Elution System Research Articles

Разработка способа экстракции бета-каротина для его определения в комплексных пищевых добавках

Introduction. Natural dye beta-carotene E160a (pro-vitamin A) is part of many functional foods. Therefore, the total intake of beta-carotene must not exceed the tolerable upper intake level, i.e. 10 mg/day. E160a is also used in formulations of complex food additives. These products contain antioxidants, emulsifiers, preservatives, carriers, stabilizers, and thickeners, which makes it make difficult to isolate and determine the amount of beta-carotene. Complex food additives vary in composition, which, together with incomplete extraction and degradation of beta-carotene, results in inaccuracy of its determination. The present research reviewed various techniques of beta-carotene extraction from liquid and dry complex food additives.
 Study objects and methods. The study featured commercial samples of complex food additives containing beta-carotene. It was isolated from the samples by liquid extraction methods using organic solvents and their binary mixtures. The extraction was performed in a stirring device at 120 rpm and in a Bandelin Sonorex ultrasonic bath at 128 W and 35 kHz. The extraction continued until the samples were completely discolored. The list of solvents included chloroform, petroleum ether, hexane, acetone, ethanol, and their mixes. The experiment was based on high performance thin layer chromatography with Sorbfil sheet plates. The optical density was determined by spectrophotometry using a SHIMADZU UV-1800 device.
 Results and discussion. The extraction of beta-carotene from liquid samples with mechanical stirring at 120 rpm led to the formation of stable opaque emulsions, which made it difficult to determine the amount of beta-carotene. The emulsions were destroyed by ultrasound exposure for 30 min. The degree of extraction of beta-carotene from liquid emulsion systems (≥ 95%) was achieved by ultrasonic extraction with chloroform or a binary extractant based on mixes of acetone with chloroform, hexane, or petroleum ether in volumetric ratios of 1:1. When extracted from dry samples using chloroform or binary extractants, beta-carotene was not isolated or was not completely isolated. As a result, pre-dispersion of dry samples in water with stirring at 120 rpm was proposed. Ultrasonic treatment did not contribute to the dispersion of dry samples due to insufficient cavitation action. Ethanol, which is a solubilizer, increased the extractability of beta-carotene up to 98 %. The optimal results were achieved by adding a binary extractant (ethanol:chloroform in a 1:2 ratio) to an aqueous solution of the dry sample (1:3). The optimal elution systems were hexane:benzene (9.7:0.3), butanol:methyl ethyl ketone:ammonia 10 % (9:6:6), and petroleum ether:benzene:acetone:acetic acid (23.3:5.8:0.6:0.3).
 Conclusion. The article offers new methods of beta-carotene isolation from liquid and dry complex food additives of various composition and elution systems for carotene identification. The methods caused no degradation of beta-carotene during extraction and can be used to prepare samples of complex food additives for beta-carotene identification and quantitative analysis.

Analysis of the Monosaccharide Composition of Water-Soluble Coriolus versicolor Polysaccharides by Ultra-Performance Liquid Chromatography/Photodiode Array Detector

Coriolus versicolor is a well-known edible and medicinal fungus containing polysaccharides and polysaccharopeptides, which have been proven to possess interesting bioactivities, especially antitumor and immunomodulatory activities. A facile and sensitive method for the monosaccharide composition analysis of polysaccharides in C. versicolor has been developed by employing precolumn derivatization ultra-performance liquid chromatography equipped with a photodiode array detector (UPLC-PDA). The hydrolysis conditions of polysaccharides from C. versicolor and the optimization of the sample precolumn derivatization with 1-phenyl-3-methyl-5-pyrazolone (PMP) were comprehensively studied. The LODs of the quantity of monosaccharides were in the range of 3.1–4.6 ng. The PMP-labeled mixture of monosaccharides could be successfully separated by UPLC and detected by a PDA detector under optimized conditions. The mobile phase of the elution system was chosen as acetonitrile and 25 mm K2HPO4 (pH 6.5) with the Phenomenex Kinetex C18 column, and the column temperature was set at 35 °C. The C. versicolor polysaccharides mainly consist of mannose (Man), glucose (Glc), galactose (Gal), fucose (Fuc), trace rhamnose (Rha), ribose (Rib), glucuronic acid (GlcA), galacturonic acid (GalA), xylose (Xyl), and arabinose (Ara), and the content of Glc is the highest.

