Separation Of Amino Acids Research Articles

Sitetack: A deep learning model that improves PTM prediction by using known PTMs.

Post-translational modifications (PTMs) increase the diversity of the proteome and are vital to organismal life and therapeutic strategies. Deep learning has been used to predict PTM locations. Still, limitations in datasets and their analyses compromise success. We evaluated the use of known PTM sites in prediction via sequence-based deep learning algorithms. For each PTM, known locations of that PTM were encoded as a separate amino acid before sequences were encoded via word embedding and passed into a convolutional neural network that predicts the probability of that PTM at a given site. Without labeling known PTMs, our models are on par with others. With labeling, however, we improved significantly upon extant models. Moreover, knowing PTM locations can increase the predictability of a different PTM. Our findings highlight the importance of PTMs for the installation of additional PTMs. We anticipate that including known PTM locations will enhance the performance of other proteomic machine learning algorithms. Sitetack is available as a web tool at https://sitetack.net; the source code, representative datasets, instructions for local use, and select models are available at https://github.com/clair-gutierrez/sitetack. Supplementary data are available at Bioinformatics online.

Comprehensive review on ethnomedicinal, phytochemistry and pharmacological profile of.

Ficus religiosa L. (F. religiosa) or sacred fig is a large perennial tree belonging to the family Moraceae or mulberry family. Though the tree has pan-tropical distribution but originally it is indigenous to the Indian subcontinent and Indochina region. Popularly the tree is named "Pepal or bodhi tree". Traditionally, it is practiced for the treatment of asthma, nose bleeding, heart disorders, diabetes, wound healing, ear problems, constipation, hyperlipidemia, gonorrhea, ulcers and infectious disorders. Chemical analysis demonstrated the presence of numerous bioactives including tannins, phenols, saponins, sugars, alkaloids, methionine, terpenoids, flavonoids, glycosides, proteins, separated amino acids, essential and volatile oils and steroids etc., which are probably responsible for its diverse pharmacological actions. The present work is an attempt to compile up-to-date comprehensive information on F. religiosa that covers its taxonomy, ethnomedicinal importance, phytochemistry, pharmacological attributes and clinical trials. Keeping in mind the various health attributes of F. religiosa, future research can be aimed at in-depth elucidation of the structure-function relationship and multifactorial signalings pathways.

Highly efficient chiral extraction of amino acid enantiomers using Wei-Phos-Pd as chiral extractant

The development of novel and highly efficient chiral extractants has always been a research hotspot in the field of chiral extraction. In this work, Wei-Phos, an adaptive chiral sulfinamide phosphine ligand featuring multiple coordination centers, was employed as a chiral extractant to separate amino acid enantiomers. The influences of metal precursors, organic solvents, pH of the aqueous phase, extractant concentrations, and extraction temperature on enantioselectivities were investigated. The results show that Wei-Phos-Pd is a highly efficient chiral extractant for amino acid enantiomers. D-amino acids are preferentially recognized by Wei-Phos-Pd. The separation factors for phenylalanine (Phe), homophenylalanine (Hph), 3-chlorophenylglycine (3-Cl-Phg), and 4-nitro-phenylalanine (4-NO2-Phe) are 12.19, 8.89, 13.17 and 7.06, respectively. The structure–activity study and recognition mechanism analysis indicate that the chiral tert-butylsulfinamide group and diphosphine structure in Wei-Phos have significant influences on enantioselectivities. The multiple coordination centers account for the high enantioselectivities of Wei-Phos. Wei-Phos-Pd stands out as the most efficient and versatile chiral diphosphine-metal extractant for chiral extraction of amino acids to date.

MAGPIE: An interactive tool for visualizing and analyzing protein-ligand interactions.

Quantitative tools to compile and analyze biomolecular interactions among chemically diverse binding partners would improve therapeutic design and aid in studying molecular evolution. Here we present Mapping Areas of Genetic Parsimony In Epitopes (MAGPIE), a publicly available software package for simultaneously visualizing and analyzing thousands of interactions between a single protein or small molecule ligand (the "target") and all of its protein binding partners ("binders"). MAGPIE generates an interactive three-dimensional visualization from a set of protein complex structures that share the target ligand, as well as sequence logo-style amino acid frequency graphs that show all the amino acids from the set of protein binders that interact with user-defined target ligand positions or chemical groups. MAGPIE highlights all the salt bridge and hydrogen bond interactions made by the target in the visualization and as separate amino acid frequency graphs. Finally, MAGPIE collates the most common target-binder interactions as a list of "hotspots," which can be used to analyze trends or guide the de novo design of protein binders. As an example of the utility of the program, we used MAGPIE to probe how different antibody fragments bind a viral antigen; how a common metabolite binds diverse protein partners; and how two ligands bind orthologs of a well-conserved glycolytic enzyme for a detailed understanding of evolutionarily conserved interactions involved in its activation and inhibition. MAGPIE is implemented in Python 3 and freely available at https://github.com/glasgowlab/MAGPIE, along with sample datasets, usage examples, and helper scripts to prepare input structures.

