Liquid Chromatography

Liquid chromatography is currently one of the important methods for analysis of trace amount of substance component. The two-phase in the chromatographic analysis refers to immobile phase having a large specific surface area and mobile phase carrying mixture to be separated to flow through the immobile phase. Chromatography using liquid as mobile phase is called as liquid chromatography.

Liquid chromatography is one kind of chromatography method with the adopted mobile phase being liquid. Owing to the liquid contained in the column gets higher resistance than gas in the column, in order to increase the speed of analysis, people usually adopt high-pressure fluid way, so called "high-pressure liquid chromatography." According to the difference of the mobile phase, it can be divided into liquid - solid chromatography and liquid - liquid chromatography; according to the difference type of equilibrium of the immobile phase and the mobile phase, it can be divided into adsorption chromatography, partition chromatography, ion exchange chromatography and gel chromatography. It can be used for the separation and analysis of organic, inorganic and polymer liquid.

Brief History
In 1906, the Russian botanist Tswett has for the first time applied column chromatography for the separation plant pigments, and is one of oldest liquid chromatography. In the recent 30 years, people have developed various kinds of chromatography and become an important part of the chemical analysis.
① classical liquid chromatography: mobile phase flow downward from the upper end of the column tube through its own gravity; collect the effluent one tube by tube from the lower portion of the column at the outlet and then apply other methods of measurement with slow analysis rate and low efficiency;
② Thin-layer chromatography : it appeared in around 1940s, easy to operate and have obtained improvement on the speed of analysis, but has poor reproducibility and difficult for quantitative analysis;
③ High performance liquid chromatography: after the early 1960s, J. C. Gidelings had amended the gas chromatography theory to liquid chromatography and had laid the foundation for its modernization. Thereafter, technically people had used high-pressure pump, high efficiency immobile phase and a high sensitivity detector and had developed the high performance liquid chromatography with high speed analysis, high separation efficiency and automatic operation. In 1971, R. A. Henry has first applied it for pesticide analysis.

High performance liquid chromatography
High performance liquid chromatography is one of the most important separation method of modern analytical chemistry. It is originated in the classical liquid chromatography with the basic approach as below: use single solvent of certain polarity or mixed solution of different proportions as mobile phase; pump the mobile phase into the column equipped with a filler; after the injected tested sample has been brought into the column by the mobile phase, each components then entered the detector one after another; use recorder or data processing device to  record the chromatogram or perform data processing to obtain the measurement results. Owing to the application of the filler particles of various characteristics and pressurized liquid mobile phase, the separation method has high performance and rapid analysis features.

High performance liquid chromatography analysis is suitable for the determination and measurement of the drugs which can be separated on column equipped with certain filler, in particular for the determination of multi-component drugs, impurities inspection and the determination of macromolecules. Some drugs need to be subject to derivatization reaction prior to or after the chromatographic separation in order to be isolated or detected. The commonly used filler of the column include: silica gel used for normal phase chromatography; chemically bonded immobile phase, depending on the bonding groups, can be used in either reverse phase or normal phase chromatography; ion exchange filler is used for ion exchange chromatography; macroporous filler with certain aperture is used for exclusion chromatography.

High performance liquid chromatography consists of the basic pump, injector, column, detector and the processing system of the chromatographic data. The most commonly used detector is for detecting UV and visible light; other detectors also include refractive index detector and evaporative light scattering detector and so on. The collection and processing of the chromatography information often proceed in information workstation and integrators. Gradient elution can be obtained through program control with two pumps or single pump plus proportional valve.

