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Fulltext Complete genome sequencing of Pandoraea pnomenusa RB38 and Molecular Characterization of Its N-acyl homoserine lactone synthase gene ppnI

Lim YL, Ee R, How KY, Lee SK, Yong D, Tee KK, et al.

PeerJ, 2015;3:e1225.
PMID: 26336650 DOI: 10.7717/peerj.1225

In this study, we sequenced the genome of Pandoraea pnomenusa RB38 using Pacific Biosciences RSII (PacBio) Single Molecule Real Time (SMRT) sequencing technology. A pair of cognate luxI/R homologs was identified where the luxI homolog, ppnI, was found adjacent to a luxR homolog, ppnR1. An additional orphan luxR homolog, ppnR2, was also discovered. Multiple sequence alignment and phylogenetic analysis revealed that ppnI is an N-acyl homoserine lactone (AHL) synthase gene that is distinct from those of the nearest phylogenetic neighbor viz. Burkholderia spp. High resolution tandem mass spectrometry (LC-MS/MS) analysis showed that Escherichia coli BL21 harboring ppnI produced a similar AHL profile (N-octanoylhomoserine lactone, C8-HSL) as P. pnomenusa RB38, the wild-type donor strain, confirming that PpnI directed the synthesis of AHL in P. pnomenusa RB38. To our knowledge, this is the first documentation of the luxI/R homologs of the genus Pandoraea.

Matched MeSH terms: Chromatography, Liquid
Fluoride adsorption by doped and un-doped magnetic ferrites CuCe(x)Fe(2-x)O4: Preparation, characterization, optimization and modeling for effectual remediation technologies

Rehman MA, Yusoff I, Alias Y

J Hazard Mater, 2015 Dec 15;299:316-24.
PMID: 26143194 DOI: 10.1016/j.jhazmat.2015.06.030

A series of doped and un-doped magnetic adsorbents CuCexFe2-xO4 (x=0.0-0.5) for fluoride were prepared with the micro-emulsion method. Fluoride adsorption was optimized for solution pH, temperature, contact time, and initial concentration and was monitored via normal phase ion chromatography (IC). The effect of concomitant anions was also explored to perform and simulate competitive fluoride adsorption in real water samples. Optimal adsorption was discovered by a simple quadratic model based on central composite design (CCD) and the response surface method (RSM). The adsorption, electrochemical and magnetic properties were compared between doped and un-doped ferrites. Doped ferrites (x=0.1-0.5) were found to be superior to un-doped ferrites (x=0) regarding the active sites, functional groups and fluoride adsorption. The characterization, optimization and application results of the doped ferrites indicated enhanced fluoride adsorption and easy separation with a simple magnet.

Matched MeSH terms: Chromatography
Fulltext Isolation, Characterization, Crystal Structure Elucidation of Two Flavanones and Simultaneous RP-HPLC Determination of Five Major Compounds from Syzygium campanulatum Korth

Memon AH, Ismail Z, Al-Suede FS, Aisha AF, Hamil MS, Saeed MA, et al.

Molecules, 2015;20(8):14212-33.
PMID: 26248073 DOI: 10.3390/molecules200814212

Two flavanones named (2S)-7-Hydroxy-5-methoxy-6,8-dimethyl flavanone (1), (S)-5,7-dihydroxy-6,8-dimethyl-flavanone (2), along with known chalcone, namely, (E)-2',4'- dihydroxy-6'-methoxy-3',5'-dimethylchalcone (3) and two triterpenoids, namely, betulinic and ursolic acids (4 and 5), were isolated from the leaves of Syzygium campanulatum Korth (Myrtaceae). The structures of compounds (1 and 2) were determined on the basis of UV-visible, FTIR, NMR spectroscopies and LC-EIMS analytical techniques. Furthermore, new, simple, precise, selective, accurate, highly sensitive, efficient and reproducible RP-HPLC method was developed and validated for the quantitative analysis of the compounds (1-5) from S. campanulatum plants of five different age. RP-HPLC method was validated in terms of specificity, linearity (r2 ≤ 0.999), precision (2.0% RSD), and recoveries (94.4%-105%). The LOD and LOQ of these compounds ranged from 0.13-0.38 and 0.10-2.23 μg·mL-1, OPEN ACCESS respectively. Anti-proliferative activity of isolated flavanones (1 and 2) and standardized extract of S. campanulatum was evaluated on human colon cancer (HCT 116) cell line. Compounds (1 and 2) and extract revealed potent and dose-dependent activity with IC50 67.6, 132.9 and 93.4 μg·mL-1, respectively. To the best of our knowledge, this is the first study on isolation, characterization, X-ray crystallographic analysis of compounds (1 and 2) and simultaneous RP-HPLC determination of five major compounds (1-5) from different age of S. campanulatum plants.