Sitetack: A Deep Learning Model that Improves PTM Prediction by Using Known PTMs.

Post-translational modifications (PTMs) increase the diversity of the proteome and are vital to organismal life and therapeutic strategies. Deep learning has been used to predict PTM locations. Still, limitations in datasets and their analyses compromise success. Here we evaluate the use of known PTM sites in prediction via sequence-based deep learning algorithms. Specifically, PTM locations were encoded as a separate amino acid before sequences were encoded via word embedding and passed into a convolutional neural network that predicts the probability of a modification at a given site. Without labeling known PTMs, our model is on par with others. With labeling, however, we improved significantly upon extant models. Moreover, knowing PTM locations can increase the predictability of a different PTM. Our findings highlight the importance of PTMs for the installation of additional PTMs. We anticipate that including known PTM locations will enhance the performance of other proteomic machine learning algorithms.

Leucine zipper as a bridge for transaminase self-assembly: A fusion enzyme for efficient chiral conversion of d-phenylglycine

Amino acid transferase is a family of enzymes used to catalyze and separate chiral amino acids. However, due to the low efficiency, by-products and reverse reactions occur in cascade reactions. Therefore, in the research, phenylglycine aminotransferase and aspartate aminotransferase were self-assembled in vitro by leucine zipper. The self-assembled enzyme system with d-phenylglycine and α-ketoglutarate as substrates were used for the chiral transformation reaction. By studying the enzyme combination, kinetic reaction stability and catalytic efficiency, it was found that the self-assembled enzyme showed improved stability and better affinity to the substrate than the control and achieved only ee value of 17.86% for the control at the substrate ratio was 1:2. In contrast, the self-assembled enzyme basically catalyzed the complete conversion of d-Phg to l-Phg, with the ee value as 99%. These results demonstrated the feasibility of the leucine zipper and the conversion of d-phenylglycine to the l-type by fusion enzyme.

Click preparation of triazole-bridged teicoplanin-bound chiral stationary phases for efficient separating amino acid enantiomers

Enantioseparation of amino acids is considered as a challenging task due to the extreme structural similarity of their enantiomers. Herein, teicoplanin was modified with different chemical equivalents of azide groups and attached to silica particles by employing Click Chemistry for resolution of chiral amino acids for the first time. Interestingly, teicoplanin modified with 5-fold the chemical equivalent of azide groups (TK-2 CSP) exhibited superior amino acid separation ability compared to two other columns: one modified with only 1-fold the chemical equivalent of azide groups (TK-1 CSP), and the other modified with excess azide groups (TK-3 CSP). Additionally, the TK-2 CSP exhibited superior enantioselectivity when separating amino acids containing hydrophobic alkyl side chains in comparison to other teicoplanin-based CSPs. The TK-2 CSP column allows the baseline separation of 7 native amino acids. Molecular docking demonstrates that effective enantioseparation arises from distinct patterns of interaction between the host and guest molecules. Moreover, (p-methyl) phenylcarbaminoylated-teicoplanin CSP (TK-4, TK-5 CSP) were prepared by post-modification from TK-1 CSP and TK-2 CSP to isolate Fmoc-modified amino acids. This work explores the impact of various modification methods on the enantioseparation effects of host molecules and paves the way for expanding the potential applications of teicoplanin and macrocyclic glycopeptide molecules.