Quantitative analysis of tryptophan and its metabolites in urine by ultra performance liquid chromatography-tandem mass spectrometry

基于超高效液相色谱-串联质谱(UPLC-MS/MS)建立定量分析色氨酸(Trp)及代谢产物3-OH-犬尿氨酸(3-OH-Kyn)、3-OH-邻氨基苯甲酸(3-OH-AA)、黄尿酸(XA)、犬尿氨酸(Kyn)、5-羟基吲哚乙酸(5-HIAA)、犬尿喹啉酸(KA)和5-羟色胺(5-HT)的方法,应用该方法分析其在尿样中的含量,探讨排泄规律。将尿样稀释、离心后,加入丹磺酰氯(DNS-Cl)衍生,经Thermo C18色谱柱(50 mm×3 mm, 2.7 μm)分离和0.1%甲酸和甲醇梯度洗脱后,采用电喷雾电离(ESI)源,在正离子扫描和多反应监测(MRM)模式下检测。以咖啡酸(CA)为内标,定量分析。结果显示,8种目标化合物的线性关系良好,相关系数(R 2)≥0.9740,检测灵敏(LOD为0.005~0.5 ng/mL),回收率高(93.24%~107.65%)。采用本方法检测分析了健康志愿者70个尿液样本,在尿样中检测到Trp原型及其7种代谢产物。结果表明,体内的Trp是通过原型和代谢两种方式排泄:Trp原型的含量为5.22~20.88 μg/mL;尿液中经代谢后排泄的Trp量是原型的124%~268%,即体内的Trp主要经代谢后排出体外。方法主要研究了Trp-5-HT和Trp-Kyn两条途径的代谢产物含量,Trp经Kyn降解生成的3-OH-AA和3-OH-Kyn含量较多,即Trp-Kyn是体内Trp的主要代谢途径。方法通过UPLC-MS/MS实现了尿液中Trp及其代谢产物含量的检测,能为临床检查提供技术和理论支持。

Simultaneous determination of saikosaponin derivatives in Bupleurum falcatum by HPLC-ELSD analysis using different extraction methods

Saikosaponin derivatives such as saikosaponins A, B1, B2, B3, B4, C, and D present in Bupleurum falcatum were analyzed by a high performance liquid chromatograph equipped with an evaporative light scattering detector, using different extraction solvents (water and 70% ethanol). The samples were injected into a YMC Pack Pro C18 column and separated using a gradient elution system with a mobile phase composed of acetonitrile and water at a flow rate of 1.1 mL/min. The content of saikosaponin derivatives was higher in 70% ethanol extract than in water extract. This study provides an efficient analytical method for determining the optimal conditions for extraction of saikosaponin derivatives, which can be used as a basis for development of functional foods and pharmaceutical products from B. falcatum.

Simultaneous determination of phytochemical constituents in Paeonia lactiflora extracts using the HPLC-UV method

Qantitative analysis of six compounds: (+)-catechin, benzoic acid, gallic acid methyl ester, paeonol, paeoniflorin, and albiflorin from Paeonia lactiflora extracts was performed using high-performance liquid chromatography and an ultraviolet (UV) detector, following different extraction methods. A reverse-phase column was used in a gradient elution system, and UV detection was performed at 280 nm. The results showed that the quantity of paeoniflorin was the highest in ethanol and water extracts (73.89 and 57.87 mg/g, respectively) among the six compounds. This study contributes a good analysis method for the contents of P. lactiflora and would be propitious for developing medicines and functional foods.