Matched MeSH terms: Chromatography, High Pressure Liquid/methods*; Chromatography, Reverse-Phase/methods*
Method development for determination of fluroxypyr in water

Halimah M, Tan YA, Aini K, Ismail BS

J Environ Sci Health B, 2003 Jul;38(4):429-40.
PMID: 12856925

Improved methods for extraction and clean up of fluroxypyr residue in water have been established. Two methods of fluroxypyr extraction were used, namely, Direct Measurement of fluroxypyr and Concentration of fluroxypyr onto A Solid Phase Extraction (SPE) Adsorbent, followed by elution with solvent before determination of fluroxypyr. The recovery for Direct Measurement of fluroxypyr in water containing 8-100 microg L(-1), ranged from 86 to 110% with relative standard deviation of 0.7 to 2.15%. For the second method, three types of SPE were used, viz. C18, C18 end-capped and polyvinyl dibenzene (ISOLUTE ENV+). The procedure involved concentrating the analyte from fluroxypyr-spiked water at pH 3, followed by elution of the analyte with 4 mL of acentonitrile. The recovery of fluroxypyr from the spiked sample at 1 to 50 microg L(-1) after eluting through either C18 or C18 end-capped ranged from 40-64% (with relative standard deviation of 0.7 to 2.15) and 41-65% (with standard deviation of 1.52 to 11.9). The use of ISOLUTE ENV+, gave better results than the C18, C18 end-capped or the Direct Measurement Methods. The recovery and standard deviation of fluroxypyr from spiked water using ISOLUTE ENV+ ranged from 91-102% and 2.5 to 5.3, respectively.

Matched MeSH terms: Chromatography, High Pressure Liquid/methods*; Gas Chromatography-Mass Spectrometry/methods*
Simple high-performance liquid chromatographic method for simultaneous determination of fusidic Acid and betamethasone dipropionate in a cream formulation

Peh KK, Tan YT

Int J Pharm Compd, 2000 May-Jun;4(3):229-31.
PMID: 23986007

A simple and selective high-performance liquid chromatography (HPLC) method using ultraviolet detection was developed for simultaneous determination of fusidic acid and betamethasone dipropionate in a cream formulation. A Supelcosil LC18 column was used for chromatographic separation. The mobile phase consisted of acetonitrile and 0.01 M disodium hydrogen orthophosphate (70:30, % v/v) adjusted to pH 6 with glacial acetic acid. Analysis was run at a flow rate of 1.0 mL/minute with the detector operating at 235 nm. The standard calibration curve was linear over a concentration range of 0.3 to 1.2 mg/mL for fusidic acid and 9.6 to 38.4 micrograms/mL for betamethasone dipropionate. The average recovery values for fusidic acid and betamethasone dipropionate were almost 100%. The within-run and between-run coefficient of variation and percent error values for the two drugs were all less than 2% and +/- 3%, respectively.

