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Nov08,2022
The number one priority in the fine chemicals industry is health, safety, and environment (HSE). Still, it is essential to remember that safety has many levels in fine chemical production. First off is the security of the manufacturing method and the workers. Then there is the security of people living in the vicinity of the works. Finally, there is the security of the end creation itself, which could be, for example, active pharmaceutical ingredients that will be utilized to manufacture drugs. Plant safety systems are employed to track critical parameters to ensure the overall process's safety. If a deviation is noticed that would cause a dangerous situation, changes are automatically executed to restore the plant to a safe condition, disrupting the display. In addition, other parameters that affect product quality also need to be carefully monitored to ensure the end product's safety.

Chemical Sourcing - Manufacturing of only fine chemicals

At first sight, providing chemicals to a customer's supply chain is not new. It is not worth spending much attention to, being the bread-and-butter company of the chemical industry since its emergence. There are thousands of chemical providers worldwide: large and small, locally active and international, young and adult, application-oriented and compound-focused, technology experts, pure manufacturers, traders, refiners, hybrids, and organizations working on the request, while others manufacture for the complimentary market. All the instruments part of this safety process must be accurately calibrated to ensure they are working correctly. Safety systems must also be tested regularly to guarantee they are functioning correctly. This is where automatic calibration answers can help. The Windson Chemical Private Limited solution breaks the process down into four simple steps:
  • Step 1: A work demand is created in the care management system and automatically sent to the calibration software to determine the appropriate calibration methods.
  • Step 2: The device details and calibration procedures are then sent to a documenting calibrator or pill, and the calibration is completed.
  • Step 3: The device automatically records and stores the results, and the user signs off on them electronically.
  • Step 4: The effects are automatically assigned to the calibration software for the warehouse, and the work demand is closed in the maintenance management plan. This provides a complete and easily searchable digital record of each transmitter being calibrated, the person who performed the calibration, and a fully traceable audit trail, including a digital signature and calibration approval.
The Windson Chemical Private Limited system also enables you to include checklists as part of the calibration procedure, which is very important for the safety systems used in fine-chemical production facilities. For example, in ATEX (explosive atmosphere) environments, there are apparent procedures that must be followed to ensure safety. But technology alone is insufficient to ensure safety – industry knowledge is also needed to ensure that all relevant factors are considered. Because Windson Chemical Private Limited has extensive expertise and a long history of working with customers in the fine chemicals industry, we are well-placed to recommend the right solutions, processes, and techniques to help you work safely and effectively in your works with trustworthy calibration that meets your requirements, now and in the future.

Windson Chemical Private Limited Manufacturing Process for fine & speciality chemicals

Our manufacturing processes genesis guarantees our thorough knowledge of the target molecule and its synthesis. This, in turn, lowers the quality risk in the show. Even in cases where the client provides a synthesis method, we never now apply it to make the compound in question to evade both sides facing issues and substantial follow-up costs that can be bypassed if our typical approach is followed: While this approach might seem complex at first sight, it was proven efficient, workable, robust, and advantageous over many years.
Oct08,2022

Introduction

Mutagenic impurities, formerly known as genotoxic impurities (GI) or potential genotoxic impurities (PGI), are compounds that can modify DNA and, consequently, can cause cancer. Impurities present in the marketed drug must be evaluated early in the drug development process. To that end, analytical techniques must be developed that are sharp and specific sufficiently to select the levels in both drug import and product. The International Conference on Harmonization published ICH M7 guidelines, which highlight the requirements for assessing and controlling DNA-reactive impurities to ensure the safety of pharmaceutical products. The European Medicine Evaluation Agency (EMEA), U.S. FDA, and the Asia regulatory agencies follow these guidelines. They require that any mutagenic impurities in a drug substance or product must be below the Threshold of Toxicological Concern (TTC) of 1.5 µg per day based on the Maximum everyday dosage of the pharmaceutical combination over a lifetime. For example, for a dosage of 1 g of Active Pharmaceutical Ingredient (API) per daytime, any impurity must be smaller than 1.5 ppm (1.5 µg). This is orders of magnitude lower than general pharmaceutical impurities analysis, which is at the 500-ppm level and governed by Q3B(R).

What is Methyl 3-Aminocrotonate?

Methyl-3-aminocrotonate (MAC) is a Michael-reactive receptor and a starting material in different cardiovascular drug products. The API used is an active sense from a proprietary pharmaceutical product; therefore, just the partial structure is shown in Figure 1. MAC flags up a favorable from the mutagenic structural attention. This compound would typically be analyzed by static head space (SHS) GC-MS after derivatization with trifluoroacetic anhydride to increase the volatility (Figure 2).3 When MAC (underivatized) was analyzed by UPLC-MS, nothing was noticed; this was supposed to be due to its insufficient stability in aqueous solvents.     In this type of trace analysis, where there is a large amount of matrix, it would be advantageous if chemical derivatization of the mutagenic impurity could be avoided for the following reasons: The shape of acylation derivatives can be challenging to prepare Response bi-products can happen, which could add more complication to the matrix The different derivatization step would need extra validation to be completed

Benidipine

Benidipine is a manufactured dihydropyridine calcium channel blocker used to dine hypertension and angina pectoris.