Exploiting column chemistry for chromatographic separation and quantification of caffeoylquinic acids in Gynura procumbens

AbstractA simple and sensitive high-performance liquid chromatography-ultraviolet (HPLC-UV) method was developed by exploiting the benefits of phenyl-hexyl column for the simultaneous determination of mono- and di-caffeoylquinic acids in Gynura procumbens plant samples. An optimal chromatographic separation was achieved by using a mobile phase of acetonitrile: 0.25% acetic acid in water (12.5:78.5, v/v) and detection at 330 nm. The limits of detection (LOD) and quantification (LOQ) for the six caffeoylquinic acid standards were in the range of 0.078–0.653 and 0.259–1.795 μg/mL, respectively. The accuracies of the developed method were in the range of 96.84–103.08%, while the corresponding precisions were between 0 and 2.94% for both within-day and between-day analyses, indicating that the method is repeatable and reliable. The mean recoveries were between 87.08 and 117.83%. The method was successfully applied for quantification of caffeoylquinic acids in G. procumbens plant samples. This is the first study on di-caffeoylquinic acids quantification in G. procumbens. Leaves samples contained higher amount of the caffeoylquinic acids compared to stem samples. Of the compounds, 3,5-dicaffeoylquinic acid was found to be the major compound in almost all G. procumbens samples. The method has advantages such as sensitive ultraviolet (UV) detection, short run time with simple isocratic elution system compared to other methods which involved the use of costly instruments, laborious procedures with long run time and complex gradient system. This method can be further extended for routine quality control and analysis of plants or herbal products containing the caffeoylquinic acids.

HPLC Method for Simultaneous Determination of Dextromethorphan Hydrobromide, Chlorpheniramine Maleate and Potassium Sorbate in Cough Syrup

The development and validation of an analytical method for the simultaneous determination of Dextromethorphan Hydrobromide, Chlorpheniramine Maleate, and Potassium Sorbate in cough syrup using the High-Performance Liquid Chromatography (HPLC) method have been conducted. The determination was carried out on the simulation sample consisting of three substances and prepared five different concentrations, with three replications for each concentration. The optimum HPLC conditions were obtained using the C18 column, with UV detection at 230 nm The mobile phase of acetonitrile & phosphate buffer pH 2.5 (containing sodium-heptane sulfonate as ion-pair), with a gradient elution system, at a 1 mL/minute, while the flow rate column temperature is set at 35oC. The results showed that the method has met the parameters set in the validation test with the recovery ranged from 98% - 102%, the precision test results (RSD) < 2%, and the linearity (r) ≥ 0.98. The method obtained is quite selective by confirmed identity with the maximum wavelength scanning of each analyte peak by PDA detector. The limit of detection (LoD) for Dextromethorphan-HBr, Chlorpheniramine maleate and Potassium sorbate were 3.13 µg/mL, 0.94 µg/mL and 0.12 µg/mL respectively and the limit of quantitation (LoQ) were 9.49 µg/mL, 2.85 µg/mL and 0.38 µg/mL respectively.

Identification and Characterization of Chemical Constituents in HuaTanJiangQi Capsules by UPLC-QTOF-MS Method.

HuaTanJiangQi (HTJQ) is a classical Chinese medicine compound preparation, mainly used for clinically treating and improving chronic obstructive pulmonary disease (COPD) in China. To establish a rapid and efficient analytical method for the identification and characterization of chemical constituents in HTJQ based on ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS). UPLC-QTOF-MS was used to rapidly separate and identify the chemical constituents of HTJQ via a gradient elution system. The accurate mass data of the protonated and deprotonated molecules and fragment ions were detected in positive and negative ion modes. Compounds of HTJQ can be identified and assigned by analyzing accurate mass measurements and ion fragmentation mechanisms and comparing them with a chemical compositions database. A total of 61 compounds in HTJQ were separated and identified, including 14 flavonoids, 16 organic acids, four isothiocyanic acids, eight butyl phthalides, two alkaloids, 10 terpenoids, four methoxyphenols and furanocoumarins, and three other compounds. The chemical compounds of HTJQ were identified and elucidated comprehensively for the first time. A rapid, accurate, and efficient UPLC-QTOF-MS method has been developed for the identification of chemical components and applied to simultaneously evaluate the quality and effectiveness of HTJQ. Systematic identification of chemical constituents in HTJQ can provide a scientific and reasonable basis for the application of HTJQ in the clinical treatment of COPD.