Matched MeSH terms: Chromatography, High Pressure Liquid
Survey of the Anopheles maculatus complex (Diptera: Culicidae) in peninsular Malaysia by analysis of cuticular lipids

Kittayapong P, Clark JM, Edman JD, Lavine BK, Marion JR, Brooks M

J Med Entomol, 1993 Nov;30(6):969-74.
PMID: 8271255

Anopheles maculatus Theobald sensu lato is a species complex now consisting of eight sibling species; An. maculatus is still represented by two cytologically distinct forms; i.e., the widely distributed sensu strictu or B, and E from southern Thailand and adjacent areas in northern Malaysia. Cuticular lipid profiles in conjunction with principal component analysis was used to separate An. maculatus form E from sensu stricto form B in a preliminary survey of the An. maculatus complex at five locations spanning peninsular Malaysia. The relative rank orders, from the areas of the five gas chromatographic peaks used to determine lipid differences for specimens from peninsular Malaysia, matched well with those from cytogenetically identified colony specimens of An. maculatus forms B and E. The two-dimensional principal component pattern of specimens identified as form E was highly clumped, which indicated that very similar cuticular lipids were present within this putative malaria vector. Both forms coexisted in peninsular Malaysia, but form E may be dominant except in the south.

Matched MeSH terms: Chromatography, Gas
High performance liquid chromatography (HPLC) method for confirming thin layer chromatography (TLC) findings in inborn errors of metabolism children in Malaysia

Yahya NA, Ismail Z, Embong KH, Mohamad SA

Southeast Asian J. Trop. Med. Public Health, 1995;26 Suppl 1:130-3.
PMID: 8629091

High performance liquid chromatography (HPLC) with phenylisothiocyanate (PITC) is recently used for confirming the diagnosis of inborn errors of metabolism (IEM) especially amino acid disorders in Malaysian children. The method of HPLC used is a precolumn derivatization of amino acids with phenylisothiocyanate and is separated by reversed phase chromatography using 3.9 x 300 mm free amino acid columns and is detected by a UV/Vis detector. The samples are obtained from cases suspected of inborn errors of metabolism, especially of amino acid disorders, which are detected clinically by pediatricians. Initially, samples from patients suspected of inborn errors of metabolism, either urine or serum, are run on one-dimensional thin layer chromatography and supplementary chemical tests to detect the abnormal bands and associated abnormalities respectively. Positive samples are further run on HPLC to determine the specific amino acids abnormality. An examples of a case of maple syrup urine disease is discussed, based on the thin layer chromatography findings and HPLC findings.

Matched MeSH terms: Chromatography, High Pressure Liquid/methods; Chromatography, Thin Layer/methods
The fatty acid composition of cooking oils and fats used in Malaya

Chong YH, Mills GL

Med J Malaya, 1966 Dec;21(2):125-7.
PMID: 4227381
Matched MeSH terms: Chromatography, Gas
HPLC-Based Qualitative and Quantitative Analyses of Alkaloids in Chewable Areca Products from Different Geographic Regions

Cao M, Liu Y, Yuan H, Qiu Y, Xie Q, Yi P, et al.

J AOAC Int, 2020 Sep 01;103(5):1400-1405.
PMID: 33241395 DOI: 10.1093/jaoacint/qsaa048

BACKGROUND: Chewable areca products are popular in Asian countries, including India, Pakistan, Malaysia, and China. The major alkaloids present in areca products are guvacine, arecaidine, guvacoline, and arecoline which cause carcinogenicity and addiction.
OBJECTIVE: The objective of this study was the quantitative analysis of the alkaloid content of areca chewable products from different countries and regions using HPLC-UV, as well as the benefit of their safety evaluation products.
METHOD: An HPLC-UV method was established for qualitative and quantitative analyses of 65 batches of areca chewable products from different countries and regions. Additionally, similarity evaluation of chromatographic fingerprints was applied for data analysis.
RESULTS: These results reveal a significant variation in the levels of areca alkaloids among tested products, specifically guvacoline (0.060-1.216 mg/g), arecoline (0.376-3.592 mg/g), guvacine (0.028-1.184 mg/g), and arecaidine (0.184-1.291 mg/g). There were significant differences in the alkaloid content of areca chewable products from different producing areas.
CONCLUSIONS: The method will be useful in the safety evaluation of different areca chewable products.
HIGHLIGHTS: The established HPLC-UV method can be adopted for safety evaluation of areca chewable products from different countries and regions due to its general applicability.

