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Investigation of interaction studies of cefpirome with ACE-inhibitors in various buffers

Nawaz M, Arayne MS, Sultana N, Abbas HF

Spectrochim Acta A Mol Biomol Spectrosc, 2015 Feb 25;137:1050-4.
PMID: 25300038 DOI: 10.1016/j.saa.2014.08.152

This work describes a RP-HPLC method for the determination and interaction studies of cefpirome with ACE-inhibitors (captopril, enalapril and lisinopril) in various buffers. The separation and interaction of cefpirome with ACE-inhibitors was achieved on a Purospher Star, C18 (5 μm, 250×4.6 mm) column. Mobile phase consisted of methanol: water (80:20, v/v, pH 3.3); however, for the separation of lisinopril, it was modified to methanol-water (40:60, v/v, pH 3.3) and pumped at a flow rate of 1 mL min(-1). In all cases, UV detection was performed at 225 nm. Interactions were carried out in physiological pH i.e., pH 1 (simulated gastric juice), 4 (simulated full stomach), 7.4 (blood pH) and 9 (simulated GI), drug contents were analyzed by reverse phase high performance liquid chromatography. Method was found linear in the concentration range of 1.0-50.0 μg mL(-1) with correlation coefficient (r(2)) of 0.999. Precision (RSD%) was less than 2.0%, indicating good precision of the method and accuracy was 98.0-100.0%. Furthermore, cefpirome-ACE-inhibitors' complexes were also synthesized and results were elucidated on the basis of FT-IR, and (1)H NMR. The interaction results show that these interactions are pH dependent and for the co-administration of cefpirome and ACE-inhibitors, a proper interval should be given.

Matched MeSH terms: Chromatography, High Pressure Liquid/methods; Chromatography, Reverse-Phase/methods
Validation of a reverse-phase high-performance liquid chromatography method for the determination of amino acids in gelatins by application of 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate reagent

Azilawati MI, Hashim DM, Jamilah B, Amin I

J Chromatogr A, 2014 Aug 1;1353:49-56.
PMID: 24797394 DOI: 10.1016/j.chroma.2014.04.050

In-house method validation was conducted to determine amino acid composition in gelatin by a pre-column derivatization procedure with the 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate reagent. The analytical parameters revealed that the validated method was capable of selectively performing a good chromatographic separation for 18 amino acids in less than 40 min; the overall detection and quantitation limit for amino acids fell into ranges of 5.68-12.48 and 36.0-39.0 pmol/μl, respectively; the matrix effect was not observed, and the linearity range was 37.5-1000 pmol/μl. The accuracy (precision and recovery) analyses of the method were conducted under repeatable conditions on different days in random order. Method precision revealed by HorRat values was significantly less than 2, except for histidine with a precision of 2.19, and the method recoveries had a range of 80-115% except for alanine which was recovered at 79.4%. The findings were reproducible and accurately defined, and the method was found to be suited to routine analysis of amino acid composition in gelatin-based ingredients.

Matched MeSH terms: Chromatography, High Pressure Liquid/methods*; Chromatography, Reverse-Phase/methods*
Application of supercritical fluid chromatography in the quantitative analysis of minor components (carotenes, vitamin E, sterols, and squalene) from palm oil

Choo YM, Ng MH, Ma AN, Chuah CH, Hashim MA

Lipids, 2005 Apr;40(4):429-32.
PMID: 16028723

The application of supercritical fluid chromatography (SFC) coupled with a UV variable-wavelength detector to isolate the minor components (carotenes, vitamin E, sterols, and squalene) in crude palm oil (CPO) and the residual oil from palm-pressed fiber is reported. SFC is a good technique for the isolation and analysis of these compounds from the sources mentioned. The carotenes, vitamin E, sterols, and squalene were isolated in less than 20 min. The individual vitamin E isomers present in palm oil were also isolated into their respective components, alpha-tocopherol, alpha-tocotrienol, gamma-tocopherol, gamma-tocotrienol, and delta-tocotrienol. Calibration of all the minor components of palm as well as the individual components of palm vitamin E was carried out and was found to be comparable to those analyzed by other established analytical methods.

