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Advances in Non-Targeted/Targeted Methods for Quality Assurance of Food and Botanical Dietary Supplements. 食品和植物性膳食补充剂质量保证的非靶向/靶向方法研究进展
IF 1.7 Pub Date : 2026-01-10 DOI: 10.1093/jaoacint/qsag004
Mengliang Zhang, Jianghao Sun
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引用次数: 0
Rapidly and simultaneously determining the total polysaccharides content, total flavonoids content and antioxidant activity in Polygonatum cyrtonema Hua by NIR spectroscopy and chemometrics. 采用近红外光谱和化学计量学方法,快速测定了黄精中总多糖含量、总黄酮含量和抗氧化活性。
IF 1.7 Pub Date : 2026-01-10 DOI: 10.1093/jaoacint/qsag003
Xuanxuan Fan, Xian Hu, Weiting Wang, Naifu Chen, Naidong Chen, Jingwen Hao, Le Yan, Jun Xiao, Yu Zhong, Manman Zhang, Zhengjun Xie

Background: Polygonatum cyrtonema Hua (PCH) is widely used in traditional Chinese medicine. The components and activity are important for simultaneously quality evaluation in PCH.

Objective: This study aimed to establish an accurate, rapid and comprehensive quality evaluation method by near-infrared spectroscopy (NIR) for forecasting the total polysaccharides content (TPsC), total flavonoids content (TFC), and antioxidant activity (AA) in PCH.

Methods: PCH samples were used to establish model of TPsC, TFC, and AA in PCH by NIR combined with partial least squares regression (PLS). To enhance the accuracy of the models and remove non-essential variables, we used multiple spectral preprocessing methods and multiple chemometrics to analyze the processed full spectrum, such as competitive adaptive reweighted sampling-partial least squares regression (CARS-PLS), moving window-partial least squares regression (MW-PLS), and interval random frog-partial least squares regression (iRF-PLS). The remaining samples were used to complete external validation.

Results: Satisfactory prediction results of PLS models combined with chemometrics were obtained. The optimal chemometric of TPsC, TFC, and AA selected 157, 105, and 134 variables, respectively. For the TPsC, TFC, and AA models, which incorporated optimal spectral preprocessing and chemometric, the root mean square error of calibration (RMSEC, %) values were 0.743, 0.069, and 0.136, while the R2 values were above 0.95. The root mean square error of prediction (RMSEP, %) for these models were 1.00, 0.074, and 0.153, while R2 values were above 0.90. The remaining 20 samples were used to complete external validation, which confirmed the preeminent comparability and ability of the proposed method.

Conclusion: NIR combined with chemometrics provide an effective, fast, and nondestructive approach for evaluating quality via multiple indicators of PCH and provides a meaningful reference for evaluation of herbal medicine quality.

Highlights: NIR combined with chemometrics offers an effective, quick, nondestructive way to evaluate herbal medicine quality with multiple indicators.

背景:黄精(Polygonatum cytonema Hua, PCH)是一种应用广泛的中药。PCH的组分和活度是同时评价PCH质量的重要指标。目的:建立一种准确、快速、全面的近红外光谱(NIR)质量评价方法,预测PCH中总多糖(TPsC)、总黄酮(TFC)含量和抗氧化活性(AA)。方法:采用近红外结合偏最小二乘回归(PLS)建立PCH中TPsC、TFC和AA的模型。为了提高模型的准确性和去除非必要变量,我们采用了竞争自适应重加权抽样-偏最小二乘回归(CARS-PLS)、移动窗口-偏最小二乘回归(MW-PLS)和区间随机青蛙-偏最小二乘回归(iRF-PLS)等多种光谱预处理方法和多种化学计量学对处理后的全光谱进行分析。剩余样品用于完成外部验证。结果:PLS模型结合化学计量学获得了满意的预测结果。TPsC、TFC和AA的最佳化学计量学分别选择了157、105和134个变量。采用最佳光谱预处理和化学计量学的TPsC、TFC和AA模型的校正均方根误差(RMSEC, %)分别为0.743、0.069和0.136,R2均在0.95以上。这些模型的预测均方根误差(RMSEP, %)分别为1.00、0.074和0.153,R2值均在0.90以上。剩余的20个样本完成外部验证,证实了所提方法具有优异的可比性和能力。结论:近红外联合化学计量学为中药多指标质量评价提供了一种有效、快速、无损的方法,为中药质量评价提供了有意义的参考。重点:近红外光谱与化学计量学相结合,提供了一种有效、快速、无损的多指标草药质量评价方法。
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引用次数: 0
Preparation of functionalized immunomagnetic beads and their application in the detection of AFB1 in Corn, Oat, and Peanut. 功能化免疫磁珠的制备及其在玉米、燕麦和花生中AFB1检测中的应用
IF 1.7 Pub Date : 2026-01-09 DOI: 10.1093/jaoacint/qsag002
Huihui Hao, Mengke Wu, Xiaolei Zhao, Jianjun Li, Jinxing He

