Current Trends and Challenges for Rapid SMART Diagnostics at Point-of-Site Testing for Marine Toxins

In the past twenty years marine biotoxin analysis in routine regulatory monitoring has advanced significantly in Europe (EU) and other regions from the use of the mouse bioassay (MBA) towards the high-end analytical techniques such as high-performance liquid chromatography (HPLC) with tandem mass spectrometry (MS). Previously, acceptance of these advanced methods, in progressing away from the MBA, was hindered by a lack of commercial certified analytical standards for method development and validation. This has now been addressed whereby the availability of a wide range of analytical standards from several companies in the EU, North America and Asia has enhanced the development and validation of methods to the required regulatory standards. However, the cost of the high-end analytical equipment, lengthy procedures and the need for qualified personnel to perform analysis can still be a challenge for routine monitoring laboratories. In developing regions, aquaculture production is increasing and alternative inexpensive Sensitive, Measurable, Accurate and Real-Time (SMART) rapid point-of-site testing (POST) methods suitable for novice end users that can be validated and internationally accepted remain an objective for both regulators and the industry. The range of commercial testing kits on the market for marine toxin analysis remains limited and even more so those meeting the requirements for use in regulatory control. Individual assays include enzyme-linked immunosorbent assays (ELISA) and lateral flow membrane-based immunoassays (LFIA) for EU-regulated toxins, such as okadaic acid (OA) and dinophysistoxins (DTXs), saxitoxin (STX) and its analogues and domoic acid (DA) in the form of three separate tests offering varying costs and benefits for the industry. It can be observed from the literature that not only are developments and improvements ongoing for these assays, but there are also novel assays being developed using upcoming state-of-the-art biosensor technology. This review focuses on both currently available methods and recent advances in innovative methods for marine biotoxin testing and the end-user practicalities that need to be observed. Furthermore, it highlights trends that are influencing assay developments such as multiplexing capabilities and rapid POST, indicating potential detection methods that will shape the future market.

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Method Development and Validation of Degradation Studies of Favipiravir by RP-HPLC

Journal of Pharmaceutical Research International ◽

10.9734/jpri/2021/v33i42a32399 ◽

2021 ◽

pp. 220-228

Author(s):

. Shyamala ◽

Dongamanti Ashok

Keyword(s):

Mobile Phase ◽

Method Development ◽

Hplc Method ◽

Mobile Phase Flow Rate ◽

Rp Hplc ◽

Wide Range ◽

Method Development And Validation ◽

Development And Validation ◽

Analytical Standards ◽

Theoretical Plates

RP HPLC method was developed by this study estimation of favipiravir. This method is developed by Shimadzu LC -2010 HT by using a C18 (250 X 4.6 X mm X 5µ) column in solvents Water(OPA)+ACN (60:40)v/v as mobile phase and the temperature was maintained at 25°C. The mobile phase flow rate of 1ml/min was pumped and sample wavelength was detected at 324 nm by ultraviolet -visible spectrophotometer. The Rt was found 4.453 min. The number of theoretical plates and tailing factor for favipiravir was observed 82651(NLT 2000) and 1.265 (NMT 2). The method was validated for analytical standards such as linearity, accuracy, precision, LOD, LOQ and robustness. LOD and LOQ values were calculated from regression of favipiravir 1.26 and 3.83 µg/ml.The regression equation of validated method for favipiravir is Y=253.5x+1881.In a wide range of 4 to 20 (µg/ml) the linearity was observed. Degradation methods were also performed.

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HPLC METHOD DEVELOPMENT AND VALIDATION FOR AZITHRO-MYCIN IN ORAL SUSPENSION

Journal of Applied Pharmaceutical Sciences and Research ◽

10.31069/japsr.v2i1.2 ◽

2019 ◽

pp. 7-12

Author(s):

Alok Pratap Singh ◽

Iti Chauhan ◽

Snigdha Bhardwaj ◽

Praveen Gaur ◽

S Sadish Kumar ◽

...

Keyword(s):