Matched MeSH terms: Chromatography, Gas; Chromatography, High Pressure Liquid; Chromatography, Supercritical Fluid*
A new oxolane from Enterobacter cloacae

Yap AC, Chan KG, Sim KS, Choo YM

Nat Prod Res, 2016 Apr;30(7):783-8.
PMID: 26252083 DOI: 10.1080/14786419.2015.1065492

Enterobacter cloacae is a highly pathogenic Gram-negative proteobacterium which is responsible for a wide array of infections. In the present study, the fermentation culture of E. cloacae has yielded one new oxolane compound, Rimboxo (1) in addition to three known compounds, i.e. Maculosine (2), phenylacetic acid (3) and methyl myristate (4). These compounds were isolated and characterised using extensive chromatographic and spectroscopic methods, and were subjected to cytotoxicity evaluations.

Matched MeSH terms: Chromatography
Assay of 3-hydroxy-3-methylglutaryl CoA reductase activity using anionic-exchange column chromatography

Ong KK, Khor HT, Tan DT

Anal Biochem, 1991 Aug 01;196(2):211-4.
PMID: 1776669

A rapid, easy, and sensitive method is described in this paper for the assay of 3-hydroxy-3-methylglutaryl CoA (HMG CoA) reductase, a key enzyme in cholesterol biosynthesis. [14C]HMG CoA was used as the substrate and the product formed, i.e., [14C]mevalonate, was allowed to be converted to its lactone form (mevalonolactone) in the presence of HCl. The reaction mixture was applied to a column containing an anionic exchanger. The column was made up of QAE-Sephadex (A25, formate form) packed to a height of 4 cm in Pasteur pipets. Under these conditions, mevalonolactone was not retained by the column and was eluted with ammonium formate solution while HMG CoA, being negatively charged, was retained by the gel and eluted by HCl above 0.05 M. Determination of the amount of radioactivity in mevalonolactone was then used to quantitate the activity of HMG CoA reductase. This assay has been successfully used for determining the activity of this enzyme in a microsomal fraction prepared from the liver of the rat.

Matched MeSH terms: Chromatography, Ion Exchange/methods*; Chromatography, Thin Layer/methods
Complete fatty acid analysis data of flaxseed oil using GC-FID method

Danish M, Nizami M

Data Brief, 2019 Apr;23:103845.
PMID: 31372470 DOI: 10.1016/j.dib.2019.103845

The data presented in this article were generated through the gas chromatography (GC) with a flame ionization detector (FID). The flaxseed oil was converted into fatty acid methyl ester (FAME) then used in the GC with FID and observe the retention time of different fatty acid present in the flaxseed oil. The observed retention time was compared with the standard fatty acid to confirm the specific fatty acid presence in the flaxseed oil. The part of the data is used in the article "Optimization of the process variable for biodiesel production by transesterification of flaxseed oil and produced biodiesel characterizations" Renewable Energy journal (Ahmad et al., 2019).

Matched MeSH terms: Chromatography, Gas
Evaluation of a method to determine the natural occurrence of aflatoxins in commercial traditional herbal medicines from Malaysia and Indonesia