Indication

Benidipine is a powerful and long-lasting medicine for treating cardiovascular conditions such as hypertension, renoparenchymal hypertension and angina pectoris.

Pharmacodynamics

Benidipine reduces systolic and diastolic blood pressure and presents a decrease in heart rate pulse after treatment. It is also reported a decrease in urinary protein excretion and serum triglycerides. Different studies have shown benidipine anti-oxidative action, stimulation of NO production, suppression of bonding molecules presentation, stimulation of osteoblast differentiation, and suppression of the increase of vascular soft muscle cells and mesangial cells as myocardial saving. The enhancement of NO production is associated with benidipine's cardioprotective and antiatherosclerotic effects.

Absorption

Benidipine is rapidly spread in oral administration, reaching the highest concentration within 2 hours. The short time needed for maximum concentration to reach a particular characteristic of benidipine compared with other calcium channel blockers. The registered maximum concentration and AUC are dose-dependent, and they can go from 0.55-3.89 ng/ml and 1.04-6.7 ng.h/ml, respectively, when administered at a dose of 2-8 mg.

Volume of distribution

Benidipine is highly distributed to the tissues mostly in the liver, kidneys, and plasma. It does not offer a high expansion following repeated oral administrations.

Side Effects of BENIDIPINE

  • Fatigue
  • Ankle swelling
  • Sleepiness
  • Flushing (sense of warmness in the front, ears, neck and chest)
  • Headache
  • Dizziness
  • Palpitations
  • Nausea
  • Oedema (swelling)
  • Abdominal pain

Cilnidipine

Cilnidipine is a dihydropyridine calcium channel blocker with activity on both N- and L-type calcium media used to treat hypertension.

Indication

Cilnidipine is suggested for the management of hypertension for end-organ security. It is reported to be useful in elderly patients and those with diabetes and albuminuria. Cilnidipine has been increasingly utilized in patients with chronic kidney disease. Hypertension is the phrase used to describe the existence of high blood pressure. Blood stress is generated by the power of the blood pumped from the heart against the blood vessels. Thus, hypertension is generated when too much pressure is on the blood vessels, damaging the blood vessel.

Pharmacodynamics

The administration of cilnidipine has been shown to present an antisympathetic profile in vitro and in vivo. It decreases blood pressure carefully and effectively without excessive blood pressure decrease or tachycardia.

Absorption

Cilnidipine offers an extremely rapid absorption with a highest peaked concentration after 2 hours. Its distribution tends to be higher in the liver and kidneys, plasma and other tissues. Cilnidipine does not offer a high accumulation in the tissue following repeated oral administration. Cilnidipine is reported to offer very low bioavailability, approximately 13%. This low bioavailability is suggested due to its low aqueous solubility and high permeability. Hence, efforts have been made to find an innovative formulation that can significantly improve the bioavailability of this drug. One of these formulations compares the generation of polymeric nanoparticles, which improves the bioavailability by 2.5-3-fold.

Volume of distribution

Drugs in the group of dihydropyridines, such as cilnidipine, tend to have a large volume of distribution.

Side Effects of CILNIDIPINE

  • Headache
  • Feeling exhausted
  • Swollen ankles
  • Nausea

Isradipine

Isradipine is a dihydropyridine calcium drain blocker utilized for the therapy of hypertension.

Pharmacodynamics

Isradipine decreases arterial soft muscle contractility and subsequent vasoconstriction by impeding the influx of calcium ions through L-type calcium media. Calcium ions joining the cell through these channels bind to calmodulin. Calcium-bound calmodulin then attaches to and activates myosin light chain kinase (MLCK). Activated MLCK catalyzes the phosphorylation of myosin's regulatory light chain subunit, a key phase in muscle contraction. Signal amplification is performed by calcium-induced calcium departure from the sarcoplasmic reticulum through ryanodine receptors. Inhibition of the initial inflow of calcium decreases the contractile action of arterial soft muscle cells and results in vasodilation. The vasodilatory results of isradipine result in a general decrease in blood pressure.

Absorption

Isradipine is 90%-95% absorbed and is subject to extensive first-pass metabolism, resulting in a bioavailability of about 15%-24%.