Background: Aflatoxin B1 (AFB1) is widespread in various kinds of food and poses a serious threat to health when it enters the human body via the food chain.

Objective: This study aimed to establish a rapid and sensitive method for the detection of AFB1 and then combine it with HPLC for the quantitative analysis of AFB1 content.

Method: In this work, hydrothermal and sol-gel methods were employed to prepare aminoated magnetic nanoparticles featuring strong magnetism and excellent dispersion. Magnetic nanoparticles bound with antibodies can specifically capture AFB1 and then be combined with HPLC for the quantitative analysis of AFB1 content. Moreover, the key factors influencing the extraction efficiency, such as buffer type, adsorption time, elution time and volume, were optimized.

Results: The samples were spiked with AFB1 at low, medium, and high concentration levels of 5.0, 10.0, and 20.0 μg/kg, respectively. The established method exhibits good linearity within the range of 0.1-100 µg/kg, and the detection limit is as low as 0.09 µg/kg (S/N = 3). The recoveries in real samples ranged from 79.33% to 111.51%, with all relative standard deviations (RSDs) being less than 11.16% (n = 3 for each level).

Conclusions: In conclusion, a rapid and efficient analytical method for detecting AFB1 in corn, peanut and oat based on functionalized immunomagnetic beads (IMBs) coupled with HPLC-FLD have been successfully developed .The method exhibited excellent sensitivity, featuring a low method detection limit of 0.09 μg/kg, high accuracy, and remarkable precision. In comparison to traditional sample preparation techniques, this IMBs-based approach presents substantial advantages in terms of simplicity, rapidity (completing sample processing within approximately 30 minutes), and cost-effectiveness.

Highlights: The IMBs possess excellent magnetic separation capabilities, which effectively streamline the pre-processing steps.

背景:黄曲霉毒素B1 (AFB1)广泛存在于各种食品中,通过食物链进入人体后对人体健康构成严重威胁。目的:建立一种快速、灵敏的AFB1检测方法,并结合高效液相色谱法对AFB1含量进行定量分析。方法:采用水热法和溶胶-凝胶法制备了磁性强、分散性好的氨基化磁性纳米颗粒。与抗体结合的磁性纳米颗粒可以特异性捕获AFB1,然后结合HPLC进行AFB1含量的定量分析。并对影响萃取效率的缓冲液类型、吸附时间、洗脱时间和体积等关键因素进行了优化。结果:AFB1在低、中、高浓度分别为5.0、10.0、20.0 μg/kg。建立的方法在0.1 ~ 100µg/kg范围内线性良好,检出限低至0.09µg/kg (S/N = 3)。实际样品的加样回收率为79.33% ~ 111.51%,各水平的相对标准偏差(rsd)均小于11.16% (n = 3)。结论:成功建立了一种基于功能化免疫磁珠(IMBs) - HPLC-FLD的快速、高效检测玉米、花生和燕麦中AFB1的方法。该方法灵敏度高,方法检出限为0.09 μg/kg,准确度高,精密度显著。与传统的样品制备技术相比,这种基于imbs的方法在简单、快速(大约30分钟内完成样品处理)和成本效益方面具有实质性优势。亮点:imb具有出色的磁分离能力,有效地简化了预处理步骤。
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引用次数: 0
"Optimization of Extraction Buffer Conditions for Improved Heparin Affinity-Based Quantification of Bovine Lactoferrin in Various Dairy Products". 基于肝素亲和度的奶牛乳铁蛋白定量提取缓冲条件优化
IF 1.7 Pub Date : 2026-01-08 DOI: 10.1093/jaoacint/qsag001
Tomoaki Fukushima, Keisuke Yamamoto, Yukino Sato, Tatsuya Nojima, Mirei Odaka, Norichika Nishida, Hiroshi Ochi

Background: Bovine lactoferrin (bLF) has been supplemented to a variety of foods, particularly dairy products, to promote human health. Reliable analytical methods are required for accurate quantification of bLF in foods. Isolation of bLF from foods using a heparin column and quantification via HPLC have been reported (heparin method); however, its applicability remains limited.