Ali N, Hashim NH, Saad B, Safan K, Nakajima M, Yoshizawa T

Food Chem Toxicol, 2005 Dec;43(12):1763-72.
PMID: 16019122

Traditional herbal medicines, popularly known as 'jamu' and 'makjun' in Malaysia and Indonesia, are consumed regularly to promote health. In consideration of their frequent and prolonged consumption, the natural occurrence of aflatoxins (AF) in these products was determined using immunoaffinity column clean-up and high-performance liquid chromatography with pre-column derivatization. The evaluated method, which entails dilution of sample extracts with Tween 20-phosphate buffered saline (1:9, v/v) and a chromatographic system using isocratic mobile phase composed of water-methanol-acetonitrile (70:20:10, v/v/v), was effective in separating AFB1, AFG1 and AFG2 from interference at their retention times. Results were confirmed using post-column derivatization with photochemical reactor. For 23 commercial samples analyzed, mean levels (incidence) of AFB(1), AFB(2) and AFG1 in positive samples were 0.26 (70%), 0.07 (61%) and 0.10 (30%) microg/kg, respectively; one sample was positive for AFG2 at a level of 0.03 (4%) microg/kg. In contrast to the high levels of AF in crude herbal drugs and medicinal plants reported previously by other researchers, the low contamination levels reported in this study may be attributed to the higher selectivity to AF of the method applied. Based on the AFB1 levels and the daily consumption of positive samples, a mean probable daily intake of 0.022 ng/kg body weight was calculated.

Matched MeSH terms: Chromatography, Affinity/methods; Chromatography, High Pressure Liquid/methods
Polycyclic Aromatic Hydrocarbons: Occurrence, Electroanalysis, Challenges, and Future Outlooks

Zainal PNS, Alang Ahmad SA, Abdul Aziz SFN, Rosly NZ

Crit Rev Anal Chem, 2020 Nov 06.
PMID: 33155481 DOI: 10.1080/10408347.2020.1839736

The past several decades have seen increasing concern regarding the wide distribution of polycyclic aromatic hydrocarbons (PAHs) in environmental matrices. Primary toxicological data show PAHs' persistent characteristics and possible toxicity effects. Because of this pressing global issue, electroanalytical methods have been introduced. These methods are effective for PAH determination in environmental waters, even outclassing sophisticated analytical techniques such as chromatography, conventional spectrophotometry, fluorescence, and capillary electrophoresis. Herein, the literature published on PAHs is reviewed and discussed with special regard to PAH occurrence. Moreover, the recent developments in electrochemical sensors for PAH determination and the challenges and future outlooks in this field, are also presented.

Matched MeSH terms: Chromatography
Extraction and identification of biologically important proteins from the medicinal plant God's crown (Phaleria macrocarpa)

Habib MAH, Ismail MN

J Food Biochem, 2021 07;45(7):e13817.
PMID: 34137461 DOI: 10.1111/jfbc.13817

The fruit and leaf of God's crown (Phaleria macrocarpa) have been traditionally used to treat a wide variety of diseases. However, the proteins of this tropical plant are still heavily understudied. Three protein extraction methods; phenol (Phe), trichloroacetic acid (TCA)-acetone-phenol (TCA-A-Phe), and ultrasonic (Ult) were compared on the fruit and leaf of P. macrocarpa. The Phe extraction method showed the highest percentage of recovered protein after the resolubilization process for both leaf (12.24%) and fruit (30.41%) based on protein yields of the leaf (6.15 mg/g) and fruit (36.98 mg/g). Phe and TCA-A-Phe extraction methods gave well-resolved bands over a wide range of molecular weights through sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Following liquid chromatography-tandem mass spectrometry analysis, proteins identified through the Phe extraction method were 30%-35% enzymatic proteins, including oxidoreductases, transferases, hydrolases, lyases, isomerases, and ligases that possess various biological functions. PRACTICAL APPLICATIONS: Every part of God's crown plant is traditionally consumed to treat various illnesses. While plant's benefits are well known and have led to a plethora of health products, the proteome remains mostly unknown. This study compares three protein extraction methods for the leaf and fruit of P. macrocarpa and identifies their proteins thru LC-MS/MS coupled with PEAKS. These method comparisons can be a guide for works on other plants as well. In addition, the proteomics data from this study may shed light on the functional properties of these plant parts and their products.

Matched MeSH terms: Chromatography, Liquid
Fulltext Caffeine and theobromine levels in chocolate couverture and coating products

Ramli N, Abdul Rahman S, Hassan O, Mohd Yatim A, Said M, Lim LS, et al.