Electrochemical-assisted synthesis of molecularly imprinted graphene oxide/magnetite for highly selective enantiomer separation

Enantiomeric compounds have been reported to exhibit different biological and pharmacological activities. MIP-GO/Fe3O4 has been synthesized with L-glutamic acid (L-Glu) as template molecules and developed to separate amino acids enantiomeric compounds (AAs) especially for DL-glutamic acid (DL-Glu). The MIP-GO/Fe3O4 adsorbent material was synthesized in several stages, namely electrochemically synthesizing GO, co-precipitation for GO/Fe3O4, and surface imprinting polymerization to obtain MIP-GO/Fe3O4. The characterization results of MIP-GO/Fe3O4 show that the composite system has been successfully created with saturation magnetization (Ms) values of 49.37 emu/g. Adsorption tests are carried out on the racemic mixture of DL-Glu and show that the material worked effectively and selectively to separate D- and L-Glu with a %ee value of 97.86 % and an adsorption capacity for L of 9.9 mg g−1 on optimum conditions. In addition, the adsorbent developed shows good reusability values with a decreasing performance of no more than 2.0 % after 10-time use cycles and is easily separated in less than 15 s. This research contributes to developing advanced materials with extraordinary selectivity and efficiency, paving the way for advances in enantiomer separation technology.

Chiral zwitterionic stationary phases based on Cinchona alkaloids and dipeptides – design, synthesis and application in chiral separation

Chiral resolution of polar organic compounds such as amino acids and peptides represents an important chromatographic task due to increasing significance of natural species, which play important signaling and regulatory roles in the living organisms. Despite the number of available chiral stationary phases, this task remains challenging, since not many of the commercially available systems are capable to resolve non-derivatized zwitterionic species. In this study, we present a target-oriented design of a new class of chiral selectors. Pursuing the goal to separate amino acids, and especially short peptides, we have combined Cinchona alkaloids – quinine and quinidine – with three different biogenic dipeptides. We have synthesized six different chiral stationary phases, with selector loading of ∼200 μmol g−1, and tested their chiral recognition capabilities for acidic, basic and zwitterionic analytes using various mobile phases. We have observed that all chiral stationary phases retain the chiral anion exchange capability known for commercially available Cinchona-based columns leading to baseline or partial resolution of six out of ten analytes. The performance in chiral resolution of basic analytes is not optimum due to the weak cation exchange character of the peptidic residue. However, we report on encouraging results in the chiral resolution of short peptides, for which, depending on their structure, we see the chiral resolution of up to three stereoisomers (from four possible) in a preliminary screening.

Non-Enzymatic Formation of N-acetylated Amino Acid Conjugates in Urine

Unknown N-acylated amino acid (N-AAA) conjugates have been detected in maple syrup urine disease (MSUD) and other inborn errors of metabolism (IEMs). This study aimed to elucidate the mechanism behind the formation of urinary N-AAA conjugates. Liquid–liquid extraction was employed to determine the enantiomeric composition of N-AAA conjugates, followed by liberation of conjugated amino acids through acid hydrolysis. Gas chromatography–mass spectrometry (GC–MS) was used to separate amino acid enantiomers. In vitro experiments were conducted to test the non-enzymatic formation of N-AAA conjugates from 2-keto acids and ammonia, with molecular modelling used to assess possible reaction mechanisms. Adequate amounts of N-AAA conjugates were obtained via organic acid extraction without concurrent extraction of native amino acids, and hydrolysis was complete without significant racemisation. GC–MS analysis successfully distinguished amino acid enantiomers, with some limitations observed for L-isoleucine and D-alloisoleucine. Furthermore, investigation of racemic N-AAA conjugates from an MSUD case confirmed its non-enzymatic origin. These findings highlight the value of employing chiral strategy and molecular modelling to investigate the origin of unknown constituents in biological samples. Additionally, these conjugates warrant further investigation as potential factors contributing to MSUD and other IEMs.

Variation among arthropod taxa in the amino acid content of exoskeleton and digestible tissue.

Arthropod consumption provides amino acids to invertebrates and vertebrates alike, but not all amino acids in arthropods may be digestible as some are bound in the exoskeleton. Consumers may not be able to digest exoskeleton in significant amounts or avoid it entirely (e.g., extraoral digestion). Hence, measures that do not separate digestible amino acids from those in exoskeleton may not accurately represent the amino acids available to consumers. Additionally, arthropods are taxonomically diverse, and it remains unclear if taxonomic differences also reflect differences in amino acid availability. Thus, we tested: (1) if there were consistent differences in the content and balance of amino acids between the digestible tissue and exoskeleton of arthropods and (2) if arthropod Orders differ in amino acid content and balance. We measured the amino acid content (mg/100 mg dry mass) and balance (mg/100 mg protein) of whole bodies and exoskeleton of a variety of arthropods using acid hydrolysis. Overall, there was higher amino acid content in digestible tissue. There were also significant differences in the amino acid balance of proteins in digestible tissue and exoskeleton. Amino acid content and balance also varied among Orders; digestible tissues of Hemiptera contained more of some essential amino acids than other Orders. These results demonstrate that arthropod taxa vary in amino acid content, which could have implications for prey choice by insectivores. In addition, exoskeleton and digestible tissue content differ in arthropods, which means that whole body amino acid content of an arthropod is not necessarily a predictor of amino acid intake of a predator that feeds on that arthropod.