Objective: To develop a broadly applicable method for determining bLF in dairy products by optimizing the extraction buffer conditions used in the heparin method.

Method: bLF was spiked into different dairy products and extracted using buffers with or without additives, such as urea, sodium chloride, EDTA, and Tween 20. These additives reduced interactions between bLF and dairy components and improved the interaction of bLF to the heparin column, with the buffer pH adjusted to 6.0. The extracted bLF was isolated using a heparin column and quantified via C4 reversed-phase HPLC using an external standard calibration curve.

Results: The optimized extraction buffer improved bLF recovery rates across all tested dairy product matrices.

Conclusions: An improved extraction buffer was developed that enhanced the applicability of bLF quantification using the heparin method. This optimized extraction buffer enabled reliable recovery of bLF from diverse and complex dairy product matrices.

Highlight: The optimized extraction buffer significantly improves the recovery and quantification of bLF in various dairy products, providing a versatile solution for accurate bLF analysis.

背景:牛乳铁蛋白(bLF)已被添加到各种食品中,特别是乳制品中,以促进人类健康。需要可靠的分析方法来准确定量食品中的溴代酚。利用肝素色谱柱分离食品中的bLF并通过高效液相色谱(HPLC)进行定量的报道(肝素法);然而,它的适用性仍然有限。目的:通过优化肝素法提取缓冲液条件,建立一种广泛适用的乳制品中bLF的测定方法。方法:将bLF添加到不同的乳制品中,用添加或不添加尿素、氯化钠、EDTA和Tween 20等添加剂的缓冲液提取。这些添加剂降低了bLF与乳制品组分的相互作用,提高了bLF与肝素柱的相互作用,缓冲液pH调整为6.0。提取的bLF采用肝素柱分离,C4反相高效液相色谱法采用外标曲线定量。结果:优化后的萃取缓冲液提高了所有测试乳制品基质的bLF回收率。结论:开发了一种改进的提取缓冲液,提高了肝素法定量bLF的适用性。这种优化的提取缓冲液能够从各种复杂的乳制品基质中可靠地回收bLF。重点:优化后的萃取缓冲液显著提高了各种乳制品中bLF的回收率和定量,为bLF的准确分析提供了一个通用的解决方案。
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引用次数: 0
Efficient Catalysis Based on ZnFe2O4/BSN-rGO Nanoparticles for Chlorogenic Acid Detection. 基于ZnFe2O4/BSN-rGO纳米颗粒的绿原酸检测高效催化
IF 1.7 Pub Date : 2025-12-31 DOI: 10.1093/jaoacint/qsaf111
Hua Chen, Hehua Zhang, Xin Gu, Hongzhi Pan

Background: Chlorogenic acid (CGA), a plant-derived phenolic compound, possesses antioxidant, anti-inflammatory, and antimicrobial activities. Accurate quantification is essential due to its therapeutic value and potential toxicity at high doses.

Objective: To develop a rapid, sensitive electrochemical method for CGA detection using spinel-based catalysts with improved conductivity.

Methods: An electrochemical sensing platform was constructed based on a nanocomposite of ZnFe2O4 and boron, sulfur and nitrogen co-doped reduced graphene oxide (BSN-rGO). This hybrid material effectively addresses the intrinsic poor conductivity of spinel oxides by leveraging the synergistic effect between the catalytic activity of ZnFe2O4 and the superior charge transport capability of BSN-rGO. The resulting sensor enabled rapid and quantitative analysis of CGA within two minutes.