Matched MeSH terms: Chromatography, High Pressure Liquid
Fulltext Thymoquinone content in marketed black seed oil in Malaysia

Alkhatib H, Mawazi SM, Al-Mahmood SMA, Zaiter A, Doolaanea AA

J Pharm Bioallied Sci, 2020 07 18;12(3):284-288.
PMID: 33100788 DOI: 10.4103/jpbs.JPBS_208_20

Thymoquinone (TQ) is the major active compound in black seed oil (BSO). Many pharmacological effects of TQ, such as anti-inflammatory, hypoglycemic, antioxidant, immune stimulator, and anticancer, have been reported. TQ can be considered as a biomarker for BSO, but its content in the commercial products is rarely reported. TQ content varies based on the oil source and extraction method. This study aimed to quantify the TQ content in the commercial BSO products in Malaysia and to evaluate whether the products can be used as a source of TQ for therapeutic benefits. TQ was quantified using an established high-performance liquid chromatography (HPLC) method. TQ human equivalent dose (HED) was calculated based on reported animal studies from literature, and theoretical BSO amount containing the TQ dose was calculated based on the HPLC analysis. TQ content in the commercial BSO products ranged from 0.07% wt/wt to 1.88% wt/wt. The product with the highest TQ concentration is approximately 27-fold higher than the product with the lowest TQ concentration. Consequently, theoretical BSO amounts needed for specific diseases varied and some products cannot provide practical amount of TQ. This study recommends the regulation of TQ content in BSO and suggests that the BSO might be fortified with extra TQ to be effectively used in some diseases.

Matched MeSH terms: Chromatography, High Pressure Liquid
Troubleshooting and maintenance of high-performance liquid chromatography during herbicide analysis: an overview

Tayeb MA, Ismail BS, Khairiatul Mardiana J, Goh CT

Sains Malaysiana, 2016;45:237-245.

Glufosinate ammonium or ammonium salt (ammonium-(2RS)-2-amino-4- (methylphosphinato) butyric acid; C5H15N2O4P) is a commonly used polar herbicide in Malaysia and present in a variety of environmental waters at the sub-ppb level. Thus, glufosinate ammonium is analyzed in soil and water using high-performance liquid chromatography (HPLC), which is a complex yet the most powerful analysis tool. HPLC is tremendously sensitive and highly automated and HPLC instrumentation and machinery have improved over the years. However, typical problems are still encountered. HPLC users and advanced learners require help in identifying, separating and correcting typical problems. All HPLC systems consist of similar basic components. Although it is a modular system, trouble can occur in each component and change the overall performance. Resolving these problems may be expensive. This review describes the different aspects of HPLC, particularly troubleshooting, common problems and easy guidelines for maintenance.

Matched MeSH terms: Chromatography, High Pressure Liquid
Fulltext LC-MS data for metabolomics analysis of Garcinia mangostana L. seed germination

Mazlan O, Aizat WM, Aziz Zuddin NS, Baharum SN, Noor NM

Data Brief, 2018 Dec;21:2221-2223.
PMID: 30555858 DOI: 10.1016/j.dib.2018.11.072

Metabolic regulation is important during seed germination for the establishment of seedling. The germination strategy of mangosteen (Garcinia mangostana L.) seed is thought to be unique due to its recalcitrant characteristic (sensitive to coldness and drying). To investigate the metabolic changes during seed germination, we performed metabolomics analysis on germinating mangosteen seed sown after zero, one, three, five, seven and nine days. Sampled mangosteen seeds were subjected to methanol extraction prior analysis using Liquid Chromatography-Time of Flight-Mass Spectrometry (LC-TOF-MS). MS data were further analyzed using ProfileAnalysis (version 2.1). This is one of the earliest reports in metabolite identification and profiling of mangosteen seed at different germination stages. This data article refers to the article entitled "Metabolite profiling of mangosteen seed germination highlights metabolic changes related to carbon utilization and seed protection" (Mazlan et al., 2019) [1].

