Table 6.

Summary of different methods and representative sample preparation procedures for analysis of markers in Pinellia ternata (Thunb.) Breit.

Method	Analytes	Sample preparation	Column	Mobile phase	Reference
UV	Beta-Sitosterol	0.4 g of PR powder was mixed with 20 mL ethyl acetate, and cold soak overnight, take the filtrate. The filtrate was extracted twice, the ethyl acetate was recovered, and the residue was diluted to 10 mL with ethyl acetate.			[148]
TLC	Pinellia ternata Pinellia pedatisecta	1 g PR power was mixed with 20 mL of ethanol and then subjected to refluxed extraction for 1 h and evaporated to dryness. The residue was dissolved in 1 mL of ethanol.	Silicone G Thin Layer Plates	Trichloromethane-methanol-water (6:4:0.5)	[149]
ELISA	Pinellia ternata lectin	1 g PR power was mixed with 20 mL of ultrapure water and shaking for 2 h, and centrifuged at 8000 r/min for 15 min. The residue was re-extracted twice under the same conditions and supernatants were combined.			[150]
ELISA	PTL	1 g PR power was mixed with 20 mL of PBS and shaking for 1 h, and centrifuged at 8000 r/min for 15 min. The residue was re-extracted twice under the same conditions and supernatants were combined.			[151]
UPLC-MS/MS	Lysophosphatidylcholines Sanedrine Trigonelline Choline Protocatechuic acid Protamine sulfates Homogentisic acid Chrysophanic acid	5 g of PR、PRPCA、PRPZA and PRP power ware mixed with 50 mL of methanol and 10 L of internal standard solution, and reaction was extracted twice of 30 min each time, centrifuged at 10000 r/min for 15 min, and then the supernatants were combined.	Waters Acquity UPLC HSST3 (100 mm × 2.1 mm, 1.8 μm)	methanol (A)-water (B) with gradient elution	[152]
UPLC-Q-TOF-MS/MS	Alkaloids Flavonoids Amino acids Others	2 g PR powder was immersed in 70 % ethanol (20 mL) for 0.5 h, then extracted under reflux for 1 h, filtered, extracted with 70 % methanol (10 mL) for 0.5 h, combined filtrate.	shim-pack xR-ODSⅢ column (2.1 mm × 75 mm, 1.6 μm)	acetonitrile (A)-0.1 % formic acid in water (B) with gradient elution	[153]
HPLC	Uracil 6-Hydroxypurine Uridine Guanosine Thymidine	1 g of PR powder was mixed were 50 mL of water, weigh the mass, sonicate for 30 min, filter and take the filtrate.	Agilent Zorbax SB C18 column (4.6 mm × 250 mm, 5 μm)	methanol (A)-water (B) with gradient elution	[154]
HPLC	Uracil Cytidine Uridine Inosine Guanosine Thymidine Adenosine	0.2 g of PR powder was mixed were 10 mL of water, ultrasonic extraction for 1 h, centrifuged at 3000 r/min for 10min, and the supernatant was taken.	Agilent Zorbax SB Cs (4.6 mm × 250 mm, 5 μm)	water (A)-methanol (B) with gradient elution	[155]
HPLC	Inosine Guanosine Adenosine Succinic acid Ephedrine Hydrochloride	2 g of PR powder was mixed with 20 mL of ultrapure water, ultrasonic extraction for 45 min, 10000 r/min−1 centrifugation for 5 min, take the supernatant 10 mL, recover the solvent to dry, add the appropriate amount of ultrapure water to make the dissolution, transferred to a 10 mL measuring flask, with the use of purified water to the gradient of the volume, shaking well, and the extract was then extracted.	Agilent Eclipse XDB C18 column (4.6 mm × 250 mm, 5 μm)	acetonitrile (A)- water (B) with gradient elution	[156]
HPLC	Inosine Guanosine Thymidine Adenosine Trigonelline	1 g of PR powder was mixed with 50 mL ultrapure water, ultrasonic extraction was performed 3 times, each time for 45 min. 12000 r/min centrifugation was performed for 10 min, and the supernatants of the 3 extractions were combined.	C18 column (150 mm × 4.6 mm, 5 μm)	methanol and water (3:97)	[157]