Aptamer-Protein Structures Guide In Silico and Experimental Discovery of Aptamer-Short Peptide Recognition Complexes or Aptamer-Amino Acid Cluster Complexes.

A method to computationally and experimentally identify aptamers against short peptides or amino acid clusters is introduced. The method involves the selection of a well-defined protein aptamer complex and the extraction of the peptide sequence participating in the binding of the protein to the aptamer. The subsequent fragmentation of the peptide sequence into short peptides and the in silico docking-guided identification of affinity complexes between the miniaturized peptides and the antiprotein aptamer, followed by experimental validation of the binding features of the short peptides with the antiprotein aptamers, leads to the identification of new short peptide-aptamer complexes. This is exemplified with the identification of the pentapeptide RYERN as the scaffold that binds thrombin to the DNA thrombin aptamer (DNA TA). In silico docking studies followed by microscale thermophoresis (MST) experiments demonstrate that the miniaturized tripeptides RYE, YER, and ERN reveal selective binding affinities toward the DNA TA. In addition, docking and MST experiments show that the ribonucleotide-translated RNA TA shows related binding affinities of YER to the DNA TA. Most importantly, we demonstrate that the separated amino acids Y/E/R assemble as a three amino acid cluster on the DNA TA and RNA TA aptamers in spatial configurations similar to the tripeptide YER on the respective aptamers. The clustering phenomenon is selective for the YER tripeptide system. The method to identify binding affinities of miniaturized peptides to known antiprotein aptamers and the specific clustering of single amino acids on the aptamers is further demonstrated by in silico and experimental identification of the binding of the tripeptide RET and the selective clustering of the separated amino acids R/E/T onto a derivative of the AS1411 aptamer against the nucleolin receptor protein.

Expansion of the genetic code through reassignment of redundant sense codons using fully modified tRNA.

Breaking codon degeneracy for the introduction of non-canonical amino acids offers many opportunities in synthetic biology. Yet, despite the existence of 64 codons, the code has only been expanded to 25amino acids in vitro. A limiting factor could be the over-reliance on synthetic tRNAs which lack the post-transcriptional modifications that improve translational fidelity. To determine whether modified, wild-type tRNA could improve sense codon reassignment, we developed a new fluorous method for tRNA capture and applied it to the isolation of roughly half of the Escherichia coli tRNA isoacceptors. We then performed codon competition experiments between the five captured wild-type leucyl-tRNAs and their synthetic counterparts, revealing a strong preference for wild-type tRNA in an in vitro translation system. Finally, we compared the ability of wild-type and synthetic leucyl-tRNA to break the degeneracy of the leucine codon box, showing that only captured wild-type tRNAs are discriminated with enough fidelity to accurately split the leucine codon box for the encoding of three separate amino acids. Wild-type tRNAs are therefore enabling reagents for maximizing the reassignment potential of the genetic code.

Diagnosing Metabolic Diseases Using Thin Layer Chromatography: Laboratory Experiment for both Organic Chemistry and Biochemistry Undergraduate Laboratories

