Table 3.

Extraction and detection methods for berberine in different herbal and biological matrixes.

Sample (weight)	Extraction method	Detection method	References
Dried stem powder Coscinium fenestratum (Gaertn.) (1 g)	Extraction solvents (ES): water, methanol–water (1:1. v/v), and methanol Sonication (15 min, room temperature) Centrifugation (2,800 rpm, 15 min) Filtration and evaporation Extracts resolubilization (methanol:water, 9:1 v/v)	HPLC - DAD Column: ODS, Chromolith, RP-18e,100 × 4.6 mm Mobile Phases: Methanol/Deionized Water (90:10, v/v) Flow: 0.5 mL/min, Temperature: 25°C UV Spectrophotometric Analysis	Akowuah et al., 2014
C. fenestratum (Gaertn.) (10 g)	ES: methanol Hot extraction: sample refluxed with ES for 3 h Filtration and evaporation. Extracts resolubilization (methanol)	TLC Adsorbent: Silica Gel GF 254 Solvent system: n-Butanol: Ethyl acetate: Acetic acid (2.5:1.5:1, v/v/v) Detection: 254 and 366 nm	Arawwawala and Wickramaar, 2012
	Cold extraction: sample extraction with ES for 24 h Filtration and evaporation. Extracts resolubilization (methanol)
Dried C. fenestratum (0.1 g)	ES: absolute methanol Cold extraction: sample extraction at −20°C Hot extraction: water bath sample extraction at 50°C ES: absolute ethanol Cold extraction sample extraction at −20°C Hot extraction: water bath sample extraction at 50°C Samples centrifugation (10 min at 10°C after cooling down) Samples filtration	HPLC Column: C18, 250 × 4.6 mm, 5 μm Mobile Phases: Acetonitrile/0.1% Trifluro-acetic acid (50:50, v/v) Detection: 344 nm Flow: 0.8 mL/min	Babu et al., 2012
C. fenestratum (1,000g) Capsules (containing 62.5 mg C. fenestratum)	ES: petroleum ether, chloroform, methanol (1L each) Soxhlet extraction: with each ES for 3 days at (30–40°C) ES: methanol (10 mL) Extraction for 1 h Filtration and evaporation Resolubilisation in methanol (5 mL)	HPLC Column: Luna C18, 150 × 4.6 mm, 5 μm, Phenomenex Mobile Phases: (A) Potassium dihydrogen phosphate (pH −2.5) and (B) Acetonitrile Detection: 220 nm Flow: 1 mL/min HPTLC Adsorbent: Silica Gel 60F 254 Solvent system: n-Butanol: Glacial acetic acid: Water (8:1:1, v/v/v) Detection: 350 nm for all measurements	Jayaprakasam and Ravi, 2014
Tinospora cordifolia (Willd.), Tribulus terrestris (L.), Emblica officinalis (Gaertn.) (3 g)	ES: chloroform Dried sample trituration with ammonia solution Drying at room temperature Extraction with ES for 1h Chloroform phase extraction with 5% sulfuric acid (x 3) Basification of acid extract with sodium carbonate (pH −9) Extraction of basified solution with chloroform (X 3) Evaporation of chloroform phase (temperature under 50°C) Residue solubilization with methanol	UV-VIS UV absorbance: 348 nm	Joshi and Kanaki, 2013
Cortex phellodendri (2 g)	Ultrahigh pressure extraction (UPE) Optimal parameters: ES: ethanol (69.1%), liquid-solid ratio−31.3, extracting pressure−243.30 MPa, extraction time−2 min	HPLC Column: Hypersil ODS C18, 250 × 4.6 mm, 5 μm Mobile Phases: (A) 0.3% triethanolamine aqueous solution (pH − 3.5) Detection: 265 nm Temperature: 30°C Flow: 1 mL/min	Guoping et al., 2012
Rhizome of Coptis chinensis Franch (1 g)	Supercritical fluid extraction Extraction time: up to 3 h Temperature: 60°C Pressure: from 200 to 500 bar Flow-rate of carbon dioxide (gaseous state): 1 L/min Flow-rate of modifier: 0.4 mL/min. Organic solvent modifier systems: ethanol-modified supercritical carbon dioxide, methanol-modified supercritical carbon dioxide, 1,2-propanediol-modified supercritical carbon dioxide, 5% Tween 80 in methanol-modified supercritical carbon dioxide, 5% Tween in ethanol-modified supercritical carbon dioxide	HPLC Column: Diamonsil C18, 250 × 4.6 mm, 5 μm Mobile Phases: 33 mM Potassium dihydrogen phosphate : acetonitrile (70:30, v/v) Detection: 345 nm Flow: 1 mL/min	Liu et al., 2006
	Soxhlet extraction ES: hydrochloric acid: methanol (1: 100, v/v) Time: 8 h