Results: The sensor showed a linear detection range of 1 × 10-4 -1 × 10-1 M and a low detection limit of 1 × 10-6 M (S/N = 3). Adding standard recovery reached 97.48% to 100.75%, with RSDs of 1.02% to 2.95%, indicating excellent accuracy and reproducibility in complex matrices.

Conclusion: The proposed method enables rapid and reliable CGA detection in natural products, offering potential for pharmaceutical and food analysis.

Highlights: BSN-rGO integration enhances spinel conductivity and boosts electrochemical sensing performance.

背景:绿原酸(CGA)是一种植物源性酚类化合物,具有抗氧化、抗炎和抗菌活性。由于其治疗价值和高剂量时的潜在毒性,准确的定量是必不可少的。目的:建立一种快速、灵敏、电导率提高的尖晶石基催化剂检测CGA的电化学方法。方法:采用纳米复合材料ZnFe2O4与硼、硫、氮共掺杂的还原性氧化石墨烯(BSN-rGO)构建电化学传感平台。该杂化材料利用ZnFe2O4的催化活性和BSN-rGO优异的电荷输运能力之间的协同效应,有效地解决了尖晶石氧化物固有的导电性差的问题。由此产生的传感器可以在两分钟内快速定量分析CGA。结果:该传感器线性检测范围为1 × 10-4 -1 × 10-1 M,低检出限为1 × 10-6 M (S/N = 3)。加标回收率为97.48% ~ 100.75%,rsd为1.02% ~ 2.95%,在复杂基质中具有良好的准确度和重现性。结论:该方法能够快速、可靠地检测天然产物中的CGA,为制药和食品分析提供了潜在的应用前景。亮点:BSN-rGO集成增强尖晶石导电性,提高电化学传感性能。
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引用次数: 0
Research on the Preparation and Application of Lyoprotectants for Hanseniaspora uvarum BF-345 Derived from Indigenous Grape Juice. 本土葡萄汁源发酵菌BF-345冻干保护剂的制备及应用研究。
IF 1.7 Pub Date : 2025-12-23 DOI: 10.1093/jaoacint/qsaf110
Qiao Song, Jiawei Yang, Tao Peng, Hongmei Tong, Xiaolun Zhou, Jing Wang

Background: The extensive use of imported commercial Saccharomyces cerevisiae active dry powder preparations has led to a serious homogenization of wine products in China.

Objective: In recent years, there has been growing interest in developing and applying indigenous non-Saccharomyces yeasts to enhance the diversity and distinctive character of wines.

Methods: In this study, a promising indigenous strain, Hanseniaspora uvarum BF-345, previously isolated from local grape juice, was investigated due to its desirable fermentation properties, robust stress tolerance, and high β-glucosidase activity. Response surface methodology (RSM) was employed to optimize the composition of a cryoprotectant formulation for the vacuum freeze-drying of BF-345.

Results: The optimal protectant formulation was determined as follows: 5.63% skim milk powder, 12.89% maltitol, and 7.02% sucrose. Under these conditions, the viability of the freeze-dried cells reached 85.2%, representing a 27.3% increase compared to the pre-optimized formulation.

Conclusion: The prepared active dry yeast complied with the Chinese national standard GB/T 20886.2-2021, and maintained a viability above 70% after 6 months of sealed storage at 4 °C.

Highlights: In micro-vinification trials with Cabernet Sauvignon, the fermentation system inoculated with the freeze-dried powder exhibited higher yeast cell counts and more favorable fermentation kinetics than that inoculated with a liquid seed culture.