Biogenic Amines as Freshness Indicator in Halal and Non-Halal Slaughtered Chicken Meat Using Chromatographic Approach

Biogenic amines are naturally occurring organic bases produced by bacterial decarboxylation of amino acids that have been associated with harmful toxicology effects to humans. The main objective of this work is to investigate the possible role of biogenic amines (BAs) as indicators of spoilage in fresh chicken meat stored at 4°C for 15 days. A reversed-phase high-performance liquid chromatographic method with isocratic elution system is used for the quantification of four biogenic amines (putrescine, histamine, tyramine, cadaverine and spermine as well as spermidine) in chicken meat. Amines were extracted with 5% of trichloroacetic acid (TCA) and derivatised using dansyl chloride. The variation storage time differentiated the chicken meat on the microbiological characteristics. The results obtained shows that histamine, spermidine, tyramine, cadaverine, and putrescine increased slowly while spermine decreased for both chicken breast in halal and non-halal chicken meat. Thus, this BAs could be used as a spoilage index of fresh chicken meat.

Determination and Pharmacokinetics of Omeprazole Enantiomers in Human Plasma and Oral Fluid Utilizing Microextraction by Packed Sorbent and Liquid Chromatography-Tandem Mass Spectrometry.

In the present work, the determination of omeprazole (OME) enantiomers in oral fluid and plasma samples was carried out utilizing microextraction by packed sorbent (MEPS) and liquid chromatography-tandem mass spectrometry. A chiral column with cellulose-SB phase was used for the first time for enantiomeric separation of OME with an isocratic elution system using 0.2% ammonium hydroxide in hexane-ethanol mixture (70 : 30, v/v) as the mobile phase. OME enantiomers were determined utilizing a triple quadrupole tandem mass spectrometer in positive ion mode (ESI+) monitoring mass transitions: m/z 346.3 ⟶ 198.0 for OME and m/z 369.98 ⟶ 252.0 for internal standard. The limits of detection and quantification of the present method for both enantiomers were 0.1 and 0.4 ng/mL, respectively. The method validation provided good accuracy and precision. The matrix effect factor was less than 5%, and no interfering peaks were observed. The interday precision values ranged from 2.2 to 7.5 (%RSD), and the accuracy of determinations varied from −9.9% to 8.3%. In addition, the pharmacokinetics (PK) of omeprazole enantiomers in healthy subjects after a single oral dose was investigated. (S)-Enantiomers showed higher levels than (R)-enantiomers throughout 24 h. It was found that the mean maximum concentrations of (R)- and (S)-omeprazole in plasma samples were about two times higher than in oral fluid.

Development and Validation of a Method to Simultaneously Determine Multiple Sterols in Diversiform Food Substrates with UPLC-MS/MS

Sterols play an essential part in physiological activities of almost all creatures. However, there is a lack of a widely applicable routine method to investigate various sterols with both saturated and unsaturated forms in diversiform food materials. In this study, a new UPLC-MS/MS method with wider extensive applicability has been developed to fill this blank, which is considered time and cost saving, and can improve labor efficiency. The Agilent Eclipse Plus C18 column and methanol-water gradient elution system were employed to improve the separation at 40 °C and the flow rate was 0.5 mL/min. The method validation parameters include precision, recovery, linearity, and limit of detection and quantification (LOD and LOQ). The recoveries of sterols in all food matrices and the inter-day and intra-day precisions represented by RSDs were all within the recommended range of chemical analysis. The LODs and LOQs in different types of samples were in range of 0.0036–0.68 μg/mL and 0.01–2.0 μg/mL, respectively. All the results demonstrated that this method was sensitive and accurate enough. Additionally, the acid hydrolysis was found closely related to the degradation of unsaturated sterols in the pretreatment.

Biogenic Amines as Freshness Indicator in Halal and Non-Halal Slaughtered Chicken Meat Using Chromatographic Approach

Biogenic amines are naturally occurring organic bases produced by bacterial decarboxylation of amino acids that have been associated with harmful toxicology effects to humans. The main objective of this work is to investigate the possible role of biogenic amines (BAs) as indicators of spoilage in fresh chicken meat stored at 4°C for 15 days. A reversed-phase high-performance liquid chromatographic method with isocratic elution system is used for the quantification of four biogenic amines (putrescine, histamine, tyramine, cadaverine and spermine as well as spermidine) in chicken meat. Amines were extracted with 5% of trichloroacetic acid (TCA) and derivatised using dansyl chloride. The variation storage time differentiated the chicken meat on the microbiological characteristics. The results obtained shows that histamine, spermidine, tyramine, cadaverine, and putrescine increased slowly while spermine decreased for both chicken breast in halal and non-halal chicken meat. Thus, this BAs could be used as a spoilage index of fresh chicken meat.

