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Development of a green and rapid ethanol-based HPLC assay for aspirin tablets and feasibility evaluation of domestically produced bioethanol in Thailand as a sustainable mobile phase

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GREEN PROCESSING AND SYNTHESIS 2025年第1期14卷

作者： thanayutsiri, thana Mantadilok, Siwa Sapsin, Jiradet Tungwattanaviboon, Thada Wongwatanasin, Jetsada Opanasopit, Praneet Ngawhirunpat, Tanasait Rojanarata, Theerasak Silpakorn Univ Fac Pharm Sanam Chandra Palace Campus Nakhon Pathom 73000 Thailand Silpakorn Univ Fac Pharm Pharmaceut Dev Green Innovat Grp PDGIG Nakhon Pathom 73000 Thailand Thai Ethanol Mfg Assoc Bangkok 10400 Thailand

Despite the growing trend of using ethanol as a greener alternative to hazardous solvents like methanol and acetonitrile in high-performance liquid chromatography (HPLC) analysis, no ethanol-based assay has been developed for aspirin tablets, nor has the potential of domestically produced bioethanol as a mobile phase been explored - until now. This study introduces a novel ethanol-based HPLC assay for aspirin and assesses the feasibility of using locally produced bioethanol. The optimized method, featuring a 40% (v/v) ethanol-water mobile phase adjusted to pH 3.6 with glacial acetic acid, with a 15-cm C18 column at 40 degrees C and UV detection at 237 nm, successfully separated aspirin and its degradation product, salicylic acid, in 5 min. It demonstrated excellent linearity (0.1-0.6 mgmL(-1), r(2) = 0.9997), sensitivity (LOD = 0.01 mgmL(-1), LOQ = 0.03 mgmL(-1)), accuracy, and precision, with no interference from excipients. The method's performance on commercial aspirin tablets aligned with official pharmacopoeial methods. In a case study using fuel-grade bioethanol (>99.5% purity) derived from sugarcane molasses or cassava from local manufacturers in Thailand, chromatographic and analytical results matched those obtained with imported ethanol, validating its effectiveness. This innovative approach not only reduces costs and dependence on imported solvents but also supports local bio-circular-green economies and promotes greener, more sustainable analytical practices.

关键词： ethanol HPLC aspirin domestic sustainability

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A novel, green and affordable semi-quantitative limit test for2-aminobutanol impurity in ethambutol hydrochloride bulk and tablets

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PHARMACIA 2024年 71卷 1-7页

作者： thanayutsiri, thana Patrojanasophon, Prasopchai Opanasopit, Praneet Ngawhirunpat, Tanasait Rojanarata, Theerasak Silpakorn Univ Fac Pharm Dept Pharmacol & Toxicol Sanam Chandra Palace Campus Nakhon Pathom 73000 Thailand

The pharmacopoeial limit test for the impurity 2-aminobutanol (AB) in ethambutol hydrochloride (ETB) relies on florescence measurement using a fluorometer. However, this instrument is often unavailable in many laboratories due to its cost. A novel colorimetric method was therefore developed using genipin, an amine-reactive reagent, to react with AB, producing a blue color. The color intensity was subsequently measured using a more affordable UV-vis spectrophotometer. To overcome matrix effects, the standard addition technique was incorporated. The test effectively detected AB at concentrations as low as 0.125% in ETB, below the compendial limit of 1.0%. The analysis of ETB bulk and tablets showed consistent semi-quantitative results with the USP method. Furthermore, it aligned with green chemistry by eliminating the use of organic solvents and employing safe, naturally derived reagent. In summary, the proposed method provides a reliable, green, and cost-effective alternative for analyzing AB in ETB, ensuring compliance with pharmacopeial standards.

关键词： limit test 2-aminobutanol ethambutol hydrochloride UV-vis spectrophotometry genipin

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Facile, sensitive and reagent-saving smartphone-based digital image colorimetric assay of captopril tablets enabled by long-pathlength RGB acquisition

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PHARMACIA 2023年第4期70卷 1511-1519页

作者： thanayutsiri, thana Charoenying, Thapakorn Patrojanasophon, Prasopchai Pamornpathomkul, Boonnada Opanasopit, Praneet Ngawhirunpat, Tanasait Rojanarata, Theerasak Silpakorn Univ Pharmaceut Dev Green Innovat Grp PDGIG Fac Pharm Amphoe Muang 73000 Nakhon Pathom Thailand Silpakorn Univ Dept Ind Pharm Fac Pharm Amphoe Muang 73000 Nakhon Pathom Thailand

A new assay of captopril (CTP) tablets was developed based on digital image colorimetry using Ellman's reagent. For the first time, a facile technique of increasing the analytical path was applied in this work to enhance the sensitivity. For this purpose, the reaction solutions were photographed using a smartphone while they were contained in two 1-cm pathlength cuvettes which were placed side by side. The Red-Green-Blue (RGB) in term of [B/(R+G+B)] was used to plot a standard curve. Compared to using a single cuvette, double cuvettes resulted in more precise analytical signals and better linearity (r(2) of 0.9992). Additionally, CTP could be analyzed at low concentrations (2.5-25 mu M) with LOD of 0.70 mu M and LOQ of 2.13 mu M, thus lowering the reagent consumption. The assay was proven to be valid, and it was greener, faster, and more affordable than the pharmacopeial chromatographic method, thereby suitable for pharmaceutical quality control.