背景:进口商业酿酒酵母活性干粉制剂的广泛使用导致中国葡萄酒产品同质化严重。目的:近年来,人们对开发和应用本土非酵母菌的兴趣日益浓厚,以增强葡萄酒的多样性和独特性。方法:在这项研究中,研究了一种有前途的本土菌株,汉斯iaspora uvarum BF-345,该菌株先前从当地葡萄汁中分离出来,由于其良好的发酵特性,强大的耐受性和高β-葡萄糖苷酶活性。采用响应面法(RSM)对BF-345真空冷冻干燥用冷冻保护剂配方进行优化。结果:确定最佳保护剂配方为:5.63%脱脂奶粉、12.89%麦芽糖醇、7.02%蔗糖。在此条件下,冻干细胞的存活率达到85.2%,比优化前的配方提高了27.3%。结论:制备的活性干酵母符合中国国家标准GB/T 20886.2-2021,在4℃密封保存6个月后,活性维持在70%以上。在赤霞珠的微发酵试验中,用冻干粉接种的发酵系统比用液体种子培养接种的发酵系统表现出更高的酵母细胞计数和更有利的发酵动力学。
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引用次数: 0
Correction to: Development of Psilocybe Mushroom Species Reference Material-Cultivation Parameters and Chemical Profiles. 修正:裸盖菇品种参考材料的开发-培养参数和化学特征。
IF 1.7 Pub Date : 2025-12-19 DOI: 10.1093/jaoacint/qsaf104
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引用次数: 0
Evaluating physicochemical and antioxidant properties of farm-fresh and branded honey from bangladesh using UHPLC-DAD and multivariate analysis. 利用UHPLC-DAD和多变量分析评价孟加拉国农场新鲜蜂蜜和品牌蜂蜜的理化和抗氧化性能。
IF 1.7 Pub Date : 2025-12-18 DOI: 10.1093/jaoacint/qsaf109
Jahangir Alam, Manobendra Nath Mohanta, Tamanna Tasnim, Md Azmain Faique, Saidul Islam, Md Athar Ishtiyaq, Mahmudul Bhuiyan, Md Yeasin Prodhan, Md Najim Uddin, Khalid Ali Khan, Md Atikul Islam

Background: The market demand of honey in Bangladesh is raising day by day, but there are concerns about the quality of both farm-fresh and branded honey which are available in the market.

Objectives: This study evaluates the physicochemical and antioxidant properties of farm-fresh and branded honey from the Bangladeshi market.

Methods: The moisture content (MC) and total soluble solid (°Bx) were determined by refractometer; the electrical conductivity (EC) determined by digital conductivity meter; total phenol (TP), total flavonoid (TF) and HMF were measured by Folin-Ciocalteu, aluminum chloride and ultra high performance liquid chromatography-diode array detector (UHPLC-DAD) respectively; and antioxidant capacity was assessed by 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging method.

Results: The results revealed that, farm-fresh honey samples exhibited greater content of moisture (>20%), (329.63 ± 1.63 to 986.05 ± 4.81 mg GAE/kg), total flavonoid (95.09 ± 4.99 to 454.73 ± 2.45 mg QE/kg) and antioxidant capacity (IC50 value of DPPH, 47.91 ± 0.19 to 73.66 ± 0.30 mg/mL) compared to branded honey samples (p < 0.05). The HMF amount was varied from 0.82 ± 0.00 to 241.24 ± 2.41 (mg/kg), with forty-two percent (42%) of branded honey samples exceeding both the Bangladesh Standards and Testing Institution (BSTI) and Codex Alimentarius standard limit (80 mg/kg), whereas all farm-fresh honey samples were within the acceptable limit. In multivariate analysis, the principal component (PC1 & PC2) analysis explains 81.30% of the total variance.

Conclusions: Based on compliance with BSTI and Codex HMF limit, farm fresh honey samples meet the acceptable HMF threshold, whereas 42% of branded honey exceeded the standard limit.

Highlights: Analyzed 45 honey samples (farm-fresh, national, international) for physicochemical parameters; Farm-fresh honey exhibited higher moisture, phenolics, flavonoids and antioxidant activity; 42% of branded honeys exceeded HMF standard limits (BSTI and Codex); all farm-fresh samples were within permissible limit.

