Multidimensional Chromatography and Its Applications: Comparison
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Please note this is a comparison between Version 2 by Catherine Yang and Version 1 by Mayeen Uddin Khandaker.

Food, drugs, dyes, extracts, and minerals are all made up of complex elements, and utilizing unidimensional chromatography to separate them is inefficient and insensitive. This has sparked the invention of several linked chromatography methods, each of them with distinct separation principles and affinity for the analyte of interest. Multidimensional chromatography consists of the combination of multiple chromatography techniques, with great benefits at the level of efficiency, peak capacity, precision, and accuracy of the analysis, while reducing the time required for the analysis.

  • chromatographic techniques
  • industrial application
  • efficiency
  • food

1. Foods, Flavors, and Fragrances

Natural complex mixtures are constituents of food, flavor, and fragrance products, with analysis of these matrices being carried out in [40][1] (Figure 1 and Table 1).
Figure 1.
Representative scheme with the most important applications of multidimensional chromatography.
  • Qualitative and/or quantitative determination of certain classes of constituents;
  • Quality and authenticity control of the product;
  • Adulteration or contamination detection.
One-dimensional chromatography does not provide sufficient selectivity for the separation of the complex, as there are problems of overlapping peaks, variation in selectivity, or even when the components are present in trace amounts, hampering the selectivity [41,42][2][3]. On the contrary, multidimensional chromatography provides an advanced pre-separation effect before analyte separation and quantification, ultimately overcoming the problems related to peak overlapping and trace element detection [43,44][4][5].

1.1. Gas Chromatography (GC–GC or MDGC)

A large number of food products, flavorings, and fragrances consist of chiral compounds that, on detection, present overlapping peaks and selectivity problems. The distribution of enantiomers is useful for the identification of adulterants, controlling the fermentation process and storage effects. GC–GC can be used for the enantiomeric determination of these products, but pre-selection is crucial due to overlapping peaks. The pre-chiral columns are being used before the main chiral column just to avoid overlapping peaks. This method is used to determine products’ authenticity [45][6].

1.2. High-Performance Liquid Chromatography (MD–HPLC)

Multidimensional HPLC has high separation power; hence it is used for the separation of complex molecules [18,46][7][8]. Heart-cut, on-column concentration, or trace enrichment are different types of techniques used to promote the application of multidimensional HPLC [47][9]. Using multidimensional HPLC, the researchers have been able to determine the levels of complex B vitamins in protein food, on-column vitamin concentrations in food matrixes, molasses sugars, malathion in tomato plants, lemonin in grapefruit peels, and other polar pesticides, with detection limits being as low as 0.1 g/L [48,49][10][11].

2. Biomedical and Pharmaceutical Applications

Multidimensional chromatography is used widely in the field of the biomedical and pharmaceutical industry due to the complex nature of the analyte and decreased amount present in the biological matrix [50,51,52][12][13][14] (Figure 1). High selectivity and accurate systems are needed with high reliance to determine such compounds [53,54][15][16].
LC is well established in these industries and very well adopted, so LC remains a component for separation and quantification purposes, along with other systems. Online and offline approaches are being used for coupling systems, where the online approach provides greater accuracy and precision along with shorter analysis time [55][17].
Charged, polar, thermolabile, non-volatile, and high-molecular-weight compounds are just some of the clinically relevant chemicals that can be directly examined by LC. Antibiotics, retinoids, methotrexate, codeine, psilocin, and almokalant have all been studied using this method, as well as anabolic steroids, morphine, and clozapine [56][18].

3. Industrial Chemicals and Polymer Applications

Industrial chemicals and associated materials have also been subjected to multidimensional chromatography analysis (Figure 1). Coal tar, antiknock additives in gasoline, light hydrocarbons, trihaloalkanes and trihaloalkenes in industrial solvents, soot, particle extracts, and different industrial compounds that may be found in gasoline and oil samples have been studied via multidimensional chromatography [57][19].
The rapid identification of polymer additives, such as antioxidants, lubricants, flame retardants, waxes, and UV stabilizers was made possible by a multidimensional system using capillary SEC–GC–MS [58][20]. Chemical additives, such as hydrocarbons and alcohols, for example, may not only be analyzed in the context of polymer chemistry but also in the context of food chemistry using the same methodologies. The study of polyaromatic hydrocarbons in food oils has also proven very important. Moreover, a polystyrene matrix has been employed for the investigation of polymer additives using SEC and GC [59][21].

4. MDC in Environmental Analysis

Environmental analysis relies heavily on multidimensional chromatography. The wide range of polarity of the constituents and a large number of isomers or congeners with similar or equal retention properties make it impossible to separate environmental samples using a single chromatographic approach (Table 1). Polycyclic aromatic hydrocarbons (PAHs), pesticides, and halogenated chemicals are all persistent organic pollutants that can be found in the air, water, soil, and sediment, and multidimensional chromatography has been used to analyze such contaminants [60][22]. Among them are polychlorinated dibenzo-dioxins, dibenzofurans, polychlorinated biphenyl, terphenyls, naphthalene, alkanes, organochlorine insecticides, and the brominated flame retardants, polybrominated biphenyls and polybrominated diphenyl ethers, which are some of the most representative [61][23].