Malays J Nutr, 2000 Mar;6(1):55-63.
PMID: 22692392

Thirty-two samples of chocolate products were analysed by HPLC for caffeine and theobromine contents. Defatted residues of samples were extracted with 80% aqueous acetone. After extraction into boiling water, the methylxanthines were identified and quantified with the use of μ-Bondapak column and mobile phase of methanol:water:acetic acid (20:79:1). Levels of caffein and theobromine in 32 samples of chocolate products averaged 0.62-1.14 mg/g and 0.026-0.153 mg/g respectively. Mean values for theobromine and caffeine content for chocolate coating were 0.82 and 0.07 mg/g respectively. The chocolate coating made from fat substitute had theobromine and caffeine levels ranging from 0.36-0.70 mg/g and 0.027-0.061 mg/g respectively, with mean values of 0.49 mg theobromine/g and 0.039 mg caffeine/g. In local chocolate, the mean theobromine and caffeine levels respectively were 0.72 mg/g and 0.04 mg/g in milk chocolate, and 0.85 mg/g and 0.06 mg/g in dark chocolate. Meanwhile, for imported chocolate, the mean theobromine and caffeine levels respectively were 1.05 mg/g and 0.12 mg/g in dark chocolate; 0.76 mg/g and 0.04 mg/g in milk chocolate; and 0.74 mg/g and 0.03 mg/g in white chocolate. Compared with the local chocolates, imported chocolates had higher levels of theobromine and caffeine at 1.141 mg/g and 0.1533mg/g. The average theobromine and caffeine concentrations in local chocolate were 0.082mg/g and 0.066mg/g. Theobromine concentration in chocolate samples is within the range of 0.62mg/g-1.141mg/g and the range of caffeine concentration is 0.026mg/g-0.153mg/g respectively. Bittersweet chocolates were found to have higher theobromine and caffeine concentrations than normal sweet chocolates and milk chocolates.

Matched MeSH terms: Chromatography, High Pressure Liquid
Fulltext Supercritical fluid extraction of bioactive flavonoid from Strobilanthes crispus (pecah kaca) and its comparison with solvent extraction

Liza, M.S., Abdul Rahman, R., Mandana, B., Jinap, S., Rahmat, A., Zaidul, I.S.M., et al.

International Food Research Journal, 2012;19(2):503-508.
MyJurnal

Supercritical carbon dioxide extraction (SC-CO2) of bioactive flavonoid from Strobilanthes crispus (Pecah Kaca) was performed to study the effects of various parameters such as pressure, temperature and dynamic extraction time on the yield and composition of bioactive flavonoid. The results were also compared with those obtained by conventional Soxhlet extraction in lab conditions. The results from SFE showed that the effect of extraction variables on extraction yields decreased in the following order: pressure, temperature and dynamic extraction time. The extraction pressure played a dominant role in the yield of the sample while the effect of time could be ignored. This study also revealed that both Soxhlet extraction and SC-CO2 extraction can be used to obtain flavonoid compound. Under the optimum conditions, the highest bioactive flavonoid compound content was obtained at 3.98% and eight flavonoid compounds were identified by HPLC.

Matched MeSH terms: Chromatography, High Pressure Liquid
Improvement of COD removal by controlling the substrate degradability during the anaerobic digestion of recalcitrant wastewater

Kawai M, Nagao N, Kawasaki N, Imai A, Toda T

J Environ Manage, 2016 Oct 01;181:838-846.
PMID: 27449962 DOI: 10.1016/j.jenvman.2016.06.057