背景:孟加拉国市场对蜂蜜的需求日益增加,但市场上可用的农场新鲜蜂蜜和品牌蜂蜜的质量都令人担忧。目的:本研究评价了孟加拉国市场上农场新鲜蜂蜜和品牌蜂蜜的物理化学和抗氧化性能。方法:用折光仪测定水分(MC)和总可溶性固形物(°Bx);电导率(EC)由数字电导率仪测定;用Folin-Ciocalteu、氯化铝和超高效液相色谱-二极管阵列检测器(UHPLC-DAD)分别测定总酚(TP)、总黄酮(TF)和HMF;采用2,2′-二苯基-1-吡啶酰肼(DPPH)自由基清除法测定抗氧化能力。结果:与品牌蜂蜜相比,农场新鲜蜂蜜的水分含量(bbb20 %),(329.63±1.63 ~ 986.05±4.81 mg GAE/kg),总黄酮含量(95.09±4.99 ~ 454.73±2.45 mg QE/kg)和抗氧化能力(DPPH IC50值,47.91±0.19 ~ 73.66±0.30 mg/mL)均高于品牌蜂蜜(p)。根据对BSTI和食品法典委员会HMF限值的遵守情况,农场新鲜蜂蜜样品符合可接受的HMF阈值,而42%的品牌蜂蜜超过标准限值。亮点:分析了45份蜂蜜样品(农场新鲜、国内、国际)的理化参数;农场新鲜蜂蜜具有较高的水分、酚类物质、类黄酮和抗氧化活性;42%的品牌蜂蜜超过HMF标准限值(BSTI和食品法典委员会);所有农场新鲜样品均在允许范围内。
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引用次数: 0
Development and Comparison of Stability-Indicating HPLC and UPLC Methods for the Determination and Validation of Pantoprazole Sodium Impurities. 稳定性指示高效液相色谱法与超高效液相色谱法测定泮托拉唑钠杂质及验证方法的建立与比较
IF 1.7 Pub Date : 2025-12-12 DOI: 10.1093/jaoacint/qsaf107
Aysen Kurt Cucu, Engin Ozturk

Background: Pantoprazole is a selective proton pump inhibitor used in its sodium form in pharmaceuticals. Determining its impurities is crucial for drug purity and safety.

Objective: This study aims to develop a faster impurity analysis method than the European Pharmacopoeia (EP) method.

Methods: An Agilent Zorbax SB Phenyl (250 × 4.6 mm, 5.0 µm) column was used for HPLC, and a Restek Ultra Biphenyl (100 × 2.1 mm, 3.0 µm) column for UPLC. A 65:35 (v/v) potassium dihydrogen/dipotassium hydrogen phosphate buffer (pH 7.40)-acetonitrile mixture was the mobile phase. Analyses were performed at 290 nm with flow rates of 1.0 mL/min (HPLC) and 0.25 mL/min (UPLC). Injection volumes were 20 µL (HPLC) and 3.5 µL (UPLC).

Results: The analysis times of the impurity determination methods developed are 15 min shorter for the HPLC method and 30 minutes shorter for the UPLC method compared to the EP method. The developed methods were validated according to ICH guidelines, demonstrating high linearity (R2 > 0.99 over the concentration range of 0.03-2.27 µg/mL), accuracy with mean recoveries ranging from 95.0% to 105.0%, and precision with intra- and inter-day RSD values below 2%. Forced degradation studies under acidic, alkaline, oxidative, thermal, and photolytic conditions confirmed that the methods are stability-indicating, with the ability to separate all major degradation products of pantoprazole sodium.

Conclusion: The method was validated according to International Conference on Harmonisation (ICH) guidelines, and forced degradation studies were also performed. This study presents optimized HPLC and UPLC methods that allow faster impurity analysis and detection of an additional non-compendial impurity, Pantoprazole-N-oxide.

Highlights: As a result of shorter analysis times, a significant cost reduction has also been proven with numerical data. In addition, a performance-cost comparison of two different analytical technologies was made.