Biophysical characterization of antibacterial compounds derived from pathogenic fungi Ganoderma boninense.

There have been relatively few studies which support a link between Ganoderma boninense, a phytopathogenic fungus that is particularly cytotoxic and pathogenic to plant tissues and roots, and antimicrobial compounds. We previously observed that liquid-liquid extraction (LLE) using chloroformmethanol-water at a ratio (1:1:1) was superior at detecting antibacterial activities and significant quantities of antibacterial compounds. Herein, we demonstrate that antibacterial secondary metabolites are produced from G. boninense mycelia. Antibacterial compounds were monitored in concurrent biochemical and biophysical experiments. The combined methods included high performance thin-layer chromatography (HPTLC), gas chromatography-mass spectrometry (GC-MS), high-performance liquid chromatography (HPLC), fourier transform infrared (FTIR), and nuclear magnetic resonance (NMR) spectroscopy. The antibacterial compounds derived from mycelia with chloroform-methanol extraction through LLE were isolated via a gradient solvent elution system using HPTLC. The antibacterial activity of the isolated compounds was observed to be the most potent against Staphylococcus aureus ATCC 25923 and multidrug-resistant S. aureus NCTC 11939. GC-MS, HPLC, and FTIR analysis confirmed two antibacterial compounds, which were identified as 4,4,14α-trimethylcholestane (m/z = 414.75; lanostane, C30H54) and ergosta-5,7,22-trien-3β-ol (m/z = 396.65; ergosterol, C28H44O). With the aid of spectroscopic evaluations, ganoboninketal (m/z = 498.66, C30H42O6), which belongs to the 3,4-seco-27-norlanostane triterpene family, was additionally characterized by 2D-NMR analysis. Despite the lack of antibacterial potential exhibited by lanostane; both ergosterol and ganoboninketal displayed significant antibacterial activities against bacterial pathogens. Results provide evidence for the existence of bioactive compounds in the mycelia of the relatively unexplored phytopathogenic G. boninense, together with a robust method for estimating the corresponding potent antibacterial secondary metabolites.Electronic Supplementary MaterialSupplementary material is available for this article at 10.1007/s12275-021-0551-8 and is accessible for authorized users.

Simultaneous determination of five bioactive components of XiaoJin Capsule in normal and mammary gland hyperplasia rat plasma using LC-MS/MS and its application to a pharmacokinetic study.

XiaoJin Capsule (XJC) is a classic Traditional Chinese Medicine formula for clinical treatment of thyroid nodules, mammary gland hyperplasia and breast cancer. For the specification and rational application of XJC in the future, an accurate and specific LC-MS/MS method was developed and validated for quantitative determination of five components in rat plasma after oral administration of XJC. The collected plasma samples were extracted by protein precipitation with methanol-acetonitrile (1:3, v/v) mixture solvent and separated on a C18 column using a gradient elution system. Mass spectrometry was performed on a triple quadrupole mass spectrometer, and samples were detected in positive ionization and multiple reactions monitoring mode. The method was properly validated in terms of linearity, precision, accuracy, recovery, matrix effect and stability. All calibration curves showed good linearity (r2 > 0.9910) over their concentration ranges. The intra- and inter-day precisions (RSD) were within 11.0%, and the LLOQ was 0.1, 0.2, 0.5, 7.5 and 7.5 ng/ml for aconine, songorine, neoline, 3-acetyl-11-keto-β-boswellic acid and 11-keto-β-boswellic acid, respectively. Extraction recovery, matrix effect and stability were satisfactory in rat plasma. This established method was successfully applied to a pharmacokinetics study of five compounds after oral administration of XJC to normal and mammary gland hyperplasia model rats.

A Simple and Rapid High-Performance Liquid Chromatography-Photodiode Array Method for Quantitation of Triacetin Contents From 30 Batches Produced in China.

