 0.1% formic acid in acetonitrile

0.8 mL/min; 20 μL

350 nm

External standard (RT)

Flavonoid Nam et al. (2015)
HPLC–Q–TOF/MS C18 RP (5 μm, 250 × 4.6 mm); 40 °C

A: 0.1% formic acid in water

B: 0.1% formic acid in acetonitrile

0.8 mL/min; 20 μL

ESI positive and negative mode

Scanning (50 – 1700 m/z)

External standard (RT) + MS spectra

HPLC–UV C18 RP (5 μm, 250 × 4.6 mm); 30 °C

1) C-glycosyl flavones analysis:

A: 0.1% formic acid in water

B: methanol

2) Other flavonoid analysis:

A: 0.15% phosphoric acid in water

B: acetonitrile

3) Phenolic acid analysis:

A: 0.15% phosphoric acid in water

B: methanol

1 mL/min (flavonoid), 0.7 mL/min (phenolic acid); 20 μL

355 nm (flavonoid); 270 nm (phenolic acid)

External standard (RT)

Flavonoid and phenolic acid Zhang et al. (2015)
HPLC–MS C18 RP (5 μm, 250 × 4.6 mm); 30 °C

1) C-glycosyl flavones analysis:

A: 0.1% formic acid in water

B: methanol

2) Other flavonoid analysis:

A: 0.15% phosphoric acid in water

B: acetonitrile

3) Phenolic acid analysis:

A: 0.15% phosphoric acid in water

B: methanol

1 mL/min (flavonoid), 0.7 mL/min (phenolic acid); 20 μL

ESI negative mode

Scanning (50 – 1000 m/z)

External standard (RT) + MS spectra

External standard calibration

HPLC–UV C18 RP (3 μm, 250 × 2 mm); 45 °C

A: acetonitrile/water/formic acid (6:89:5)

B: acetonitrile/water/formic acid (80:15:5)

0.2 mL/min

350 nm (flavonoid); 520 nm (anthocyanin)

External standard (RT)

External standard calibration

Flavonoid Dębski et al. (2016)
HPLC–UV C18 RP (150 × 4.6 mm); 40 °C

A: 0.1% formic acid in water

B: 0.1% formic acid in acetonitrile

0.8 mL/min; 10 μL

280 nm

External standard (RT)

Rutin standard calibration

Flavonoid Nakamura et al. (2016)
HPLC–MS C18 RP (2.6 μm, 100 × 2 mm); 25 °C

A: 0.1% formic acid in water

B: acetonitrile

0.3 mL/min; 10 μL

ESI negative mode

External standard (RT) + MS spectra

External standard calibration

Flavonoid and phenolic acid Terpinc et al. (2016)
HPLC–DAD C18 RP (3 μm, 250 × 2 mm); 45 °C

A: acetonitrile/water/formic acid (6:89:5)

B: acetonitrile/water/formic acid (80:15:5)

0.2 mL/min

280 and 320 nm

External standard (RT)

External standard calibration

Phenolic acid Wiczkowski et al. (2016)
HPLC–UV C18 RP (5 μm, 250 × 4.6 mm); 35 °C

A: acetic acid/water (2:98)

B: acetic acid/ acetonitrile/water (2:45:53)

1 mL/min; 10 μL

355 nm

External standard (RT)

External standard calibration

Flavonoid Yang et al. (2016)
HPLC–UV C18 RP (5 μm, 250 × 4.6 mm); 30 °C

A: acetic acid/water (0.15:99.85)

B: methanol

1 mL/min; 20 μL

280 nm

External standard (RT)

External standard calibration

Flavonoid and phenolic acid Park et al. (2017)
HPLC–MS C18 RP (5 μm, 250 × 4.6 mm); 30 °C

A: acetic acid/water (0.15:99.85)

B: methanol

1 mL/min; 20 μL

ESI negative mode

Scanning (100–1300 m/z)

External standard (RT) + MS spectra

External standard calibration

HPLC C18 RP (5 μm, 250 × 4.6 mm)

A: water

B: acetic acid/methanol (5:95)

1 mL/min; 5 μL

254 nm

External standard (RT)

External standard calibration

Flavonoid Jeong et al. (2018)
UPLC-MS C18 RP (1.7 μm, 50 × 2.1 mm); 40 °C

A: 0.1% formic acid in water

B: 0.1% formic acid in methanol

0.25 mL/min

ESI negative mode

External standard (RT) + MS spectra

External standard calibration

Internal standard (chrysin)

Flavonoid Koja et al. (2018)
HPLC–PDA C18 RP (5 μm, 250 × 4.6 mm); 40 °C

A: 0.1% formic acid in water

B: 0.1% formic acid in acetonitrile

0.8 mL/min; 20 μL

350 nm

External standard (RT)

External standard calibration

Flavonoid Nam et al. (2018a)
HPLC–PDA C18 RP (5 μm, 250 × 4.6 mm); 40 °C

A: 0.1% formic acid in water

B: 0.1% formic acid in acetonitrile

0.8 mL/min; 20 μL

350 nm

External standard (RT)