Toxicity

Symptoms of overdose include lethargy, sinus tachycardia, and temporary hypotension. Significant lethality was followed in mice presented with oral doses of around 200 mg/kg, and rabbits provided about 50 mg/kg of isradipine. Rats tolerated amounts of over 2000 mg/kg without consequence in survival.
Sep15,2022
Active Pharmaceutical Ingredients are the varied elements of a medical drug. These drugs are produced to Supply the desired impact on the human body which helps in treating Many medical conditions. Multiple app chemical combination mixtures are processed to make an API that is added to the drugs. The first material in a particular biologic medicine is the BPI or Bulk Process Intermediate. What are APIs? People take medicines to remedy, diagnose, treat, or detain a disease. The majority of medications have a few components. The most vital one is the API, which stands for Active Pharmaceutical Ingredient. This is the real substance in the drug that has to do the work in your body. For instance, the API in Aspirin is “acetylsalicylic acid“. This API will try to fight the molest of your fever or headache. Some Drugs carry the name of the API. For Example, the API of Paracetamol is “paracetamol“. To make a pill, you’ll require more ingredients than the API. That’s where excipients come in. These ingredients are used to issue the medicine, for instance, volume, a sweet flavor, or color. API is not purely used in the pharmaceutical industry, but in the web developers group, API is General. For them, though, it stands for “Application Programming Interface“. Elements of Medicines Every medicine is made up of 2 core elements, i.e., the API is the Primary ingredient and the excipient, which is the material other than drugs that support delivering the drugs to the system. The excipients are chemically passive compounds in the tablet, like mineral oil or lactose. Therefore, the Active Pharmaceutical component isn’t made by just 1 Cross Action from raw matters; instead, it becomes the API through many chemical compounds. Types of Pharmaceutical Drug- Active Pharmaceutical Ingredients API market is segmented into two varied, i.e., Synthetic and Natural. We have taken an Equal concept in their manufacturing department. Each medication is a Crossbreed of the two varieties. Generic and novel are other categories for synthetic APIs. Finally, according to their solvency, APIs are often categorized as soluble or insoluble. Synthetic Active Pharmaceutical Ingredient Synthetic chemical combinations are also called small molecules. it represents a considerable part of the pharmaceutical Bazar. A number of small molecular medications that are Sold commercially assist in this. Natural Medicine The making of biological drugs uses the original form of API. These medicines are replacing the top-selling products in the Field of medicine. Even though uprising demands, biological drugs are less common than small molecule drugs. Quality of APIs (Active Pharmaceutical Ingredient) Medication is, of course, intended to aid people. So, we require that the ingredients, such as the API, are Secure to use. No matter where an API is produced, it should fall on the safety and quality criteria of the country where end users are located. That means, Medicines sold in the E.U. need to meet the strong safety and quality standards of the European Medicines Agency, and those sold in the U.S. need to meet the regulations given by the U.S. Food and Drug Administration (FDA). Companies in the entire API supply chain get inspected by their government. Foreign government instances could also examine them, and third-party companies check each other. When Each criterion is up to code, companies issue a certain certificate, such as GMP or a written Support, so other professionals in the production know that the Definitive company complies with the manufacturing standards without having to survey or audit themselves. When the inspection fails, the companies will be given a warning, and pharmaceutical buyers won’t be competent to buy there until the matter is resolved and the company resurveyed. Not only the plant or production Convenience is inspected, but laboratories also analyze every Group of produced API; depending on the situation, it is also possible that one Correct batch is analyzed multiple times: For instance, by the Business that created it, by a 3rd party laboratory, by the businessman who buys it, and by the medical center that will use it. The difference between Q.A. and Q.C. Also, every pharmaceutical business has an entire Batch dedicated to quality certainty and quality control. This team contains qualified persons such as pharmacists, chemists, biologists and other trained personnel. The first team, recognize assurance, ensures everything is as per standards before the production even starts, while the recognize control checks ensure that everything was performed as per planning.
Jul21,2022