With the ability to resolve metabolites as well as proteins, Thin Layer Chromatography (TLC) is a technique that is used in both Organic Chemistry as well as Biochemistry classes. There were two goals of this project: first was to develop a virtual laboratory experience for students in Organic Chemistry lab that expanded upon a previous experiment with TLC. The second was a project in method development for Biochemistry students. Students in Organic Chemistry lab had previously completed an experiment in TLC that required identification of an analgesic by comparing Rf value to those of standards. This project produced a virtual one where students were expected to diagnose various diseases based on TLC of affected amino acids. The diseases involved were Phenylketonuria, ornithine transcarbamylase (OTC) deficiency, arginemia, tyrosinemia, maple syrup urine disease citrullinemia type I and type II which respectively resulted in increased levels of amino acids phenylalanine, ornithine, arginine, tyrosine, either leucine, isoleucine or valine, or citrulline. The procedure involved spotting the standards and “plasma from blood” samples on the TLC plate with 70/30 mixture of n‐propanol/water. Amino acids were visualized by staining with ninhydrin. Several of the relevant amino acids have Rf values that are similar. In order to correctly identify the disease, a second piece of information was needed: the relative level of urea. Four of the diseases are urea cycle disorders that produce decreased levels of urea while the other three have normal levels. Students were provided with a picture of a standard curve of urea as well as pictures of the patients; this was used to identify if the disease was a UCD or not. In order to assess learning, students were given a pre and post test assessment. Students had previously done a TLC and 73.8% were able to correctly identify the role of various components when given a copy of the procedure. There was a 5.8% increase in score in the post test. For questions that addressed the theory of how TLC works or other applications, 37.7% of students on pre test could answer correctly. This value increased to 55.0% on the post test; this result suggests were able to deepen their knowledge of TLC with a second exposure. Another approach was taken for the Biochemistry students. Method development has been shown to be a critical skill, yet it is difficult to teach. Biochemistry students were provided with “blood” of five types of patients (normal, arginemia, OTC deficiency, citrullinemia type I and II), basic procedure for separating amino acids on TLC, and the amounts of these amino acids normally found in blood as well as in the disease state. The question for these students was if it was possible to detect these diseases using TLC? Students were given a pre test assessment. The great majority (81.2%) were able to partially or completely identify the function of the components in TLC. Students were asked to list questions that must be addressed in order to answer this question when provided with blood or plasma. Student responses covered a wide range and addressed both valid questions (do the amino acids have different Rfvalues or is the amount of amino acid detectable in this method) as well as not valid questions to having no answers.

Interactions of productivity, content of separate amino acids, general protein in blood serum of chickens and tone of the autonomic nervous system

The regulation of metabolic processes and maintenance of a stable internal environment of the body is the role of the autonomic nervous system (ANS). It influences all systems and organs, thus along with the neuro-humoral system providing control over the metabolism of proteins, fats and carbohydrates. Amino acids, as structural units of any protein molecule, are essential for the metabolism of proteins, enzymes and hormones. Certain amino acids are involved in neutralizing free radicals and are a donor of metal groups, can act as an energy source in case of a deficiency of carbohydrates and fats, and be structural units of connective tissue. Exchange of these compounds and their regulation by ANS have not been studied sufficiently, therefore, it is necessary to carry out research in this direction. The studies were aimed at finding the presence of the regulatory effect of ANS on the content of individual amino acids in the blood serum of 60-day-old Cobb-500 chickens. Determination of the ANS tone in Cobb-500 chickens at the age of 30–35 days was determined by the method of variation pulsometry. Electrocardiographic examination was performed in a quiet room without the use of sedatives. At the age of 60 days, blood was obtained from the saphenous vein of the chickens’ shoulder. Blood was sampled from 10 am to 1 pm, after a short fasting (2:00). It was found that sympathicotonic chickens had a higher content of individual amino acids compared to normotonics and vagotonics. In vagotonic chickens, the content of glycine and serine in the blood serum significantly exceeded the rate of chickens with a balanced tone of ANS. Correlative relationships between amino acid content were highly significant in poultry with balanced ANS tone. Vagotonic chickens had the highest correlations between ANS tone, heart rate and amino acid content (trend). Correlative relationships were present between ANS and body weight of chickens of different groups, serum protein. The correlation between total serum protein and body weight of chickens was different depending on the tone of the ANS. Fisher’s test revealed a significant effect of balanced ANS tone on the content of valine, serine and glycine and the weight of chickens. The effect of ANS tone on weight in vagotonic birds was high.

Layered graphene oxide membranes functioned by amino acids for efficient separation of metal ions

As a kind of heavy metal, lead ions (Pb2+) are a great threat to human health and the ecological environment. Due to their excellent properties, graphene and its derivatives are expected to be used for water purification. Amino acid functional groups were designed for graphene oxide membranes via dehydration condensation. The effects of the amino acid groups on the membrane performance for separating metal ions were investigated by the molecular dynamics. Under the premise that the water permeability is basically the same, the retention rate for Pb2+ by alanine groups with negatively charged terminals is 1.6 times of that by the carboxyl groups. The number-density map and the radial distribution function indicate that a large area of local high-density regions of Pb2+ will be formed around the negatively charged alanine groups. At the same time, the diffusion coefficient for Pb2+ and Na+ can be effectively reduced by the electrostatic attraction of the negative groups. It is concluded that functioned by the negatively charged alanine, the retention rate for metal ions by the layered graphene oxide membranes can be significantly improved at the cost of a little loss in water permeability.

Preparation and evaluation a mixed-mode stationary phase with C18 and 2-methylindole for HPLC.