In our present study, the standard chemicals of triacetin were purified by reverse-phase and normal-phase semi-preparative high-performance liquid chromatography (HPLC), and 1H NMR and 13C NMR were employed to determine the purity and structure of triacetin. Moreover, a simple and rapid HPLC-photodiode array (PDA) method was developed to determine the contents of triacetin in 30 batches from different suppliers. The chromatographic separation was performed on a Phenomenex Gemini-NX C18 column (250×4.6mm, 5μm) using a gradient elution system of water and acetonitrile (contained 0.1% of formic acid) solution with a flow rate of 1.0mL/min at 30°C at 210nm. Sample preparation method is rapid and energy efficient, and the obtained sample have a good purity. Validation shows good specificity, linearity (R2=0.9995), precision, stability, repeatability (% RSD<2.80) and the average recovery (99.72%) of triacetin. The content of triacetin in most samples is concentrated in 94-97%. This developed approach is simple, rapid, accurate and can be used to quickly determine the purity and the content of triacetin in plasticizers and filter plugs.

LC-MS/MS Estimation of the Anti-Cancer Agent Tandutinib Levels in Human Liver Microsomes: Metabolic Stability Evaluation Assay.

PurposeTandutinib (MLN518 or CT 53518) (TND) is a novel, oral, small-molecule inhibitor of type III receptor tyrosine kinases utilized for the treatment of acute myeloid leukemia (AML).Materials and MethodsIn silico prediction of hepatic drug metabolism for TND was determined using the StarDrop® WhichP450™ module to confirm its metabolic liability. Second, an efficient and accurate LC-MS/MS method was established for TND quantification to evaluate metabolic stability. TND and entrectinib (ENC) (internal standard; IS) were resolved using an isocratic elution system with a reversed stationary phase (C8 column).ResultsThe established LC-MS/MS method exhibited linearity (5–500 ng/mL) with r2 ≥0.9999 in the human liver microsomes matrix. The method sensitivity was indicated by the limit of quantification (3.8 ng/mL), and reproducibility was revealed by inter- and intraday precision and accuracy (below 10.5%). TND metabolic stability estimation was calculated using intrinsic clearance (22.03 µL/min/mg) and in vitro half-life (29.0 min) values.ConclusionTND exhibited a moderate extraction ratio indicative of good bioavailability. According to the literature, the approach developed in the present study is the first established LC-MS/MS method for assessing TND metabolic stability.

Extraction, isolation and characterization of Bauhinia variegata flower

Background: Bauhinia variegata Linn. is a native plant of Asia and China. B. variegata is found in tropical regions of the world. It belongs to family Leguminosae. It is used for diarrhea, hemorrhoids, constipation, piles, edema, leprosy, wounds, tumors, etc. Objective: The objective of the present study was to perform extraction of B. variegata flower and isolation of active constituents from the extract. Materials and Methods: The ethanolic extraction of B. variegata flower was performed using the Soxhlet apparatus. The isolation of active constituents from the extract was performed using chromatographic techniques. In column chromatographic studies, n-hexane- [dichloromethane (DCM)] (2:8) was used as an eluting system and further purified through thin layer chromatography (TLC). Compound A and B were isolated through chromatographic techniques, then the molecular formula and characterization of these compounds were carried out with mass and infrared (IR) spectral analysis. Results and Discussion: The percentage yield of B. variegata ethanolic extract (BVE) was found to be 20.8% w/w. The different fractions were F1 having 12.5 grams with n-hexane, F2 (17.1 grams) with CH2Cl2, F3 (21.2 grams) with EtOAc, and F4 (13.4 grams) with EtOH. Compound A and B were isolated from the solvent fractions of n-hexane-DCM (2:8) and EtOAc-DCM (1:9), respectively. The compound A was characterized as 3-hydroxy-6-methoxy-2-phenyl-4H-chromen-4-one. The compound B was characterized as 3-hydroxy-6-methyl-2-phenyl-4H-chromen-4-one. Conclusion: Thus, B. variegata flowers possess active components that need to identify their biological activities.