关键词： smartphone digital image colorimetry long pathlength captopril

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Investigating matrix interference in the pharmacopeial limit test for aluminum in citric acid: a re-examination, for revision of the method

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PHARMACIA 2022年第1期69卷 9-13页

作者： Rojanarata, Theerasak Maithongdee, Kittithat Yuwansri, Nattapong Kaewprasert, Sirada thanayutsiri, thana Phadungcharoen, Noppharat Chinsriwongkul, Akhayachatra Silpakorn Univ Fac Pharm Nakhon Pathom Thailand Newcharoen Pharmaceut LP Pathum Thani Thailand

In many pharmacopoeias, the limit test used for determining the level of aluminum in citric acid labeled for use in the manufacture of dialysate, is based on solvent extraction using 8-hydroxyquinoline and measurement of fluorescence. However, the fluorescence intensity (F.I.) readout from the extract of citric acid samples has been found to be highly dubious, showing low value, and even lower than that of a blank solution. The aim of this work therefore was to examine what effects the matrix has on the test. The comparison of the two standard curves of aluminum solutions in water, against those prepared in citric acid solutions revealed that they differed greatly in terms of slope and y-intercept. In addition, the F.I. values on the plot of the citric acid solution were much lower than that prepared in the water. In another experiment, a decrease in the F.I. of aluminum solution was clearly seen when the co-existing concentration of citric acid was increased. The results inferred that citric acid interfered with the test due to its acidity and metal-chelating capabilities. Based on this evidence, the pharmacopeial limit test for aluminum in citric acid should be revised;otherwise, it could yield results that underestimate aluminum levels and lead to inaccurate conclusions

关键词： aluminum citric acid interference limit test matrix effects

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Rapid and efficient microwave-assisted extraction of Caesalpinia sappan Linn. heartwood and subsequent synthesis of gold nanoparticles

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GREEN PROCESSING AND SYNTHESIS 2023年第1期12卷

作者： thanayutsiri, thana Patrojanasophon, Prasopchai Opanasopit, Praneet Ngawhirunpat, Tanasait Laiwattanapaisal, Wanida Rojanarata, Theerasak Silpakorn Univ Fac Pharm Pharmaceut Dev Green Innovat Grp PDGIG Nakhon Pathom 73000 Thailand Silpakorn Univ Fac Pharm Dept Ind Pharm Pharmaceut Dev Green Innovat Grp PDGIG Nakhon Pathom 73000 Thailand Chulalongkorn Univ Fac Allied Hlth Sci Dept Clin Chem Biosensors & Bioanalyt Technol Cells & Innovat Tes Bangkok 10330 Thailand

Since microwave (MW)-assisted synthesis of gold nanoparticles (AuNPs) using Caesalpinia sappan (CS) extract as both a reducing and stabilizing agent is currently unavailable, a MW-based synthesis protocol was investigated and presented for the first time in this work. In addition, to rapidly prepare the reactant for this purpose, the MW-assisted extraction of CS heartwood was studied. From the optimization experiments, it was found that the extraction using the MW irradiation at 300 W for 3 min produced the extract with high and reproducible brazilin content which could be readily used for the synthesis of AuNPs. Under the optimal synthesis conditions, roughly spherical CS-AuNPs with an average diameter size of 49.6 nm and acceptable 28-day stability were obtained within only 1 min. The resulting CS-AuNPs were capable of selective binding to Fe2+, Fe3+, and Al3+, leading to particle aggregation as well as noticeable change of color and shift of UV-Vis absorption maxima. From these results, CS-AuNPs could be fabricated via this fast, green, and efficient route. Furthermore, their potential application for colorimetric sensing of certain metal ions was preliminarily explored and proposed in this work.

关键词： microwave extraction synthesis gold nanoparticles Caesalpinia sappan Linn

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Rapid synthesis of chitosan-capped gold nanoparticles for analytical application and facile recovery of gold from laboratory waste

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CARBOHYDRATE POLYMERS 2020年 250卷 116983-116983页

作者： thanayutsiri, thana Patrojanasophon, Prasopchai Opanasopit, Praneet Ngawhirunpat, Tanasait Plianwong, Samawadee Rojanarata, Theerasak Silpakorn Univ Fac Pharm Pharmaceut Dev Green Innovat Grp PDGIG Muang 73000 Nakhon Pathom Thailand Burapha Univ Fac Pharmaceut Sci Chon Buri 20131 Thailand

Chitosan-capped gold nanoparticles (CS-AuNPs) were synthesized via a rapid method by exposing 0.75 mL of 10 mM HAuCl4 and 5 mL of 0.1% w/v chitosan solution to microwave irradiation at 125 degrees C for 40 s. The resulting red colloidal solution contained monodispersed, spherical, and stable CS-AuNPs with a mean diameter of 17.8 nm and showed the maximum absorption peak at 520 nm. Owing to the positively charged surface, CS-AuNPs were applied to the assay of the polyanionic compound disodium edetate based on the aggregation induced red-to-blue color change of the particles and the measurement of increased absorbance at 670 nm. The assay was accurate and precise;its results were comparable to those obtained from the titrimetric reference method. Moreover, gold was shown to be easily recovered from CS-AuNPs through alkaline-induced precipitation followed by acid treatment and heating, which provides a facile and efficient means for treatment of laboratory waste.