A modified C18 column (Silpr-2MI-C18) was prepared using 2-methylindole and C18 reagent. The extent of C18 hydrocarbon chain, conjugative rings and anion exchange site provided multiple retention mechanisms, including reversed-phase liquid chromatography (RPLC), π-π interaction, hydrophilic interaction liquid chromatography (HILIC) and anion exchange chromatography (AEC). The separation of protected amino acids was investigated on the commercial C18 and Silpr-2MI-C18 columns, while the chromatographic conditions, including methanol content and pH of the mobile phase, were studied. The separation arrangement of the hydrophilic amino acids was different on the Silpr-2MI-C18 column compared to the commercial C18 column under RPLC mode. Furthermore, these amino acids were separated on the Silpr-2MI-C18 column under HILIC mode. The modified C18 column was employed to separate amino acids, alkylbenzenes and polycyclic aromatic hydrocarbons under RPLC mode and inorganic anion under AEC mode. The results confirm that this new stationary phase of RPLC/HILIC/AEC has multiple interactions with different analytes. Effective retention of biological samples was found on the Silpr-2MI-C18 column by comparing the results obtained from the commercial C18 column.

Distinct interactions of eIF4A and eIF4E with RNA helicase Ded1 stimulate translation in vivo.

Yeast DEAD-box helicase Ded1 stimulates translation initiation, particularly of mRNAs with structured 5'UTRs. Interactions of the Ded1 N-terminal domain (NTD) with eIF4A, and Ded1-CTD with eIF4G, subunits of eIF4F, enhance Ded1 unwinding activity and stimulation of preinitiation complex (PIC) assembly in vitro. However, the importance of these interactions, and of Ded1-eIF4E association, in vivo were poorly understood. We identified separate amino acid clusters in the Ded1-NTD required for binding to eIF4A or eIF4E in vitro. Disrupting each cluster selectively impairs native Ded1 association with eIF4A or eIF4E, and reduces cell growth, polysome assembly, and translation of reporter mRNAs with structured 5'UTRs. It also impairs Ded1 stimulation of PIC assembly on a structured mRNA in vitro. Ablating Ded1 interactions with eIF4A/eIF4E unveiled a requirement for the Ded1-CTD for robust initiation. Thus, Ded1 function in vivo is stimulated by independent interactions of its NTD with eIF4E and eIF4A, and its CTD with eIF4G.

Liquid Chromatographic Isolation of Individual Amino Acids Extracted From Sediments for Radiocarbon Analysis

The “building blocks of life” are found nearly ubiquitously in the environment in the form of proteins, peptides, and single amino acids. To shed light on amino acid sources, cycling, and preservation in sedimentary environments, we present a method using high-pressure liquid chromatography to separate and isolate underivatized amino acids extracted from sediments to conduct compound-specific radiocarbon analysis. This method consists of three main steps including (1) amino acid extraction by hydrofluoric and hydrochloric acids followed by desalting, (2) liquid chromatographic isolation and purification using two complementary column chemistries, and (3) post-purification and measurement by accelerator mass spectrometry. The resulting blank of this procedure is estimated to contain 2.2±1.3 µgC with 0.25±0.09 Fm.

A click tyrosine zwitterionic stationary phases for hydrophilic interaction liquid chromatography

New zwitterionic (ZIC) stationary phases (SPs) are synthesized with the click and conventional bonding of tyrosine to silica gel. Infrared spectra and elemental analysis demonstrate the successful click and conventional bonding of this ZIC group on silica particles by the surface coverage including 2.36 and 0.75 µm m−2, respectively. Given the above-mentioned explanation, the present study evaluated the retention mechanism and chromatographic manners of polar compounds on these new materials under hydrophilic interaction liquid chromatography (HILIC) conditions. Based on the results, the Click-Tyrosine Stationary Phase provided good HILIC characteristics when it was applied to separate phenolic compounds, amino acids, alkaloids, and nucleobases compared to bare silica gel SP and even conventional tyrosine SPs. Further, this new Click-Tyrosine-SP represented appropriate HILIC features and column efficiency (the theoretical plate number was up to 50,000 plates m−1 for thebaine). Furthermore, the study investigated the effect of solute polarity (the number of the hydroxyl group of phenolic compounds) and hydrophobicity (the number of the side chain of aliphatic amino acids) on retention behaviors. Finally, some important factors were studied as the potential variables for guiding the retention behavior of the polar compound in HILIC condition including solvent composition, salt concentration, and the buffer pH of the mobile phase.