Cortex pellodendri amurensis (1 g)	Ultrahigh pressure extraction ES: ethanol (50 %), liquid-solid ratio −30: 1, extracting pressure −400 MPa, extraction time −4 min, extraction temperaturte −40°C Ultrasonic extraction ES: 70% ethanol Sample soaking for 24 h in 40 ml ES Sonic extraction for 60 min at 30°C	HPLC- DAD Column: Daisopak SP-120-5-ODS_BP, 250 × 4.6 mm, 5 μm Mobile Phases: (A) acetonitrile and (B) phosphoric acid: water (0.7:100, v/v) Detection: 345 nm Temperature: 25°C Flow: 1 mL/min	Liu et al., 2013
	Heat reflux extraction ES: 70% ethanol Sample soaking for 24 h in 40 ml ES Sample extraction for 4 h at boiling state
	Soxhlet extraction ES: 70% ethanol Sample soaking for 24 h in 40 ml ES Sample extraction: 4 h
Goldenseal (Hydrastis canadensis L.) (2, 5, 5 g)	Pressurized hot water extraction ES: water at 140°C, Optimal parameters: pressure: 50 bars and flow rate: 1 mL/min, Time: 15 min Reflux extraction ES: methanol (200 mL) Sonication: 4 h at 80°C Ultrasonic extraction ES: methanol (50 mL) Reflux: 6 h with continuous stirring	HPLC-DAD Column: Zorbax eclipse Plus C 18, 75 x 4.6 mm, 3.5 μm Mobile phases: (A) 0.1 % Formic Acid (pH 2.7) and (B) methanol Detection: 242 nm Temperature: 35°C Flow: 1 mL/min MS Detection: ESI (+) Capillary temperature: 200°C, Sheath gas: 80, Capillary voltage: 20 V, Tube lens voltage: 5V	Mokgadi et al., 2013
Berberis aristata DC (1.5 g), Berberis aristata herb extract (0.1 g), Ayurvedic form (6 g)	Crude herb reflux extraction ES: methanol (100 mL) for 1 h in a water bath Filtratio Reextraction with ES (50 mL) for 30 min (× 2) Filtrates combination and concentration to 50 mL Herb extracts ultrasonic extraction ES: methanol (up to 10 mL) Sonication Filtration	HPLC Column: Zorbax ODS II, 250 x 4.6 mm, 5 μm Mobile phase: potassium hydrogen phosphate buffer (pH 2.5)/ acetonitrile Detection: 346 nm Temperature: 40°C Flow: 1 mL/min	Singh R. et al., 2010
	Ayrvedic form ultrasonic extraction ES: methanol (up to 25 mL) Sonication
Berberis aristata DC root	Soxhlet extraction ES: ethanol Berberine isolation Ethanolic extract concentration to obtain a syrup mass Dissolvation in hot water and filtration Acidification (36.5% w/v hydrochloric acid) Cool: ice bath - 30 min, overnight in refrigerator	HPTLC Stationary phase: precoated silica gel 60GF254 Mobile phases: n-butanol: glacial acetic acid: water (12:3:4 v/v/v) Temperature: 33 ± 5°C Detection: 350 nm	Patel, 2013
Mahonia manipurensis (Takeda) stem bark (100 g)	Cold extraction ES: 80% methanol (1,000 mL) Stirring at room temperature Extract concentration	TLC Stationary phase: precoated silica gel G F254 Mobile phase: hexane: ethyl acetate: methanol (56:20:5) Fraction purification: positive test using Dragendroff's reagent Further analysis of purified fraction Mobile phase: chloroform: ethyl acetate: diethylamine: methanol: 20% ammonium hidroxide (6:24:1.5:6:0.3)	Pfoze et al., 2014
		HPLC Column: Water Symmetry C18, 250 x 4.6 mm, 5 μm Mobile phase: methanol/ formic acid buffer (0.1%, v/v) Detection: 346 nm Flow: 1 mL/min
		UV-VIS UV spectra: 200–500 nm
		ESI-MS
Coscinium fenestratum (100 g)	Maceration ES: 80% ethanol (500 ml), 160 h Shaken: 80 h (200 rpm), stand: 80 h Reextraction: 48 h, shaken: 24 h, stand: 24 h Combined extracts concentration Evaporation to dryness (dry extract) Resolubilisation in 80% ethanol (10 mg dry extract/mL)	TLC Stationary phase: Silica gel GF254 Mobile phase: ethyl acetate : butanol : formic acid : water (50:30:12:10); Detection: 366 nm	Rojsanga and Gritsanapan, 2005
Argemone mexicana	Soxhlet extraction ES: methanol Evaporation to dryness Resolubilisation in methanol (known concentration)	HPTLC Stationary phase: precoated silica gel 60F254 Mobile phases: toluene: ethyl acetate (9:3, v/v). Detection: 266 nm	Samal, 2013
Tinospora cordifolia (20 g)	Microwave assisted extraction (MAE) ES: 80% ethanol Irradiation power: 60%, Extraction time: 3 min Soxhlet extraction ES: ethanol, for 3 h Filtration Concentration	HPTLC Mobile phases: methanol: acetic acid: water (8: 1: 1, v/v/v). Detection: 366 nm	Satija et al., 2015
	Maceration ES: ethanol (200 mL), 7 days, occasional stirring
Berberis aristata, Berberis tinctoria (800 g)	Hot extraction ES: methanol (2.5 L) (X2) Extraction time: 3 h Temperature: 50°C Extract concentration under vacuum	HPLC Column: Unisphere C18, 150 x 4.6 mm, 5 μm Mobile phase: (A) 0.1% trifluoroacetic acid and (B) acetonitrile (60:40, v/v) Detection: 350 nm Temperature: 30°C Flow: 1 mL/min	Shigwan et al., 2013
Coptis chinensis Franch. (1g)	Acid assisted extraction ES: several inorganic acids (hydrochloric acid, phosphoric acid, nitric acid, and sulfuric acid) and one organic acid (acetic acid) Extraction time:1–8 h, Acid concentrations: 0–1% Solvent to sample ratios: 20–60 mL/g Maceration at 25°C Filtration Dilution to 100 mL final volume	HPLC Column: XTerra C18, 250 x 4.6 mm Mobile phase: (A) acetonitrile and (B) 25 mmol/L potassium dihydrogen phosphate,(27:75, v/v) Detection: 345 nm Temperature: 30°C	Teng and Choi, 2013
	Soxhlet extraction ES: 50% ethanol (100 mL), 4 h at 70°C Extract evaporation to dryness Resolubilization in ES (up to 100 mL final volume)
	Heating reflux extraction ES: 50% ethanol Soaked for 1 h Extraction: 4 h at 70°C (heated water bath) Filtration Dilution (up to 100 mL final volume)
Rabbit plasma (100 μl)	Mixing 100 μl sample with 3% formic acid in acetonitrile (200 μl) Vortex: 30 s Centrifugation: 10 min at 4°C Evaporation of supernatant: under nitrogen stream at 40°C	LC-ESI-MS HPLC system Column: Capcell Pakc18 MG, 100 × 2.1 mm, 5 μm with Security Guard C18, 4 × 2 mm, 5 μm Mobile Phases: (A) 0.4% formic acid solution and (B) 0.2 % formic acid solution in methanol (60:40, v/v)	Liu et al., 2011
	Residue solubilization: in 100 μl of 20% methanol	Temperature: 25°C Flow: 0.4 mL/min MS detection: Source: ESI (+) Quantification: MRM mode
Rat plasma	Solid phase extraction (SPE) Cartridges: Oasis HLB (1 cc, 30 mg) Pre-conditioning: 2 mL methanol Equilibrtating:	UPLC-MS/MS UPLC system Column: 120 EC-C18, 50 × 4.6 mm, 2.7 μm with Security Guard C18, 4 × 2 mm, 5 μm Mobile Phases: (A) 10 mM ammonium acetate in water (pH- 4.5) and (B) acetonitrile Temperature: 35°C Flow: 0.8 mL/min MS detection: Source: ESI (+) Quantification: MRM mode	Liu M. et al., 2015
Rat plasma Rat tissue	Rat plasma ES: methanol Mixing sample (200 μl) with internal standard (40 μl) and ES (560 μl) Vortex: 20 s Centrifugation: 10 min, 12,000x g Filtration	UPLC-MS/MS UPLC system Column: Acquity BEH C18, 50 × 2.1 mm, 1.7 μm Mobile Phases: (A) acetonitrile and (B) formic acid: water (0.1:99.9, v/v) Flow: 0.25 mL/min MS detection: Source: ESI (+) Quantification: MRM mode	Wang et al., 2016
	Rat tissue Grinding: 3 mL physiological saline with 600 mg tissue Centrifugation: 10 min, 12,000x g, 4°C Mixing supernatant (200 μl) with internal standard (40 μl) and ES (560 μl) Vortex: 20 s Centrifugation: 10 min, 12,000x g Filtration
Rat plasma	Evaporation of 10 ul IS in the working tube Mixing sample (200 μl) with internal evaporated standard Vortex: 1 min Mixing sample with 10 μl 1% formic acid and 200 μl acetone Vortex: 2 min Centrifugation: 10 min, 10,000 rpm Mixing supernatant with 200 μl methanol Vortexing, centrifugation Mixing supernatant wit 400 μl acetonitrile Vortexing, centrifugation Evaporation to dryness (37°C, under nitrogen stream) Resolubilization in methanol	LC-MS/MS LC system Column: Zorbax Eclipse XDB-C18, 150 × 2.1 mm, 3.5 μm Mobile Phases: (A) acetonitrile and (B) water with 1% acetic acid and 0.001 mol/L ammonium acetate Flow: 0.2 mL/min MS detection: Source: ESI (+) Quantification: MRM mode	Xu et al., 2015
Rat plasma	ES: 90% methanol Mixing sample (100 μl) with internal standard (10 μl) and ES (100 μl) Vortex: 1 min Centrifugation: 10 min, 12,000 rpm, 4°C Supernatant evaporation to dryness under nitrogen stream Resolubilization (100 μl ES)	UPLC-MS/MS UPLC system Column: Acquity UPLC BEH C18, 50 × 2.1 mm, 1.7 μm Mobile Phases: (A) formic acid: water (0.1:99.9, v/v) and (B) acetonitrile Flow: 0.4 mL/min MS detection: Source: ESI Quantification: MRM mode	Yang et al., 2017