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Structure Chemical Name CAS MF
FERROCENEACETIC ACID FERROCENEACETIC ACID 1287-16-7 C12H12FeO2
4-Chloro-7-nitrobenzofurazan 4-Chloro-7-nitrobenzofurazan 10199-89-0 C6H2ClN3O3
Water Water 7732-18-5 H2O
2-Methylbutane 2-Methylbutane 78-78-4 C5H12
Fluorescamine Fluorescamine 38183-12-9 C17H10O4
DME DME 110-71-4 C4H10O2
PETROLEUM ETHER PETROLEUM ETHER 8032-32-4 unspecified
NAM NAM 49759-20-8 C17H10N2O2
ABD-F ABD-F 91366-65-3 C6H4FN3O3S
4-(N,N-DIMETHYLAMINOSULFONYL)-7-FLUORO-2,1,3-BENZOXADIAZOLE 4-(N,N-DIMETHYLAMINOSULFONYL)-7-FLUORO-2,1,3-BENZOXADIAZOLE 98358-90-8 C8H8FN3O3S
4-Nitro-7-(1-piperazinyl)-2,1,3-benzoxadiazole 4-Nitro-7-(1-piperazinyl)-2,1,3-benzoxadiazole 139332-66-4 C10H11N5O3
2-METHOXY-2,4-DIPHENYL-3(2H)-FURANONE 2-METHOXY-2,4-DIPHENYL-3(2H)-FURANONE 50632-57-0 C17H14O3
O-(4-NITROBENZYL)-N,N'-DIISOPROPYLISOUREA O-(4-NITROBENZYL)-N,N'-DIISOPROPYLISOUREA 2978-11-2 C14H21N3O3
(S)-(+)-4-NITRO-7-(3-AMINOPYRROLIDIN-1-YL)-2,1,3-BENZOXADIAZOLE (S)-(+)-4-NITRO-7-(3-AMINOPYRROLIDIN-1-YL)-2,1,3-BENZOXADIAZOLE 143112-52-1 C10H11N5O3
N-SUCCINIMIDYL 4-NITROPHENYLACETATE N-SUCCINIMIDYL 4-NITROPHENYLACETATE 68123-33-1 C12H10N2O6
Fluorescein 6-isothiocyanate Fluorescein 6-isothiocyanate 18861-78-4 C21H11NO5S
9-(BROMOMETHYL)ACRIDINE 9-(BROMOMETHYL)ACRIDINE 1556-34-9 C14H10BrN
2,3-DIHYDRO-6-ISOTHIOCYANATO-1,4-PHTHALAZINEDIONE 2,3-DIHYDRO-6-ISOTHIOCYANATO-1,4-PHTHALAZINEDIONE 107807-39-6 C9H5N3O2S
DABSYL HYDRAZINE DABSYL HYDRAZINE 72565-41-4 C14H17N5O2S
(R)-(-)-NBD-APY (R)-(-)-NBD-APY 143112-51-0 C10H11N5O3
N-CHLOROMETHYL-4-NITROPHTHALIMIDE N-CHLOROMETHYL-4-NITROPHTHALIMIDE 54455-34-4 C9H5ClN2O4
DBD-ED DBD-ED 189373-41-9 C10H15N5O3S
1-(4-NITROBENZYL)-3-P-TOLYLTRIAZENE 1-(4-NITROBENZYL)-3-P-TOLYLTRIAZENE 60259-80-5 C14H14N4O2
(1R,2R)-2-(NAPHTHALENE-2,3-DICARBOXIMIDO)CYCLOHEXANECARBOXYLIC ACID (1R,2R)-2-(NAPHTHALENE-2,3-DICARBOXIMIDO)CYCLOHEXANECARBOXYLIC ACID 642995-15-1 C19H17NO4
1-BENZYL-3-P-TOLYLTRIAZENE 1-BENZYL-3-P-TOLYLTRIAZENE 17683-09-9 C14H15N3
108-32-7 108-32-7 108-32-7
(1R,2R)-2-(ANTHRACENE-2,3-DICARBOXIMIDO)CYCLOHEXANECARBOXYLIC ACID (1R,2R)-2-(ANTHRACENE-2,3-DICARBOXIMIDO)CYCLOHEXANECARBOXYLIC ACID 446044-44-6 C23H19NO4