Matched MeSH terms: Chromatography, Liquid
Dataset on LC-Q-TOF/MS tentative identification of phytochemicals in the extract of Vernonia amygdalina leaf through positive ionization

Alara OR, Abdurahman NH, Ukaegbu CI, Hassan Z, Kabbashi NA

Data Brief, 2018 Dec;21:1686-1689.
PMID: 30505901 DOI: 10.1016/j.dib.2018.10.159

The tentative identification of bioactive compounds in the extract of Vernonia amygdalina leaf was carried out using positive ionization of Liquid chromatography-mass spectrometry quadrupole time of flight (LC-Q-TOF/MS). The positive ionization is associated with the presence of saponins, flavonoids, alkaloids, terpenoids, and glycosides. Tentative assignments of the secondary metabolites were performed by comparing the MS fragmentation patterns with Waters® UNIFY library which allows positive identification of the compounds based on the spectral match. All the metabolites compounds were estimated and presented in a BPI (Base peak intensity) plot. These data are the unpublished supplementary materials related to "Ethanolic extraction of bioactive compounds from V. amygdalina leaf using response surface methodology as an optimization tool" (Alara et al., 2018).

Matched MeSH terms: Chromatography, Liquid
Fulltext Chemical profile of Xanthine Oxidase inhibitor fraction of Persicaria Hydropiper

Noor Hashim, N.H., Maulidiani, M., Mediani, A., Abas, F.

International Food Research Journal, 2018;25(3):1088-1094.
MyJurnal

Persicaria hydropiper, locally known as kesum, is an herb belongs to the family Polygonaceae. It has been used widely in many countries as food flavoring and possesses a wide range of medicinal values. The total phenolic content and xanthine oxidase inhibitory activity of the methanolic extract of P. hydropiper and fractions were determined spectrophotometrically. The butanol fraction was found to contain high phenolic content and was able to inhibit xanthine oxidase activity. Online profiling using liquid chromatography coupled with electrospray ionisation spectrometry (LC-ESIMS/MS) has revealed ten constituents in this active fraction. The major components were flavonoid derivatives and flavonoid sulphates, which were confirmed by comparison with an authentic standards as well as their MS/MS fragmentation patterns and UV spectra.

Matched MeSH terms: Chromatography, Liquid
Fulltext Data on the optimisation and validation of a liquid chromatography-high-resolution mass spectrometry (LC-HRMS) to establish the presence of phosphodiesterase 5 (PDE5) inhibitors in instant coffee premixes

Mohd Yusop AY, Xiao L, Fu S

Data Brief, 2019 Aug;25:104234.
PMID: 31384643 DOI: 10.1016/j.dib.2019.104234

This paper presents the data on the optimisation and validation of a liquid chromatography-high-resolution mass spectrometry (LC-HRMS) to establish the presence of phosphodiesterase 5 (PDE5) inhibitors and their analogues as adulterants in instant coffee premixes. The method development data covered chromatographic optimisation for better analyte separation and isomeric resolution, mass spectrometry optimisation for high sensitivity and sample preparation optimisation for high extraction recovery (RE) and low matrix effect (ME). The validation data covered specificity, linearity, range, accuracy, limit of detection, limit of quantification, precisions, ME, and RE. The optimisation and validation data presented here is related to the article: "Determination of phosphodiesterase 5 (PDE5) inhibitors in instant coffee premixes using liquid chromatography-high-resolution mass spectrometry (LC-HRMS)" Mohd Yusop et al., 2019.

Matched MeSH terms: Chromatography, Liquid
Highly sensitive and selective determination of malathion in vegetable extracts by an electrochemical sensor based on Cu-metal organic framework

Al'Abri AM, Abdul Halim SN, Abu Bakar NK, Saharin SM, Sherino B, Rashidi Nodeh H, et al.