5. Forensic Toxicological Applications

Analyses in forensic and toxicological sciences involve the identification of complexes that signal sickness, poisons, and a wide range of unlawful acts as examples [62][24] (Figure 1). Tissues, urine, plasma, serum, hair, arson debris, and fragments of other objects are often discovered to contain analytes [63,64][25][26]. For such purposes, LC–GC has been used for the analysis of numerous toxic complexes in biological samples, such as plasma and tissues, whereas GC–GC has been used for complex samples with exceptional selectivity. For example, organic pesticides and polychlorinated biphenyls (PCBs) may be separated and analyzed using online LC–GC, which can also be used for lipid matrices. In addition, the analysis of illicit growth hormones in corned beef can be performed using LC linked to two-dimensional GC [65][27]. Specifically, LC–LC has found its extensive use for the analysis of biological matrices, while GC–GC has special applications in determining drugs with specific selectivity. More recently, micellar HPLC has been used for the analysis of cardiovascular drugs from urine [66][28].
Multidimensional LC has also been used to determine ursodeoxycholic acid and its conjugates in serum, while multidimensional GC is used to separate derived urinary organic acids that are indicative of metabolic diseases, phenylketonuria, tyrosinemia, and others. Additionally, two-dimensional GC is used for the determination of 2,2,3,3,4,6-hexachlorobiphenyl in milk [67][29]. Food products often contain complex matrix interferences, such as emulsifiers, thickeners, stabilizers, pigments, antioxidants, and others, making it difficult to analyze the analytes of interest. As such, multidimensional GC coupled with infrared or mass spectroscopy may be used to improve the outcomes (Figure 1 and Table 1) [68][30].
Table 1.
Utilization of multidimensional chromatography in food products, biological samples, industrial, environmental, and toxins.
Sample Solvent System Analytes Operation Mode Ref
Fish and chicken Acetonitrile Enrofloxacin SPE–HPLC [69][31]
Pig urine, human plasma, and Smashed shrimp samples Water, methanol, and acetonitrile Bisphenol A C18 SPE–HPLC [70][32]
Wheat, barley, potato, and carrots Acetonitrile and acetic acid Fenuron HPLC–DAD [71][33]
Baby food, chicken meat, vegetables (carrots, tomatoes, green beans, onions, peas, and leeks), potatoes puree, and olive oil Formic acid and acetonitrile Quinolones and fluoroquinolones SAX or MIP cartridges- HPLC [72][34]
Soy samples Acetonitrile Parabens Off-line MISPME by GC-FID [73][35]
Corn Water and Acetonitrile Phenylurea herbicides HPLC–MISPE [74][36]
Pork and chicken Acetonitrile Sulfonamides SPE–HPLC [75][37]
Egg, honey, duck, and lobster samples Methanol: acetonitrile Tetracycline LC-tMS-MISPE [76][38]
Milk Acetic or trichloroacetic acid Chloramphenicol Voltammetry [77][39]
Coffee Acetic or trichloroacetic acid Benzo[a]pyrene HPLC–FLU
Citrus fruits Acetonitrile, sodium dihydrogen Phosphate Thiabendazole CE MISPE [78][40]
Potato, corn, pea, Toluene in acetonitrile Triazines SPE-PIP [79][41]
dried milk, condensed milk, and dried cheese Solvents for solubility Melamine, cyanuric acid DART–TOFMS, LC–MS/MS and ELISA [80][42]
Human growth hormone Ammoniumacetate (AmAc), pH 6.8 solution at 20 mM concentration Reslizumab, bevacizumab EC-IEX-UV-nMS system [81][43]
Peptides mapping tryptic digest D1 mobile phase: (A) 10 Mm

CH3COONH4 in H2O v/v (pH 9); (B) 10 mM

CH3COONH4in H2O/ACN 10:90, v/v (pH 9).

D2 mobile phase: (A) 0.1% TFA in H2O v/v (pH 2); (B) 0.1% TFA in H2O/ACN 10:90, v/v (pH 2).		 Amino acids. α-Casein and dephosphorylated α-casein 2D LC system in which RPLC × RPLC system, coupled to PDA and MS detection. [82][44]
Saccharomyces (yeast protein) Four buffer solutions: Buffer A (2% ACN), buffer B (80% ACN), buffer C (250 mM ammonium acetate/2% ACN), and buffer D (2 M ammonium acetate/2% ACN) with each 0.1% formic acid Soluble, urea-solubilized, and SDS-solubilized proteins RP1-SCX-RP2 microcapillary column and an LCQ Deca XPMS/ Online [83][45]
S. cerevisiae and E. coli Isocratic-acetonitrile and KCl each with 20 mM ammonium formate at pH 10 Tryptic peptides 3-D RP-SAX-RP and RP-RP-LCQ/ Online [84][46]
HEK 293T cell lysates Buffer solutions with various pH 12, 6, 2 with variance 7-35% con Buffer Peptides SAX-based SISPROT/ Off-line [85][47]
Tryptic digest of whole Jurkat cell lysate Water and acetonitrile with gradient system with each 0.1% HFBA Peptides LC-MS/MS (TripleTOF 5600)/Offline [86][48]
Mouse brain cell proteome digest Water and acetonitrile with gradient system with each 0.1% FA and ammonium acetate solution (pH 2.88) Peptides SCX-RPLC-CZE-MS/MS/ Combined [87][49]
Breath of healthy smokers and non-smokers, and patients with COPD - 13 VOC and NSCLC GCMS HP 6890 GC coupled with an HP 5973 mass selective detector [88][50]
Human urine - Aromatic amines GC–MS/MS Agilent 7890-7000 C GC–MS/MS [89][51]
Microwave and conventional oven food contact material - Fatty foods—microwave meals, GC–MS_EI [90][52]
Water (A) and acetonitrile (B), both with 0.1% formic

acid (+), and 0.16 M ammonium hydroxide (-)		 Roasting meat and poultry LC–MS_ESI [90][52]
Tuna fish muscle and wastes - Lipid extract QqQ MS, [91][53]
Tuna β-actin 0.1% TFA in water and 0.1% TFA in water: ACN Peptides RP-UPLC with ESI-ion trap-TOF MS/MS [91][53]

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