(1S,2S)-2-(NAPHTHALENE-2,3-DICARBOXIMIDO)CYCLOHEXANECARBOXYLIC ACID (1S,2S)-2-(NAPHTHALENE-2,3-DICARBOXIMIDO)CYCLOHEXANECARBOXYLIC ACID 642995-16-2 C19H17NO4
4-(N,N-DIMETHYLAMINOSULFONYL)-7-[5-(SUCCINIMIDYLOXYCARBONYL)PENTYLAMINO]-2,1,3-BENZOXADIAZOLE 4-(N,N-DIMETHYLAMINOSULFONYL)-7-[5-(SUCCINIMIDYLOXYCARBONYL)PENTYLAMINO]-2,1,3-BENZOXADIAZOLE 1858255-08-9 C18H23N5O7S
3-BROMOMETHYL-7-METHOXY-1,4-BENZOXAZIN-2-ONE 3-BROMOMETHYL-7-METHOXY-1,4-BENZOXAZIN-2-ONE 124522-09-4 C10H8BrNO3
(S)-(-)-NBD-PRO-COCL (S)-(-)-NBD-PRO-COCL C11H9ClN4O4
5-Aminofluorescein 5-Aminofluorescein 3326-34-9 C20H13NO5
DBD-PZ DBD-PZ 139332-64-2 C12H17N5O3S
(1S,2S)-2-(ANTHRACENE-2,3-DICARBOXIMIDO)CYCLOHEXANECARBOXYLIC ACID (1S,2S)-2-(ANTHRACENE-2,3-DICARBOXIMIDO)CYCLOHEXANECARBOXYLIC ACID 446044-45-7 C23H19NO4
DBD-COCL DBD-COCL 156153-43-4 C11H13ClN4O4S
4-(5-CARBOXYPENTYLAMINO)-7-(N,N-DIMETHYLAMINOSULFONYL)-2,1,3-BENZOXADIAZOLE 4-(5-CARBOXYPENTYLAMINO)-7-(N,N-DIMETHYLAMINOSULFONYL)-2,1,3-BENZOXADIAZOLE 1820741-40-9 C14H20N4O5S
DIPENTYLAMINE ACETATE SOLUTION DIPENTYLAMINE ACETATE SOLUTION 211676-91-4 C12H27NO2
NALPHA-(5-FLUORO-2,4-DINITROPHENYL)-L-LEUCINAMIDE NALPHA-(5-FLUORO-2,4-DINITROPHENYL)-L-LEUCINAMIDE 178065-29-7 C12H15FN4O5
DMEM DMEM
Succinimidyl (2R)-6-(Tetrahydro-2H-pyran-2-yloxy)-2,5,7,8-tetramethylchroman-2-carboxylate Succinimidyl (2R)-6-(Tetrahydro-2H-pyran-2-yloxy)-2,5,7,8-tetramethylchroman-2-carboxylate 1069137-73-0 C23H29NO7
DBD-NCS DBD-NCS 147611-81-2 C9H8N4O3S2
DIPROPYLAMINE ACETATE SOLUTION DIPROPYLAMINE ACETATE SOLUTION 114389-69-4 C8H19NO2
AABD-SH AABD-SH 254973-02-9 C8H7N3O2S
NALPHA-(5-FLUORO-2,4-DINITROPHENYL)-D-LEUCINAMIDE NALPHA-(5-FLUORO-2,4-DINITROPHENYL)-D-LEUCINAMIDE 178065-30-0 C12H15FN4O5
NBD-CO-HZ NBD-CO-HZ 221263-97-4 C9H10N6O4
NBD-COCL NBD-COCL 140164-85-8 C9H7ClN4O4
(HYDRAZINOCARBONYL)FERROCENE (HYDRAZINOCARBONYL)FERROCENE 12153-28-5 C11H12FeN2O10*
NBD-H NBD-H 131467-87-3 C6H9N7O3
DBD-CO-HZ DBD-CO-HZ 179951-63-4 C11H16N6O4S
DBD-H DBD-H 131467-86-2 C8H11N5O3S
(R)-(+)-NBD-PRO-COCL (R)-(+)-NBD-PRO-COCL C11H9ClN4O4
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