背景:泮托拉唑是一种选择性质子泵抑制剂,在药物中以钠形式使用。确定其杂质对药物纯度和安全性至关重要。目的:建立一种比欧洲药典(EP)方法更快的杂质分析方法。方法:高效液相色谱柱为Agilent Zorbax SB Phenyl (250 × 4.6 mm, 5.0µm),高效液相色谱柱为Restek Ultra Biphenyl (100 × 2.1 mm, 3.0µm)。流动相为65:35 (v/v)二氢钾/磷酸氢二钾缓冲液(pH 7.40)-乙腈混合物。在290 nm下,流速为1.0 mL/min (HPLC)和0.25 mL/min (UPLC)。进样量分别为20µL (HPLC)和3.5µL (UPLC)。结果:HPLC法和UPLC法的分析时间分别比EP法缩短了15 min和30 min。该方法在0.03 ~ 2.27 μ g/mL范围内具有良好的线性关系(R2为0.99),平均加样回收率为95.0% ~ 105.0%,日内和日间RSD值低于2%。在酸性、碱性、氧化性、热和光解条件下的强制降解研究证实,该方法具有稳定性,能够分离泮托拉唑钠的所有主要降解产物。结论:该方法根据国际统一会议(ICH)指南进行了验证,并进行了强制降解研究。本研究提出了优化的HPLC和UPLC方法,可以更快地进行杂质分析和检测另一种非药典杂质Pantoprazole-N-oxide。亮点:由于分析时间缩短,数值数据也证明了显著的成本降低。此外,还对两种不同分析技术的性能成本进行了比较。
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引用次数: 0
Single-Chain Variable Fragment Based Dot Blot, Single and Multiple Assays for Rapid SARS-CoV-2 Diagnostics. 基于单链可变片段点印迹、单次和多次快速诊断SARS-CoV-2的试验
IF 1.7 Pub Date : 2025-12-06 DOI: 10.1093/jaoacint/qsaf105
Dilek Çam Derin, Enes Gültekin, Irmak İçen Taşkın, Muhammed Dündar, Barış Otlu

Background: Antigenic detection is reliably utilized in rapid diagnostic tests and provides a significant time advantage during pandemics and epidemics. Therefore, the rapid detection of viral infections is of great importance and will remain crucial in the future. The SARS-CoV-2 outbreak, which resulted in severe losses, is the most recent example of this necessity. Among rapid diagnostic tests, lateral flow assays (LFAs) are the most practical and do not require specialized equipment, typically being developed using antibody pairs.

Objective: This study aimed to recombinantly produce a single-chain variable fragment (scFv) specific to the SARS-CoV-2 spike receptor-binding domain (sRBD) and to employ it in the development of lateral flow assays (LFAs) utilizing both antibody and aptamer pairs and an aptamer cocktail.

Methods: Gold nanoparticles were employed as labeling agents, while both the scFv and full length forms of CR3022, along with aptamers specific to the S and N proteins, were utilized in a sandwich assay format.

Results: scFv was produced at a higher concentration and biologically active. It demonstrated effective viral detection in single LFA, Dot Blot Assay (DBA), and multiplex LFA. While single LFA successfully detected only the synthetic target, DBA and multiplex LFA selectively identified the virus in nasopharyngeal and oropharyngeal swab samples.

Conclusion: Findings highlight the differences and effectiveness of using scFv in combination with other capture agents and different assay principles for the development of cost-effective and rapid diagnostic tests.

Highlights: scFvs exhibit variable binding in sandwich assays depending on the combinations employed. When used in combination with an aptamer cocktail, scFvs demonstrate enhanced target binding, which is shown for the first time in this study. The use of multiple testing strategies enables a more effective viral diagnosis.

背景:抗原检测可靠地用于快速诊断测试,并在大流行和流行病期间提供了显著的时间优势。因此,快速检测病毒感染是非常重要的,在未来仍将是至关重要的。造成严重损失的SARS-CoV-2疫情是这种必要性的最新例子。在快速诊断测试中,横向流动测定法(LFAs)是最实用的,不需要专门的设备,通常使用抗体对开发。目的:本研究旨在重组产生SARS-CoV-2刺突受体结合域(sRBD)特异性单链可变片段(scFv),并将其用于利用抗体和适配体对以及适配体鸡尾酒开发侧流分析(LFAs)。方法:使用金纳米颗粒作为标记剂,同时使用scFv和全长形式的CR3022,以及特定于S和N蛋白的适配体,在三明治检测格式中使用。结果:scFv的产率较高,具有较高的生物活性。它在单LFA, Dot Blot Assay (DBA)和多重LFA中显示出有效的病毒检测。单个LFA仅能成功检测合成靶点,而DBA和多重LFA能选择性地鉴定鼻咽和口咽拭子样本中的病毒。结论:研究结果突出了scFv与其他捕获剂和不同检测原则联合使用的差异和有效性,以开发具有成本效益的快速诊断测试。重点:scFvs在三明治试验中表现出不同的结合,这取决于所采用的组合。当与适体混合物联合使用时,scFvs表现出增强的靶标结合,这在本研究中是首次得到证实。使用多种检测策略可以更有效地进行病毒诊断。
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Journal of AOAC International
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