J Environ Sci Health B, 2019;54(12):930-941.
PMID: 31407615 DOI: 10.1080/03601234.2019.1652072

This article demonstrates the first application of a copper-based porous coordination polymer (BTCA-P-Cu-CP) as a carbon paste electrode (CPE) modifier for the detection of malathion. The electrochemical behavior of BTCA-P-Cu-CP/CPE was explored using cyclic voltammetry (CV) while chrono-amperometry methods were applied for the analytical evaluation of the sensor performance. Under optimized conditions, the developed sensor exhibited high reproducibility, stability, and wide dynamic range (0.6-24 nM) with the limits of detection and sensitivity equal to 0.17 nM and 5.7 µAnMcm-1, respectively, based on inhibition signal measurement. Furthermore, the presence of common coexisting interfering species showed a minor change in signals (<4.4%). The developed sensor has been applied in the determination of malathion in spiked vegetable extracts. It exhibited promising results in term of fast and sensitive determination of malathion in real samples at trace level with recoveries of 91.0 to 104.4%. (RSDs < 5%, n = 3). A comparison of the two studied techniques showed that the HPLC technique is unable to detect malathion when the concentration is lower than 1.8 µM while 0.006 µM is detected with appropriate RSDs 0.2-5.2% (n = 3) by amperometric method. Due to the high sensitivity and selectivity, this new electrochemical sensor will be useful for monitoring trace malathion in real samples.

Matched MeSH terms: Chromatography, High Pressure Liquid
Fulltext Biogenic amine content in various types of tofu: occurrence, validation and quantification

Yue, C.S., Ng, Q.N., Lim, A.K., Lam, M. H., Chee, K.N.

International Food Research Journal, 2019;26(3):999-1009.
MyJurnal

In the present work, the biogenic amines tryptamine (TRP), putrescine (PUT), histamine (HIS), tyramine (TYR) and spermidine (SPD) were determined in 32 various types of tofu that were obtained from different states in Malaysia. Three main types of tofu; soft tofu, firm tofu and processed tofu, were analysed in the present work. The biogenic amine contents in the respective types of tofu were analysed by a reversed-phase HPLC with a DAD detector after the aqueous extraction and derivatisation with dansyl chloride. The LOD values ranged from 0.019 mg/L for PUT to 0.028 mg/L for TYR. While, the LOQ values ranged from 0.063 mg/L (PUT) to 0.096 mg/L (TYR). The recovery values for all the five amines ranged from 80.3% to 120.5% with RSD ≤ 3.1%. The total levels of biogenic amines found varied, ranging from 1.5 mg/kg to 687.9 mg/kg, with mean values (p < 0.05) in descending order of 44.6, 12.6, 9.1, 4.8 and 4.7 mg/kg for PUT, TYR, SPD, HIS and TRP, respectively. PUT and TRP were the most prevailing biogenic amines and they were found respectively in 90.62% of the tofu analysed. Significant positive correlations (r = 0.266 to 0.874, p < 0.05) were found between some individual biogenic amines and protein content in all the three types of tofu. However, negative correlations (r = -0.246 to -0.832, p < 0.05) were observed between biogenic amines and moisture content, and between biogenic amines and water activity in all the three types of tofu. Significant and strong correlations (r = 0.525 to 0.999, p < 0.05) were found between most of the individual biogenic amines and the total biogenic amines. Those tofu exceeding the legal limits may affect the health of sensitive individuals.

Matched MeSH terms: Chromatography, High Pressure Liquid
Comparative secondary metabolites profiling and biological activities of aerial, stem and root parts of Salvadora oleoides decne (Salvadoraceae)

Saleem H, Ahmad I, Zengin G, Mahomoodally FM, Rehman Khan KU, Ahsan HM, et al.