The recalcitrant landfill leachate was anaerobically digested at various mixing ratios with labile synthetic wastewater to evaluate the degradation properties of recalcitrant wastewater. The proportion of leachate to the digestion system was increased in three equal steps, starting from 0% to 100%, and later decreased back to 0% with the same steps. The chemical oxygen demand (COD) for organic carbon and other components were calculated by analyzing the COD and dissolved organic carbon (DOC), and the removal efficiencies of COD carbon and COD others were evaluated separately. The degradation properties of COD carbon and COD others shifted owing to changing of substrate degradability, and the removal efficiencies of COD carbon and COD others were improved after supplying 100% recalcitrant wastewater. The UV absorptive property and total organic carbon (TOC) of each molecular size using high performance liquid chromatography (HPLC)-size exclusion chromatography (SEC) with UVA and TOC detectors were also investigated, and the degradability of different molecular sizes was determined. Although the SEC system detected extracellular polymeric substances (EPS), which are produced by microbes in stressful environments, during early stages of the experiment, EPS were not detected after feeding 100% recalcitrant wastewater. These results suggest that the microbes had acclimatized to the recalcitrant wastewater degradation. The high removal rates of both COD carbon and COD others were sustained when the proportion of labile wastewater in the substrate was 33%, indicating that the effective removal of recalcitrant COD might be controlled by changing the substrate's degradability.

Matched MeSH terms: Chromatography, Gel/methods; Chromatography, High Pressure Liquid/methods
Chemometric techniques in oil classification from oil spill fingerprinting

Ismail A, Toriman ME, Juahir H, Kassim AM, Zain SM, Ahmad WKW, et al.

Mar Pollut Bull, 2016 Oct 15;111(1-2):339-346.
PMID: 27397593 DOI: 10.1016/j.marpolbul.2016.06.089

Extended use of GC-FID and GC-MS in oil spill fingerprinting and matching is significantly important for oil classification from the oil spill sources collected from various areas of Peninsular Malaysia and Sabah (East Malaysia). Oil spill fingerprinting from GC-FID and GC-MS coupled with chemometric techniques (discriminant analysis and principal component analysis) is used as a diagnostic tool to classify the types of oil polluting the water. Clustering and discrimination of oil spill compounds in the water from the actual site of oil spill events are divided into four groups viz. diesel, Heavy Fuel Oil (HFO), Mixture Oil containing Light Fuel Oil (MOLFO) and Waste Oil (WO) according to the similarity of their intrinsic chemical properties. Principal component analysis (PCA) demonstrates that diesel, HFO, MOLFO and WO are types of oil or oil products from complex oil mixtures with a total variance of 85.34% and are identified with various anthropogenic activities related to either intentional releasing of oil or accidental discharge of oil into the environment. Our results show that the use of chemometric techniques is significant in providing independent validation for classifying the types of spilled oil in the investigation of oil spill pollution in Malaysia. This, in consequence would result in cost and time saving in identification of the oil spill sources.

Matched MeSH terms: Chromatography, Gas/methods; Gas Chromatography-Mass Spectrometry/methods*
Recent Modifications and Validation of QuEChERS-dSPE Coupled to LC-MS and GC-MS Instruments for Determination of Pesticide/Agrochemical Residues in Fruits and Vegetables: Review

Lawal A, Wong RCS, Tan GH, Abdulra'uf LB, Alsharif AMA

J Chromatogr Sci, 2018 Aug 01;56(7):656-669.
PMID: 29688338 DOI: 10.1093/chromsci/bmy032

Fruits and vegetables constitute a major type of food consumed daily apart from whole grains. Unfortunately, the residual deposits of pesticides in these products are becoming a major health concern for human consumption. Consequently, the outcome of the long-term accumulation of pesticide residues has posed many health issues to both humans and animals in the environment. However, the residues have previously been determined using conventionally known techniques, which include liquid-liquid extraction, solid-phase extraction (SPE) and the recently used liquid-phase microextraction techniques. Despite the positive technological effects of these methods, their limitations include; time-consuming, operational difficulty, use of toxic organic solvents, low selective property and expensive extraction setups, with shorter lifespan of instrumental performances. Thus, the potential and maximum use of these methods for pesticides residue determination has resulted in the urgent need for better techniques that will overcome the highlighted drawbacks. Alternatively, attention has been drawn recently towards the use of quick, easy, cheap, effective, rugged and safe technique (QuEChERS) coupled with dispersive solid-phase extraction (dSPE) to overcome the setback challenges experienced by the previous technologies. Conclusively, the reviewed QuEChERS-dSPE techniques and the recent cleanup modifications justifiably prove to be reliable for routine determination and monitoring the concentration levels of pesticide residues using advanced instruments such as high-performance liquid chromatography, liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry.