Method Development ◽

Limit Of Detection ◽

Hplc Method ◽

Uv Detector ◽

Limit Of Quantification ◽

Relative Standard ◽

Validation Parameters ◽

Wide Range ◽

Tract Infections ◽

Development And Validation

Introduction: Azithro-mycin a semi-synthetic, azalide congener of erythro-mycin indicated in the treatment of respiratory tract infections. Various methods available for determination of Azithro-mycin, but HPLC are most versatile one. Objective: The present study is based on the development and validation of a rapid, simple high performance liquid chromatography (HPLC) method equipped with UV detector for quantitative analysis of Azithro-mycin (AZN) in suspension. Material and methods: The Method was performed by using Hypersil BDS-C18 (250 mm × 4.6 mm i.d.) column MS-II, with an isocratic mobile phase of methanol, acetonitrile and phosphate buffer pH 8 (60:30:10; v/v) with run time 15 minutes. The determinations were performed at a flow rate of 1.0ml/min, and UV detector set at 212 nm. Result and Discussion: The method was found to be specific with relative standard deviation (RSD) less than 2.09%. The method showed accuracy with RSD less than 1.34% and precision in repeatability with RSD less than 1.42%. The method was found to be linear over a wide range of concentration from 1.0 to 50.0 μg/mL (R2 = .995). Limit of detection and limit of quantification were found to be 14.40 ng/mL and 43.66 ng/mL respectively. Conclusion: It was advantageous to use UV detector over other methods employing electrochemical, photodiode array etc. as the detector, because of cheap and easy availability. The developed method fulfilled all validation parameters as per ICH and can be successfully applied to quantify percent drug content in marketed oral Azithro-mycin suspension.

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Method Development and Validation for the Simultaneous Determination of Four Coumarins in Saussurea superba by Capillary Zone Electrophoresis

Journal of AOAC International ◽

10.1093/jaoac/93.5.1410 ◽

2010 ◽

Vol 93 (5) ◽

pp. 1410-1415 ◽

Cited By ~ 2

Author(s):

Juan Chen ◽

Li Chen ◽

Hong-Li Zhang ◽

Yan-Ping Shi

Keyword(s):

Simultaneous Determination ◽

Applied Voltage ◽

Method Development ◽

Migration Behavior ◽

Electrophoretic Method ◽

Wide Range ◽

Method Development And Validation ◽

Capillary Zone ◽

Development And Validation

Abstract A capillary zone electrophoretic method has been developed for the determination of four coumarinsskimmin, scopolin, scopoletin, and umbelliferonein Saussurea superba with UV detection at 254 nm. The capillary temperature was kept constant at 25°C. Effects of buffer pH, electrolyte concentration, organic modifier, and applied voltage on migration behavior were studied systematically. The optimum conditions for separation were achieved by using 30 mM borate buffer at pH 9.02 containing 15% (v/v) methanol as the electrolyte and 25 kV as the applied voltage. For all analytes a good linear regression relationship (r >0.999) was obtained between peak area and concentration over a relatively wide range. The method was validated for repeatability, precision, and accuracy. The validated method was successfully applied to the simultaneous determination of the four analytes in S. superba.

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Method Development and Validation for Estimation of related Substances in Tilorone Dihydrochloride using RP¬-HPLC

Research Journal of Pharmacy and Technology ◽

10.52711/0974-360x.2021.00577 ◽

2021 ◽

pp. 3319-3324

Author(s):

M. Zeba Baktiyar ◽

B. Mohammed Ishaq ◽

Siva Sanker Reddy L ◽

Sreenivasulu M.

Keyword(s):

Mobile Phase ◽

Method Development ◽

Hplc Method ◽

Column Temperature ◽

Related Substances ◽

Rp Hplc ◽

Ich Guidelines ◽

Wide Range ◽

Method Development And Validation ◽

Development And Validation

A simple, precise and reproducible RP-HPLC method was developed for the estimation of related substances in tilorone dihydrochloride. Quantification was performed using a Zorbax SB-phenyl column (150 × 4.6mm, 5µ) with mobile phase A: 20mM potassium dihydro phosphate + 2ml of triethylamine, pH 2.30 and mobile phase B: acetonitrile, methanol and water 60: 20: 20% v/v. A gradient program was followed with a run time of 55 minutes at a flow rate of 1.0 ml/min. The column temperature was maintained at 40°C, the injection volume was 10 µl and the detection was performed at 269nm using a PDA detector. The retention time of Tilorone dihydrochloride was found to be 10.36 minutes. The proposed method has been validated according to the ICH guidelines for Linearity, Precision, Accuracy, LOD, and LOQ. The method was linear from 0.157 - 3.934μg/ml for standard, 0.153-3.820μg/ml and 0.166 - 4.140μg/ml for impurities, TLHC01 and TLHC02 respectively. The impurities TLHC01 and TLHC02 have been mapped in all stress conditions. The LOD and LOQ of TLHC01 were found at 1.757μg/ml and 5.857μg/ml and 1.919μg/ml and 6.396μg/ml respectively for TLHC02 respectively. Statistical analysis showed that the method was precision, reproducible, selective, specific and accurate for the analysis of Tilorone dihydrochloride and its impurities. The wide range of linearity, sensitivity, precision, short retention times and simple mobile phase have shown that the method is suitable for the routine quantification of mass impurities of tilorone hydrochloride and its dosage pharmaceutical forms with high precision and accuracy.

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