Nat Prod Res, 2020 Dec;34(23):3373-3377.
PMID: 30678488 DOI: 10.1080/14786419.2018.1564299

In this study, different parts (aerial, stem and root) of Salvadora oleoides Decne were investigated in order to explore their phytochemical composition and biological potential. The bioactive contents were evaluated by conventional spectrophotometric methods. Additionally, the secondary metabolite compounds were identified by UHPLC-MS analysis. Biological potential was evaluated by determining antioxidant (DPPH, FRAP, and Phosphomolybdenum) and enzyme inhibitory (butrylcholinesterase and lipoxygenase) effects. Higher total bioactive contents were found in methanolic extracts which tend to correlate with higher radical scavenging and reducing potential of these extracts. LC/MS spectrum revealed the presence of 16 different secondary metabolites belonging to terpene, glucoside and sesquiterpenoid dervivatives. Glucocleomin and emotin A were the main compounds present in all three parts. The strongest butrylcholinesterase and lipoxygenase inhibitory activity was observed for root and stem DCM extracts. Demonstrated biological potential of S. oleoides plant can trace a new road map for developing newly designed bioactive pharmaceuticals.

Matched MeSH terms: Chromatography, High Pressure Liquid
Fulltext Isolation and partial characterization of Asian sea bass (Lates calcarifer) Vitellogenin

Fazielawanie NM, Siraj SS, Harmin SA, Ina-Salwany MY

Fish Physiol Biochem, 2013 Apr;39(2):191-200.
PMID: 22878544 DOI: 10.1007/s10695-012-9690-5

A study was conducted to isolate, partial characterize Asian sea bass (Lates calcarifer) vitellogenin (vtg). Two-year-old juvenile L. calcarifer (n = 10) were given three intraperitoneal injections of 17-β estradiol (E2) at a dose of 2 mg/kg body weight to induce vitellogenesis. Blood was collected 3 days after the last injection, and plasma was purified through gel filtration chromatography. A broad single symmetrical peak consisting of vtg molecule was produced. Protein concentration was 0.059 mg/ml as determined by Bradfrod assay using bovine serum albumin as a standard. The protein appeared as one circulating form in Native PAGE considering the dimeric form of putative vtg with molecular weight of 545 kDa. In SDS-PAGE under reducing conditions, two major bands appeared at 232.86 and 118.80 kDa and minor bands at 100.60, 85.80 and 39.92 kDa, respectively. The purified vtg was used to generate a polyclonal antibody, and the specificity of antibody was assessed by Western blot analysis. Two major bands were immunoreacted, but no cross-reactivity was observed with plasma from non-induced males. The protein was characterized as phosphoglycolipoprotein as it positively stained for the presence of lipid, phosphorus and carbohydrate using Sudan Black B, methyl green and periodic acid/Schiff reagent solution, respectively. The amino acid composition was analyzed by high sensitivity amino acid analysis that showed high percentage of non-polar amino acids (~48 %). The results suggest the potential utilization of vtg as a basis tool to further study about reproductive physiology of this important economical species.

Matched MeSH terms: Chromatography, Gel/veterinary; Chromatography, High Pressure Liquid/veterinary
Fulltext Proteomic Analysis of the Human Anterior Pituitary Gland

Yelamanchi SD, Tyagi A, Mohanty V, Dutta P, Korbonits M, Chavan S, et al.

OMICS, 2018 12;22(12):759-769.
PMID: 30571610 DOI: 10.1089/omi.2018.0160

The pituitary function is regulated by a complex system involving the hypothalamus and biological networks within the pituitary. Although the hormones secreted from the pituitary have been well studied, comprehensive analyses of the pituitary proteome are limited. Pituitary proteomics is a field of postgenomic research that is crucial to understand human health and pituitary diseases. In this context, we report here a systematic proteomic profiling of human anterior pituitary gland (adenohypophysis) using high-resolution Fourier transform mass spectrometry. A total of 2164 proteins were identified in this study, of which 105 proteins were identified for the first time compared with high-throughput proteomic-based studies from human pituitary glands. In addition, we identified 480 proteins with secretory potential and 187 N-terminally acetylated proteins. These are the first region-specific data that could serve as a vital resource for further investigations on the physiological role of the human anterior pituitary glands and the proteins secreted by them. We anticipate that the identification of previously unknown proteins in the present study will accelerate biomedical research to decipher their role in functioning of the human anterior pituitary gland and associated human diseases.

Matched MeSH terms: Chromatography, Liquid

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