The pharmaceuticals used as raw materials for the creation of bulk drugs are known as bulk drug intermediates. They can also refer to a substance created during the synthesis of an API that needs to go through additional molecular processing or alteration before it is an API. Drug intermediates are produced hygienically from premium raw ingredients for the pharmaceutical and cosmetic industries. Drug intermediates are used in research and development by pharmaceutical businesses. Various forms of drug intermediates include pharmaceutical intermediates, veterinary intermediates, and bulk intermediates. A high rate of growth in the global market for medicinal intermediates is anticipated due to increased global R&D activity. The market for drug intermediates is divided by regions, end users, and intermediate types. The market is divided into two primary types based on the type of intermediate: The raw substances or unprocessed medications that constitute the basis for therapeutic activity are known as API intermediates, or active pharmaceutical ingredients. These are the active ingredients that are subsequently transformed into a variety of forms, including tablets, capsules, solutions, etc. Even API functions as a drug. Advanced types of bulk drug intermediates are referred to as advanced intermediates. In order to carry out drug interaction activity created in novel chemical products, advanced intermediate pharmaceuticals are used. The pharmaceutical, biotechnology, and chemical industries are the three subsectors under which the global market for drug intermediates is divided according to end user. There are several different types of drug intermediates, including low quality intermediates, high quality intermediates, and premium quality intermediates. The high- and premium-quality medication intermediates are primarily utilised for academic research. Since the biotechnology and life sciences industries are expanding quickly, along with rising adoption and expanding application in research disciplines, there has been an ever-increasing global demand for high-quality medication intermediates. Bulk Drug Intermediates Pharmaceutical, biotechnology, and research businesses are putting more of an emphasis on R & D in drug discovery and development. The drug intermediates market is divided into five regions based on geography: North America, Europe, Asia Pacific, Latin America, and the Middle East & Africa. Due to the region's expanding pharmaceutical and life sciences industries, particularly in emerging economies like India and China, Asia Pacific now dominates the market for drug intermediates. Due to the advancement and expansion of increasingly effective and cutting-edge technology, Europe is now the second-largest market for medication intermediates. Because of rising healthcare spending, the adoption of western lifestyles, and expansion in research and development, it is also projected that the Asia Pacific drug intermediate market will rise at a rapid rate. The global market for drug intermediates is anticipated to rise as a result of increased competition in the pharmaceutical and biotechnology sectors for outsourcing the production of medication intermediates. One of India's top producers of pharmaceutical intermediates is WindSon Chem. We offer carbonyl compounds with CAS numbers like 4023-34-1 and 2719-27-9. Due to our innovations for chemical reactions, we have cyclohexanecarbonyl chloride (CAS No. 5500-21-0) and cyclopropanecarbonitrile in our portfolio. Bulk Drug Intermediates Tags - DBA Intermediates of Lumefantrine | Manufacturers | Suppliers | India | Bulk Drug Intermediates
Jun14,2022
DBA providing in Gujarat - Given the shifting outlook of Chinese manufacturing units, pharmaceutical intermediates businesses are among the most active industries in India today. Considering the needs of the Indian API business, which is heavily reliant on Chinese suppliers, this unit might be developed to meet those needs. pharmaceutical firms in India To begin, we will produce 2, 7-Dichloro-alpha-[(dibutylamino)methyl]-9H-fluorene-4-methanol (DBA) is an anti-malarial medication intermediate named Lumefantrine. DBA providing in Gujarat 1) DBA:  2-(2, 7-DICHLORO-9H-FLUORENYL-4-YL)OXIRANE (Oxirane) is reacted with Di-nbutyl amine in presence of a catalyst & n-Butanol as reaction medium. The solids formed in the reaction are filtered & dried under vacuum to get the product. Type of reaction: Epoxide opening with secondary amine. Chemical reactions :  2-(2,7-Dichloro-9H- Fluorenyl-4-yl) Oxirane Mol wt: 277.15 Di-n-butylamine Mol wt: 129 DBA Mol wt : 406.39 DBA providing in Gujarat 2) Tert-Butyl [(1S, 2R)-1-benzyl-2-hydroxy-3-(isobutyl amino) propyl]carbamate:  (2S,3S)-1,2-Epoxy-3-(Boc-amino)-4-phenylbutane (epoxide) is reacted with iso-butyl amine to get the product. The product is isolated in presence of Toluene+Methanol. Type of reaction: Epoxide opening with primary amine. Chemical reaction: (2S,3S)-1-2-epoxy-3-(boc-amino)-4-phenylbutane Mol wt: 263-34 Isobutyl amine Mol wt: 73 Tert-Butyl-[(1S,2R)-1-benzyl-2-hydroxy-3-(isobutylamino)propyl] carbamate Mol Wt: 336.47 DBA providing in Gujarat 3) 4-(2-Aminoethyl) phenol:  L-Tyrosine is heated in presence of a catalyst using N-methyl pyrrolidone/Diphenyl ether as reaction medium. After the completion of reaction the reaction medium is distilled out under vacuum. Toluene+Methanol is added to concentrated mass to get the product, which is filtered & dried. Type of reaction: Decarboxylation. Chemical reaction:  L-Tyrosine Mol wt: 181 4-(2-Aminoethyl)phenol Mol wt: 137 DBA providing in Gujarat 4) Methyl 2-(1,8-diethyl-1,3,4,9-tetrahydropyrano[3,4-b]indol-1-yl)acetate:  