Matched MeSH terms: Chromatography, Liquid/methods*; Gas Chromatography-Mass Spectrometry/methods*
Fulltext Proteomic analysis of pitcher fluid from Nepenthes × ventrata

Wan Zakaria WNA, Aizat WM, Goh HH, Noor NM

Data Brief, 2018 Apr;17:517-519.
PMID: 29876422 DOI: 10.1016/j.dib.2018.01.037

The carnivorous plants of genus Nepenthes produce unique pitchers containing secretory glands, which secrete proteins into the digestive fluid. We investigated protein profile in the pitcher fluid during the first three days of opening to understand carnivory trait of Nepenthes × ventrata. The proteome analysis of pitcher fluid from N. × ventrata was performed by label-free quantitative liquid chromatography mass spectrometry (nLC-MS/MSALL). Raw MS data have been deposited to the ProteomeXchange with identifier PXD007251. This dataset allows the identification and quantification of proteins from pitcher fluids to elucidate proteins involved in carnivory physiology of Nepenthes species.

Matched MeSH terms: Chromatography, Liquid
Kesan kepekatan asid performik terhadap pengepoksidaan minyak sawit olein

Darfizzi Derawi, Jumat Salimon

Sains Malaysiana, 2011;40:1123-1127.

Kajian pengoptimuman tindak balas pengepoksidaan minyak sawit olein (POo) dengan menggunakan mangkin asid performik (HCOOOH) telah dijalankan. Kesan tindak balas terhadap nisbah kepekatan HCOOH : H2O2 dan masa tindak balas telah dikaji pada suhu 45oC. Kandungan oksigen oksirana (OOC) optimum sebanyak 3.61% diperoleh dengan menggunakan nisbah mol 1 : 5 : 2 pada 150 min dan sebanyak 96.5% penukaran dapat dicapai. Hasil tindak balas adalah sebanyak 91.3%. Puncak kromatografi cecair berprestasi tinggi (HPLC) bagi minyak sawit olein terepoksida (EPOo) telah berubah sepenuhnya berbanding puncak HPLC POo. Spektrum transformasi Fourier infra-merah (FTIR) telah menunjukkan kehadiran puncak ikatan gelang oksirana pada 844 cm-1.

Matched MeSH terms: Chromatography, High Pressure Liquid
Variasi dan sumber sterol di Kuala Selangor, Selangor

Masni Mohd Ali, Norfariza Humrawali, Mohd Talib Latif

Sains Malaysiana, 2010;39.

Kajian ini adalah mengenai peranan sterol sebagai penunjuk bio-lipid untuk mengenal pasti variasi dan sumber bahan organik yang hadir bersama sedimen permukaan dari Kuala Selangor, Selangor. Kajian melibatkan kaedah pengekstrakan sterol daripada sampel sedimen dan seterusnya dianalisis menggunakan GC-MS untuk menentukan kehadiran sebatian tersebut. Sepuluh sebatian sterol dikenal pasti hadir dengan fitosterol merupakan sebatian dominan di kawasan kajian iaitu 79% daripada jumlah keseluruhan sterol. Ini diikuti oleh kolesterol serta sterol kumbahan masing-masing menyumbangkan 6% daripada jumlah keseluruhan sterol manakala selebihnya berada dalam julat 1-5%. Indeks Sumber Sterol (SSI) juga menunjukkan kandungan fitosterol yang tinggi walaupun hadir pada kadar yang berbeza di setiap stesen pensampelan. Penilaian pencemaran kumbahan menggunakan nisbah koprostanol/kolesterol, koprostanol/(koprostanol+kholestanol) serta epikoprostanol/koprostanol menunjukkan kawasan kajian tidak mengalami pencemaran kumbahan walaupun sterol daripada sumber kumbahan hadir di persekitarannya. Kesimpulannya sedimen permukaan di Kuala Selangor, Selangor mengandungi campuran sterol daripada pelbagai sumber yang hadir di persekitarannya dengan didominasi oleh fitosterol yang berasal daripada tumbuhan terestrial.