External standard calibration

Flavonoid Nam et al. (2018b)
HPLC–PDA C18 RP (5 μm, 250 × 4.6 mm); 40 °C

A: 0.1% formic acid in water

B: 0.1% formic acid in acetonitrile

1 mL/min; 5 μL

360 nm

External standard (RT)

External standard calibration

Flavonoid Jang et al. (2019)
HPLC–PDA C18 RP (5 μm, 250 × 4.6 mm); 40 °C

A: 0.1% formic acid in water

B: 0.1% formic acid in acetonitrile

0.8 mL/min; 10 μL

350 nm

External standard (RT)

External standard calibration

Flavonoid Mansur et al. (2019)
HPLC–Q–TOF/MS C18 RP (1.8 μm, 150 × 2.1 mm); 40 °C

A: 0.1% formic acid in water

B: 0.1% formic acid in acetonitrile

0.3 mL/min; 2 μL

ESI negative mode

Scanning (100–1000 m/z)

External standard (RT) + MS spectra

HPLC–UV C18 RP (5 μm, 250 × 4.6 mm); 30 °C

A: acetic acid/water (0.15:99.85)

B: methanol

1 mL/min; 20 μL

280 nm

External standard (RT)

External standard calibration

Flavonoid and phenolic acid Park et al. (2019)
UPLC-MS C18 RP (1.7 μm, 150 × 2.1 mm); 40 °C

A: 0.1% formic acid in water

B: 0.1% formic acid in acetonitrile

0.3 mL/min; 1 μL

ESI positive mode

External standard (RT) + MS spectra

External standard calibration

Flavonoid Chen et al. (2020)
HPLC–PDA C18 RP (5 μm, 250 × 4.6 mm); 40 °C

A: 0.1% formic acid in water

B: 0.1% formic acid in acetonitrile

0.8 mL/min; 10 μL

350 nm

External standard (RT)

External standard calibration

Flavonoid Mansur et al. (2020)
HPLC–UV C18 RP (5 μm, 250 × 4.6 mm)

A: water

B: 5% acetic acid in methanol

1 mL/min; 20 μL

254 nm

External standard (RT)

External standard calibration

Flavonoid Sim et al. (2020)
HPLC–PDA C18 RP (5 μm, 250 × 4.6 mm); 25 °C

A: 1% formic acid in water

B: 1% formic acid in acetonitrile

1 mL/min

370 nm

External standard (RT)

External standard calibration

Flavonoid Witkowicz et al. (2020)
HPLC–DAD Silica NP (7 μm, 150 × 3 mm)

A: water-formic acid (90:10)

B: acetonitrile/water/formic acid (85:10:5)

0.8 mL/min; 20 μL

External standard (RT)

External standard calibration

Internal standard (3,5-dichloro-4-hydroxybenzoic)

Flavonoid and phenolic acid Almuhayawi et al. (2021)
HPLC–MS/MS C18 RP (2.7 μm, 50 × 0.5 mm); 45 °C

A: water/formic acid (99.05:0.95)

B: acetonitrile/formic acid (99.05:0.95)

0.015 mL/min

ESI negative mode (MRM)

External standard (RT) + MS spectra

External standard calibration

Flavonoid and phenolic acid Dębski et al. (2021)
HPLC–UV C18 RP (5 μm, 150 × 4.6 mm); 35 °C

A: 1% phosphoric acid in water

B: methanol

1 mL/min

280 nm

External standard (RT)

External standard calibration

Flavonoid Hung et al. (2021)
HPLC–PDA C18 RP (5 μm, 250 × 4.6 mm); 40 °C

A: 0.1% formic acid in water

B: 0.1% formic acid in acetonitrile

1 mL/min; 5 µL

360 nm

External standard (RT)

External standard calibration

Flavonoid Jang et al. (2021)
HPLC–MS C18 RP (5 μm, 250 × 4.6 mm); 40 °C

A: 0.1% formic acid in water

B: 0.1% formic acid in acetonitrile

1 mL/min; 5 µL

ESI negative and positive mode

Scanning (200–1200 m/z)

External standard (RT) + MS spectra

HPLC–DAD C18 RP (3 μm, 150 × 2 mm); 25 °C

A: 5% acetonitrile + 0.1% o-phosphoric acid

B: 80% acetonitrile + 0.1% o-phosphoric acid

0.25 mL/min

350 nm

External standard (RT)

External standard calibration

Flavonoid Kalinová et al. (2021)
HPLC–MS C18 RP (2.7 μm, 150 × 2.1 mm); 30 °C

A: 0.1% aqueous formic acid

B: acetonitrile

0.25 mL/min; 20 µL

ESI negative mode

External standard (RT) + MS spectra

External standard calibration

Flavonoid and phenolic acid Živković et al. (2021)

aUFLC ultra-fast liquid chromatography, CE capillary electrophoresis, UPLC ultra-high performance liquid chromatography, DAD diode array detector, ECD electrochemical array detector, PDA photodiode array detector, Q-TOF/MS quadrupole time-of-flight/MS

bRP reversed-phase, NP normal phase

cSDS: sodium dodecyl sulfate; C-glycosyl flavones: orientin, isoorientin, vitexin and isovitexin

dRT: retention time comparison; ESI: electron spray ionization; MS spectra: data from MS analysis

eThe specific name for polyphenol analyte is described in Table 2