7-Ethyl tryptophol(7-ETP) is reacted with Methyl -3-oxo pentanaote in presence of a catalyst & using MDC as reaction medium. Once the reaction is over the catalyst is filtered (recycle in next batch) & MDC is distilled out. Methanol is added to isolate the product. Solids are filtered & dried to get the product. Type of reaction: Six membered ring formation. Chemical reaction:  7-Ethyl tryptophol Mol wt: 189.25 Methyl-3-pentanoate Mol wt: 130 Methyl-291,8-diethyl-1,3,4,9-tetrahydropyrano[3,4]-indol-1-yl acetate Mol wt: 301.35 DBA providing in Gujarat 5) Tetra methyl-1,4,7,10-Tetraazacyclododecane-1,4,7,10-tetraacetate:  1,4,7,10-tetrazacyclododecane (cyclen) is reacted with Methyl bromo acetate in presence of a catalyst, Potassium carbonate & using Acetonitrile as reaction medium. After the completion of reaction, salt formed & excess potassium carbonate is filtered. The filtrate is distilled to recover Acetonitrile & get the product. Type of reaction: N-alkylation reaction Chemical reaction: Cyclen Mol wt: 172 Methyl bromoacetate Mol wt: 153 Dota ester Mol wt: 460 DBA providing in Gujarat Tags - DBA intermediates of Lumefantrine | DBA providing in Gujarat | 2,7-Dichloro-alpha-[(dibutylamino)methyl]-9H-fluorene-4-methanol in Gujarat | Intermediates of Lumefantrine in Gujarat | 2,7-Dichloro-alpha-[(dibutylamino)methyl]-9H-fluorene-4-methanol manufacturer | Intermediates of Lumefantrine manufacturer | DBA manufacturer
Jun07,2022
The Chemical Abstracts Service Registry assigns a CAS number, which stands for Chemical Abstracts Service number, to every chemical compound that has existed since the early 1900. It is affiliated with the American Chemical Society, and the numbers assigned to compounds correspond to the order in which new chemicals are discovered. The significance of the number about the chemical A compound's numerical representation is limited to ten digits, with hyphens used to divide it into three pieces. The right-most digit, known as the check digit, is used to validate the number's validity and uniqueness. The CAS number is unique to a compound and has no bearing on the chemical formula or the substance itself. What is the value of the CAS number? The importance of knowing your CAS number is that it allows you to avoid confusion when recognizing compounds and gives you access to a wealth of information. It is the most dependable platform in use around the world, and it acts as a link between numerous nomenclature phrases that describe substances. Validation is simple and reliable. An illustration of the applicability of CAS No. Ethanol is a highly common chemical that goes by a variety of names. Absolute booze Alcohol The spirit of Cologne Alcohol derived from ethyl hydrate of ethyl Hydroxide of ethyl Ethylol Its formula can be written as C23H29Cl2NO, C2H6O, CH3CH2OH, or C2H5OH, among others. With so many distinct ways to refer to the same substance, CAS No. 69759-61-1, 64-17-5, and easy identification can help to clear up the confusion. Conclusion Although the stated example is simplistic, recognizing stereoisomers with a CAS number is a godsend and the quickest and most reliable approach to identifying each chemical. TAGS DBA providing in Gujarat | 2,7-Dichloro-alpha-[(dibutylamino)methyl]-9H-fluorene-4-methanol in Gujarat | Intermediates of Lumefantrine in Gujarat | 2,7-Dichloro-alpha-[(dibutylamino)methyl]-9H-fluorene-4-methanol manufacturer in gujarat | 2,7-Dichloro-alpha-[(dibutylamino)methyl]-9H-fluorene-4-methanol manufacturer | Intermediates of Lumefantrine manufacturer | Lumefantrine manufacturer | cas no 69759-61-1 | CAS 69759-61-1 | dahej | ankleshwar | gujarat | india
May27,2022
DBA Intermediates of Lumefantrine | Manufacturers | Suppliers | India The list of Lumefantrine intermediates that we offer can be found below. Please click on the intermediate name to see additional information about the intermediate you desire. Let's say you can't find the Lumefantrine intermediate you need on this website. In such case, email us a sourcing request for the Lumefantrine intermediate you're seeking for, along with its Cas No and Intermediate name, and we'll get back to you with further information as soon as possible. To send an email, please click here. Please feel free to contact us if you require any upstream or downstream intermediates for your Intermediates of Lumefantrine requirement. We will find the right manufacturer, supplier, or principals according on your specific needs, who are dependable and match your quality standards. INTRODUCTION Primaquine (PQ) diphosphate is a prototype antimalarial medication in the 8-aminoquinoline class. It is the only medicine on the market that prevents relapses of infections caused by the reactivation of dormant forms of the malaria parasites Plasmodium vivax or Plasmodium ovale (also known as hypnozoites) in the liver. PQ is also effective against all developing malaria parasites in the liver and mature P. falciparum gametocytes in the blood. Despite its poor action against asexual blood stages of P. falciparum and other malaria parasites, PQ is the only medicine capable of substantially treating P. ovale and P. malariae when taken in combination with a schizonticide agent. Infections with vivax Despite its unique role in the physician's arsenal for treating malaria, PQ's