UPLC	Calcium oxalate	Precisely weigh the appropriate amount of PR powder, add ultra-pure water vortex mixing, set to 60 °C aqueous solution, set to 60 °C water bath heating and stirring, centrifuged at 8000 r/min for 5 min, aspirate the supernatant, the same method for 2 times: combined supernatant and transferred to a 10 mL measuring flask, add water to set volume.	Acquity UPLC BEH Cl8 column (50 mm × 2.1 mm, 1.7 μm)	0.5 % (NH4) H2PO4	[158]
RP-HPLC	Oxalic acid Citric acid Malic acid Succinic acid	2 g of PR powder was mixed with 40 mL ultrapure water, ultrasonic extraction 2 times, each time 2 h, filtration, combined filtrate, with ultrapure water to 100 mL, precision measurement of 20 mL, add concentrated ammonia to adjust pH 11.5, with 3 times the amount of extract of ethyl acetate (60 mL) extraction for 3 times, collection of alkaline solution, acidified with phosphoric acid to pH 2.0, and then 3 times the amount of ethyl acetate extraction for 5 times. Then the extract was extracted with 3 times amount of ethyl acetate for 5 times, collected the ethyl acetate, dried by rotary evaporation, dissolved in ultrapure water and concentrated to 25 mL.	Gemini-C18 column (4.6 mm × 250 mm, 5 μm)	methanol and 0.03 % ammonium dihydrogen phosphate buffer (97:3)	[159]
Potentiometric titration	Total acid	5 g PR power was mixed with 95 % ethanol and then subjected to refluxed extraction for 1 h, extracted three times, filtered and 0.1 mol/L NaOH aqueous solution (10 mL), and then subjected to ultrasonic extraction for 30 min.	NaOH, HCL titration solution		[160]
microchip electrophoresis with electrochemical detection	Guanosine Methionine Glycine 3,4-dihydroxybenzaldehyde Homogentisic acid	2 g PR power was mixed with 6 mL of the extraction solvent containing C2H5OH/DI H2O (1:1, v/v) and then subjected to ultrasonic extraction for 20 min.			[161]
HPLC	Uridine Adenine Guanosine Adenosine	1 g PR powder was mixed with 25 mL of water, sonicate for 30 min, cool to room temperature, shake well and centrifuge at 12000 r/min for 10 min, take the supernatant.	Shim-pack Scepter C18-120 column（250 mm × 4.6 mm, 5 μm）	10 % acetonitrile (A)-water (B) with gradient elution	[162]
GC-MS	Fat-soluble ingredients	500 g of PR powder, 3 times with 3 L methanol osmosis drip extraction, each 4 d, respectively, combined extracts, 38 concentrated under reduced pressure into a crude extract. The crude extract was dispersed into 600 mL of distilled water, and then extracted with 600 mL of petroleum acyl (60～90 °C) for 3 times, concentrated at 38 °C under reduced pressure, and then freeze-dried by freeze-dryer to obtain the extract. 1 mg of each extract was dissolved in 10 mL of methanol and sonicated.	VARIAN VF-5 ms column (30 m × 0.25 mm × 0.25 μm)	carrier gas: N2	[163]
MLPA	PR Pinellia pedatisecta Pinellia cordata	The surface of the herbs was scrubbed with 75 % ethanol and the surface cortex was scraped off, 30–50 mg of the dried samples were placed in 2 mL EP tubes and 2 small steel beads were added, and then the samples were ground into powder for 2 min in a high-throughput tissue grinder, and then the samples were extracted by using the Plant Genomic DNA Extraction Kit, and the concentration and purity of the DNA were determined by using the NanoDrop-20O0 Ultra-Micro Spectrophotometer.			[10]
PCR-RFLP	PR Arisaema erubescens	By comparing the rbcL sequences of Arisaema erubescens, PR and their mixed pseudo-products, the specific cleavage sites HaeIII and DraⅠ were selected for Arisaema erubescens and PR, respectively, and were identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)			[164]
ATR-FTIR	Pinellia ternata of different origins	The powder of pinellia ternata was finely ground in an agate mortar and pestle under dry conditions.			[165]