Matched MeSH terms: Gas Chromatography-Mass Spectrometry
Interference of hemoglobinA1c (HbA1c) detection using ion-exchange high performance liquid chromatography (HPLC) method by clinically silent hemoglobin variant in University Malaya Medical Centre (UMMC)--a case report

Thevarajah M, Nadzimah MN, Chew YY

Clin Biochem, 2009 Mar;42(4-5):430-4.
PMID: 19026622 DOI: 10.1016/j.clinbiochem.2008.10.015

Glycated hemoglobin, measured as HbA1c is used as an index of mean glycemia in diabetic patients over the preceding 2-3 months. Various assay methods are used to measure HbA1c and many factors may interfere with its measurement according to assay method used, causing falsely high or low results.

Matched MeSH terms: Chromatography, High Pressure Liquid/methods*; Chromatography, Ion Exchange/methods*
Fulltext Simple and sensitive method for the analysis of ketorolac in human plasma using high-performance liquid chromatography

Muralidharan S, Jayaraja Kumar K, Parasuraman S

J Young Pharm, 2013 Sep;5(3):98-101.
PMID: 24396250 DOI: 10.1016/j.jyp.2013.06.007

To develop a simple and sensitive method of ketorolac in drug free human plasma using high-performance liquid chromatographic (HPLC).

Matched MeSH terms: Chromatography, High Pressure Liquid
Evaluation of role of HPLC (Merits & Pitfalls), in the diagnosis of various hemoglobinopathies & thalassemic syndromes

Baig MA, Swamy KB, Baksh AD, Bahashwan A, Moshrif Y, Al Sawat A, et al.

Indian J Pathol Microbiol, 2021 8 4;64(3):518-523.
PMID: 34341263 DOI: 10.4103/IJPM.IJPM_709_20

Background: : HPLC is one of the most important tools for accurate diagnosis of hemoglobinopathies and thalassemias. The advantage of the HPLC system is the excellent resolution, reproducibility &quantification of several normal and abnormal hemoglobin.
Results: BIO RAD Variant II analyzer was used. Sickle cell syndromes including double heterozygous states accounted for 56.13% of total cases. HbSS, HbS/β0-th, HbS/β+-th β-thal trait comprises 29%, 6.5%, 5.1%& 10% of total cases respectively with mean MCV (fl) = 84, 68,71,64 respectively. The Mean HbA2 for β-thal trait, HbE trait &HbE-β thal showed 5.1 ± 1.1, 19 ± 9 & 24 ± 8 respectively. HbF is increased in 8.6% case (excluding SC syndromes & β-thal disorders), of these 5.5% were infants & 12 cases of Aplastic Anemias. Peak P2 >7% (2.4% cases) was seen in uncontrolled diabetes mellitus which on quantification showed HbA1C = 8 ± 2.1 mmol/L.
Discussion: : HPLC in correlation with CBC parameters & family studies can aid in the diagnosis of majority of Hemoglobinopathies and thalassemic syndrome. The CBC & HPLC parameters of the present study are in good correlation with the research conducted by Tejinder Sing, RiouJ & Alla Joutovsky. Present study showed HPLC comprehensively characterizing HbS, A, A2, F, S, C, D from each other & was also applicable for the quantification of HbA1c for the monitoring of Diabetes Mellitus.
Conclusion: : The merits of HPLC are small quantity of sample required, economical, less TAT, accurate categorization of HbS, HbA2 & F. But one has to be aware of the limitations and problems associated with this method due to variant hemoglobin within the same retention windows. The present findings show HPLC as an excellent & powerful diagnostic tool for the direct identification of hemoglobin variants with a high degree of precision in the quantification of normal and abnormal hemoglobin fractions.

Matched MeSH terms: Chromatography, High Pressure Liquid/economics; Chromatography, High Pressure Liquid/methods*; Chromatography, High Pressure Liquid/standards*

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