usage in clinical practise is restricted since it (together with pamaquine, tafenoquine, and a few other medications) can induce severe hemolytic anaemia in people with glucose-6-phosphate dehydrogenase deficiency. PQ can also cause methemoglobinemia, but this toxicity seldom causes noticeable symptoms and is usually self-limiting. Furthermore, the therapeutic response to PQ is dosage and drug-metabolizing enzyme phenotype dependent. PQ is a pro-drug transformed by CYP2D6 enzymes into oxidised metabolites responsible for activity against dormant forms of P. ovale and P. vivax in the liver, according to investigations conducted over the last ten years. In human populations, the CYP2D6 enzyme is highly polymorphic, and patients with a CYP2D6 poor metabolizer phenotype do not react to PQ therapy. To cure malaria, the PQ dose must also be changed in overweight people. As a result, sensitive and reliable methods for analysing PQ in blood plasma are required for monitoring medication levels in patients receiving therapy and studying the kinetics of this antimalarial agent in animal models. DBA Intermediates Table 1 highlights the key features of HPLC techniques for the determination of PQ in biological matrices (such as whole blood, serum, plasma, liver tissues, and blood cells) that have been described to date. The majority of PQ analysis methods in plasma use a liquid-liquid extraction approach followed by protein precipitation (PP), although some also use solid phase extraction (SPE) or a mix of liquid-liquid extraction and SPE procedures. When PP and SPE procedures are used, PQ recovery rates are often higher. The mobile phases varies among available methods, as indicated in. Organic modifiers are frequently added to a mixture of organic solvents (methanol or acetonitrile) with water (acid/basic or neutral) or a buffer solution. The most frequent mode of elution is isocratic, however some procedures use the more time-consuming gradient mode. Most methods use reversed phase C18 columns with various lengths, internal diameters, and particle sizes as the stationary phase, while a few others use silica-modified stationary phases like phenyl and cyanopropyl. All approaches necessitate large sample quantities (500L of plasma), which limits their application in investigations with small rodents like mice and hamsters. PQ's long retention durations mean that these chromatographic procedures will take a long time to analyse (8 minutes). Furthermore, a DAD-UV detector has high detection and quantification limits. We designed and validated a rapid, sensitive, cost-effective, and robust HPLC-DAD-UV approach for analysis of PQ in blood plasma due to the limitations of existing methods. Because this approach necessitates small plasma volumes, it is particularly well suited to studies of PQ kinetics in small rodents. METHODS Reagents All of the compounds were analytical reagent grade or purer. Tedia (Rio de Janeiro, Brazil) provided HPLC-grade methanol and acetonitrile, and Lichrosolv ultrapure acetonitrile was used to test robustness (CAS 75-05-8; Merck Millipore, Darmstadt, Germany). Merck Millipore provided ammonium acetate (CAS 631-61-8, CH3COONH4), zinc sulphate heptahydrate (ZnSO4•7H2O), and acetic acid (CAS 64-19-7, CH3COOH). Genix Indstria Farmacêutica Ltda (PP3016PQRJ, 98-102 percent; Anápolis, Brazil) provided the PQ diphosphate standard. Hipolabor provided sodium heparin (5,000IU/mL-1) for this study (Belo Horizonte, Brazil). A Milli-Q® purifying water system delivered ultrapure water on a daily basis (Merck-Millipore). DBA Intermediates Solutions PQ diphosphate aqueous solutions were made by dissolving 17.6mg of the salt in ultrapure water to a final volume of 50mL, equating to 200g.mL-1 PQ base. Stock solutions were held at -20°C in amber flasks. PQ is stable for at least 7 days in this storage circumstances (as determined by the signal area of a standard solution in an HPLC-DAD-UV analysis). The working solutions were made via repeated dilution of the stock solution on a regular basis. 0.77g ammonium acetate was dissolved in 1L ultrapure water to make ammonium acetate buffer (10mM). A 10% w/v acetic acid solution was used to modify the pH of the solution to 3.8. [DBA Intermediates] Animals Blood plasma samples from female DBA-2 mice were used to perform quantitative PQ tests utilising the novel approach. The Oswaldo Cruz Foundation Central Animal House (CECAL-FIOCRUZ, Rio de Janeiro, Brazil) provided the animals, who were 7 weeks old when they arrived at the laboratory. All of the mice were kept in normal mouse plastic cages with stainless steel lids and bedding made of white Pinus wood shavings (5 mice per cage). Animal cages were kept in a controlled environment (22°C, about 70% relative humidity, and a 12-hour light/dark cycle with lights on at 8 a.m.) and were given free access to filtered tap water and commercially available pelleted food for rats and mice (Nuvital; Nuvilab, Curitiba, PR, Brazil). DBA Intermediates Concerns about ethics The FIOCRUZ Ethics Committee on Animal Use accepted the experimental procedure (2012, P-84/10-7). Treatment Mice (n = 6) were given freshly produced PQ solutions in ultrapure water (2mg PQ base•mL-1 = 3.551mg PQ diphosphate•mL-1). PQ concentrations are measured as PQ base per milliliter-1. PQ aqueous solutions were stored refrigerated (4°C) and light-protected until used within 24 hours of preparation. Oral gavage was used to give PQ diphosphate dissolved in water at dosages comparable to 20 mg PQ•kg-1. Cardiovascular puncture was used to obtain heparinized mouse blood samples 15, 30, 60, and 90 minutes after PQ administration. [DBA Intermediates] Samples of plasma Centrifugation at 12,000 rcf for 15 minutes separated plasma from heparinized whole blood. Plasma samples were immediately transferred to Eppendorf tubes after separation and stored at -20°C until analyses, which were completed within 7 days of PQ delivery. DBA Intermediates Procedure for extracting PQ PP was utilised, followed by drug extraction from plasma, due to its simplicity and inexpensive cost. The following PP conditions were empirically optimised and standardised. 50L of plasma was transferred to 1.5-mL Eppendorf® vials, which were then filled with 50 L of acetonitrile acidified with 2% acetic acid (w/v). For 30 seconds, the tubes containing plasma and acetonitrile were vortexed carefully, avoiding any contact of the mixture with the tube tip. After that, each tube received 25L of an aqueous 12.5 percent zinc sulphate solution (w/v), which was vortexed for 30 seconds. The suspension was then allowed to stand for 30 minutes to allow the plasma protein to fully precipitate. After centrifuging the tubes at 12,000 rcf for 15 minutes, the supernatant was analysed using HPLC-DAD-UV. [DBA Intermediates] chromatographic conditions and equipment The Shimadzu Class-VP (liquid chromatographer attached to a Shimadzu UV detector with the diode array SPD M10A VP equipped with a SCL 10A VP controller, DGU14A degasser, 10ADVP LC binary pump, CTO 10ASVP oven, and SIL10AF autosampler) was used to perform the analyses. Shimadzu Class VP® software, version 6.1, was used to analyse the chromatograms. As mobile phases, several combinations of acetonitrile, methanol, and ammonium acetate buffer were utilised. Before use, all buffer solutions were filtered via a Merck-Millipore 0.45-m pore PVDF filter. Silica-based C18 (250mm 4.6mm i.d. 5m, ODS Hypersil, Thermo, Waltham, MA, USA) and modified-silica cyanopropyl (250mm 4.6mm i.d. 5m, Supelcosil LC-CN; Supelco, St. Louis, MO, USA) HPLC columns were used. For all analyses, the injection volume was 20L. DBA Intermediates Validation The newly developed method was validated using regulatory guidance documents such as the Brazilian Health Surveillance Agency's (ANVISA) validation manual RE 899/2003, the National Institute of Metrology, Quality and Technology's (Inmetro) guidance on the validation of analytical methods (DOQ-CGCRE-008/ 2007), ICH technical requirements for validation of analytical procedures (1995, 1996), and HPLC analysis manuals and handbooks. Selectivity, linearity, intra-day precision, inter-day precision, accuracy, recovery, robustness, and limits of detection and quantification were the metrics utilised to evaluate analytical performance. Standard PQ solutions in ultrapure water were used to create calibration curves for nine concentrations (13, 30, 50, 100, 250, 500, 1,000, 1,500, and 2,000ng.mL-1). Three calibration curves were collected on three different days to assess linearity. A study using blank mouse plasma and mouse plasma spiked with a PQ solution of 250ng.mL-1 determined selectivity. [DBA Intermediates] Homoscedasticity was determined by comparing experimental and estimated values obtained by fitting a linear model to calibration data. Five distinct PQ concentrations (13, 100, 250, 500, and 2,000ng.mL-1) were measured in duplicate three times over a single day to determine intra-day precision. These five concentrations were tested in duplicate once a day on three different days to assess inter-day accuracy. The accuracy of calibration curves (13, 100, 250, 500, and 2,000ng.mL-1) was determined using the formula accuracy = (EAVR/TC) 100, where EAVR is the experimental average and TC is the theoretical concentration. For spikes of high, midrange, and low PQ concentrations, recovery was determined (250, 500, and 1,000ng.mL-1, respectively). [DBA Intermediates] With seven chromatographic variables: buffer concentration, acetonitrile percentage in the mobile phase, flow rate, mobile phase pH, oven temperature, precipitation duration, and brand of commercially available acetonitrile, robustness was tested using Youden's test. With a PQ concentration of 500ng.mL-1, these seven parameters were mixed in eight random chromatographic trials (nominal condition + seven variants) (corresponding to an intermediate point along the standard analytical curve). Using the equations LOQ = DPa (standard deviation of intercept Y) 10/IC (curve slope) and LOD = DPa 3/IC, the limits of detection (LOD) and quantification (LOQ) were calculated from three calibration curves. Analytical statistics The information is presented as means and standard deviations. One-way ANOVA was used to examine differences between two means, followed by Bonferroni post-hoc testing. When p 0.05, differences were judged statistically significant. GRAPHPAD PRISM6® was used for statistical analysis. DBA Intermediates RESULTS A new approach was developed based on laboratory analytical methodologies and previously published HPLC methods for antimalarial drug