关键词： Chitosan Gold nanoparticles Microwave Assay Waste treatment

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Improving pharmaceutical analysis by incorporating green chemistry and smartphone technology: The assay for dissolution test of ethambutol tablets as an example

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SUSTAINABLE CHEMISTRY AND PHARMACY 2023年 36卷

作者： thanayutsiri, thana Ruttanakorn, Kanong Laiwattanapaisal, Wanida Fernandez, Raimon Gaspar Rojanarata, Theerasak Silpakorn Univ Fac Pharm Pharmaceut Dev Green Innovat Grp PDGIG Nakhon Pathom 73000 Thailand Silpakorn Univ Fac Pharm Dept Ind Pharm Nakhon Pathom 73000 Thailand Chulalongkorn Univ Fac Allied Hlth Sci Ctr Excellence Biosensors & Bioengn CEBB Dept Clin Chem Bangkok 10330 Thailand Programmer Barcelona 08029 Spain

This work showcases the improvement of the United States Pharmacopeial (USP) assay of ethambutol (ETB) for tablet dissolution test through the incorporation of green chemistry and smartphone technology. In the assay, downscaled extraction was performed in microcentrifuge tubes, utilizing 0.5 mL of chloroform per sample. Instead of spectrophotometric measurements, the extracts were photographed while contained in the closed tubes using a smartphone camera, with white LED serving as the light source. The Red-Green-Blue (RGB) intensities were read out from the image using an RGB Color Detector application. It was found that B/(R + G + B) showed a good linear relationship to the drug concentration over the range of 11.9-118.5 mu g/mL (R2 = 0.9992). The method was accurate (% recovery of 99.98-100.91%), precise (% relative standard deviation of 0.97-1.17%), sensitive (LOD = 3.9 mu g/mL and LOQ = 11.8 mu g/mL), specific and robust. Furthermore, the analysis results obtained were consistent with those obtained through the USP method. The method became greener and safer due to a 20-fold reduction in chloroform consumption and the use of a closed system for signal measurement which prevented the release of solvent vapor into the environment and minimized the analyst's exposure to chemicals. In addition, the method was green with the AGREE score of 0.74, and 9 green shaded sections in the GAPI assessment. Moreover, the data analysis tool in the mobile application developed in this work facilitated rapid and instantaneous calculations of the results. Hence, the assay is feasible for quality control analysis, and helps promote sustainability.

关键词： Assay Green chemistry Smartphone Digital image colorimetry Ethambutol

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Green and facile short path smartphone-based assay for L-proline in dietary supplements using a mild chromogenic reaction combined with simple equipment

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SUSTAINABLE CHEMISTRY AND PHARMACY 2024年 42卷

作者： thanayutsiri, thana Patrojanasophon, Prasopchai Opanasopit, Praneet Ngawhirunpat, Tanasait Chinsriwongkul, Akhayachatra Rojanarata, Theerasak Silpakorn Univ Fac Pharm Dept Pharmacol & Toxicol Sanam Chandra Palace Campus Nakhon Pathom 73000 Thailand Newcharoen Pharmaceut LP Pathum Thani 12120 Thailand

A novel method for determining L-proline content in dietary supplements was developed, relying on a chromogenic reaction between L-proline and isatin. Because of the resulting intense blue color, solutions were transferred to short pathlength cuvettes (0.2 cm) in order to lower the color intensity, eliminating the need for dilution, before being subjected to photography using a smartphone and subsequent Red-Green-Blue (RGB) analysis. The intensity of the red color (R), expressed as log (R-blank/R), was used as an analytical signal, providing good linearity (r(2) = 0.9994) over a concentration range of 0.8 - 8.0 mM. The assay exhibited satisfactory accuracy, precision, and sensitivity (with a limit of detection of 0.24 mM and a limit of quantification of 0.72 mM), without interference from excipients and other 19 essential amino acids. Additionally, the results were consistent with those obtained through the currently used methods, i.e., nonaqueous titration and spectrophotometry using ninhydrin as a chromogenic reagent. Therefore, a green and simple by design analytical method development, marked by the utilization of a mild reaction in a micro-scale (300 mu L), eco-friendlier reagents, reduced solvent usage, and simplified procedures using basic and inexpensive equipment while also minimizing chemical exposure, was effective in overcoming limitations in existing assays for L-proline. Moreover, the use of short pathlength cuvettes for RGB acquisition in smartphone-based digital image colorimetry, introduced for the first time in this work, demonstrates potential for analyzing other substances.

关键词： Short path Smartphone Assay L-proline Dietary supplement Isatin

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