Qualitative analysis of cetomacrogol creams by thin-layer chromatography–flame ionization detection (TLC–FID)

Buffered cetomacrogol cream has been described as the cause of iatrogenic allergic contact dermatitis, while patch testing with all ingredients was, in most cases, unable to identify the sensitizing culprit. Several hypotheses had been put forward, among which the formation of a new allergen by interaction of some of the ingredients, so-called ‘compound allergy’. In order to investigate this hypothesis, a method for the qualitative analysis of cetomacrogol creams, using thin-layer chromatography with flame ionization detection (TLC–FID), is presented. All cetomacrogol cream components, i.e., a preservative and excipients were completely separated. A two-step elution system was used to separate the analytes on the Chromarods: in the first step we separated and focussed the paraffins and cetostearyl alcohol with the use of hexane-methanol–methyl tert-butyl ether (100:3:6, v/v). After drying, the same rod was then redeveloped using methanol to resolve sorbic acid from cetomacrogol 1000, whereupon detection of the cream components could be performed by direct flame ionization detection on the Chromarods. The developed method was then applied for the analysis of commercial non-buffered and buffered cetomacrogol cream samples. No newly formed allergen could be detected, thus excluding ‘compound allergy’. This method proved to be simple, cheap, and fast, enabling the separation of the auxiliary substances present in cetomacrogol cream.

IDENTIFIKASI DAN STUDI POLA FRAGMENTASI JAMU TERKONFIRMASI FENILBUTAZON MENGGUNAKAN LIQUID CHROMATOGRAPHY MASS SPECTROSCOPY (LCMS)

The Jamu is one type of traditional herbal medicine that still popular among the community until now. In twenty years often found the jamu containing synthetic drugs. Drug chemical which is commonly added to this traditional herbal medicine include the class of drugs of NAISD’s namely phenylbutazone. This study was conducted to determine the possibility of presence of phenylbutazone in the Jamu. The beginning of this study by initial screening using Thin Layer Chromatography (TLC) method. The results showed that 4 of the 6 samples of the positive contain phenylbutazone based TLC method. These 4 samples were then analyzed using the Liquid Chromatography Mass Spectroscopy (LCMS) method. The stationary phase is Octadecyl Silica (C18) and the Mobile Phase are ultrapure water motion: acetonitrile (90 : 10% v/v) with isocratic elution system and positive ion mode. Based on LCMS chromatogram data, the retention time (Rf) of phenylbutazone is 1.89 minutes. The retention time of samples same with the retention time of phenylbutazone. The next approach, based on the fragmentation pattern of Mass Spectroscopy. Based on mass spectrum data, there are 2 formed fragmentation paths. The first pattern is the fragmentation process of phenylbutazone (309 m / z), forming 2-butylmalonaldehyde (129 m / z) and 1,2-diphenylhydrazine (184 m / z). The second fragmentation pattern was obtained by the Indol 3-H pathway, producing 3-butylidene-3H-indol-2-olate fragments (188 m / z) and 3H-indole-2,3-diide (114 m / z). Based on the analysis using the LCMS method, four samples containing phenylbutazone, Samples A, C, D and E. The four samples should not be allowed to be traded and consumed for violating the Regulation of the Minister of Health of the Republic of Indonesia No 006 of 2012chapter 33 and chapter 37 on the industry and traditional medicine business that traditional medicine is prohibited to contain chemicals from the isolation or synthetic drug efficacious.

Simultaneous Determination of 34 Amino Acids in Tumor Tissues from Colorectal Cancer Patients Based on the Targeted UHPLC-MS/MS Method.

A targeted ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was established and validated for the simultaneous determination of 34 amino acids in tissue samples from colorectal cancer (CRC) patients. The chromatographic separation was achieved on an Agilent ZORBAX SB-C18 column (3.0 × 150 mm, 5 μm) with a binary gradient elution system (A, 0.02% heptafluorobutyric acid and 0.2% formic acid in water, v/v; B, methanol). The run time was 10 min. The multiple reaction monitoring mode was chosen with an electrospray ionization source operating in the positive ionization mode for data acquisition. The linear correlation coefficients were >0.99 for all the analytes in their corresponding calibration ranges. The sample was pretreated based on tissue homogenate and protein precipitation with a 100 mg aliquot sample. The average recovery and matrix effect for 34 amino acids and 3 internal standards were 39.00%∼146.95% and 49.45%∼173.63%, respectively. The intra- and interday accuracy for all the analytes ranged from −13.52% to 14.21% (RSD ≤8.57%) and from −14.52% to 12.59% (RSD ≤10.31%), respectively. Deviations of stability under different conditions were within ±15% for all the analytes. This method was applied to simultaneous quantification of 34 amino acids in tissue samples from 94 CRC patients.