analysis. The performance of various acetonitrile or methanol combinations with acetate buffer as the mobile phase and RP-C18 silica or modified-silica cyanopropyl columns as the stationary phase was investigated. Using a modified-silica cyanopropyl column (250mm 4.6mm i.d. 5m, Supelcosil LC-CN) and a 45:55 mixture of acetonitrile and 10mM ammonium acetate buffer (pH = 3.80) as the mobile phase, optimal HPLC conditions for the measurement of PQ in blood plasma were found. The use of organic mobile phase modifiers turned out to be unnecessary. [DBA Intermediates] The flow rate was 1.0 mL.min-1, the temperature was 50 degrees Celsius, and the absorbance (UV) was measured at 264 nanometers. The proposed method for measuring PQ in blood plasma has a quick analysis time (t R = 5.80 0.20 min and a total analysis time of 7 min), strong signal symmetry (1.14), a high retention factor (k = 2.15), and low detection and quantification limitations. Analyses of blank plasma samples and plasma samples spiked with PQ were used to determine the method's selectivity and specificity. A, B, and C depict the PQ signal from the plasma extract at 5.94 min, blank plasma, and overlapping chromatograms, respectively. Around t R 5.9 min, no interference was identified for the PQ chromatographic window, demonstrating the method's selectivity and specificity. For 50-L plasma samples, a method for extracting PQ from the plasma was developed. Acetonitrile was added first, followed by zinc sulphate to achieve full PP (see Methods). The overall time required for complete PP and PQ extraction was around 45 minutes. DBA Intermediates PQ recoveries were 89.8 5.1 percent at 250ng.mL-1, 97.7 1.1 percent at 500ng.mL-1, and 100.4 4.6 percent at 1,000ng.mL-1 from plasma spiked with PQ diphosphate (stated as PQ base). The recovery of 80 percent or greater additional PQ at all tested concentrations20,23 revealed that the extraction procedure designed for small sample volumes (50L) was highly efficient. The linearity, intra-day and inter-day precision, accuracy, recovery, robustness, and detection and quantification limitations of the HPLC-DAD-UV technique for the determination of PQ in plasma matrices developed in this work were all validated (see Methods). Excellent linearity was observed in the calibration curves generated in the concentration range of 13 to 2,000ng.mL-1 (r = 0.9997 0.0003; average and standard-deviation of three different calibration curves). [DBA Intermediates] In the concentration range of 13 to 2,000ng.mL-1, the discrepancy between experimental and calculated values revealed that the distributions of values were homoscedastic. Concentration = [(absorbance) + 185.58 (36.14)]/103.29 (0.67) was the linear regression equation for PQ quantification in plasma samples. The calculated LOQ and LOD were 3.5 and 1.0ng.mL-1, respectively (see Methods). Precision and accuracy are offered both intra-day and inter-day. Values for precision and accuracy were within acceptable limits (85-115 percent ). The method revealed the most variation in PQ content as a function of the mobile phase pH (13.7%) and, to a lesser extent, as a function of the acetonitrile brand (0.9 percent ) DBA Intermediates DISCUSSION HPLC-UV methods for PQ analysis in plasma that have previously been reported require large sample volumes (100-500L) and were developed and validated using human blood plasma. We developed a new extraction approach for PQ analysis in small amounts of plasma, which is required for investigations in small animals (such as DBA-2 mice), the most common experimental model of blood stage malaria. [DBA Intermediates] DBA-2 mice are vulnerable to Plasmodium berghei (ANKA) infection and are commonly employed to research the physiopathology of severe and deadly malaria as well as to test new antimalarial drugs. Furthermore, the extraction technique is straightforward because it does not require solid phase extraction; the overall analysis time is brief (7 minutes); and the mobile phase consists of ultrapure water buffer and acetonitrile, with no organic modifiers required. The method is likely to be more cost-effective than traditional analytical methods for determining PQ in blood plasma. Because a UV detector was utilised, the observed LOQ and LOD (3.5 and 1.0ng.mL-1, respectively) were extremely low. In fact, LOQs for PQ studies utilising HPLC-UV have been reported in the literature as high as 10ng.mL-1. The new method's outstanding signal symmetry and linearity over a wide range of concentrations resulted in significantly lower LOQ and LOD estimations than similar HPLC methods for PQ previously described. [DBA Intermediates] Variation in robustness varied from 0.9 percent (acetonitrile brand) to 13.7 percent (acetonitrile brand) (mobile phase pH). Nonetheless, fluctuation in any of the seven evaluated factors had no significant effect on the analytical assessment of PQ content (variation 15%), suggesting that the procedure is reasonably robust. In triple analyses of female DBA-2 blood plasma, the relative standard deviation was less than 5%, suggesting that drug plasma extraction and analysis are effective and repeatable. It was possible to develop a new HPLC method for the quantification of PQ in small volumes of plasma that is suitable for kinetic studies of this compound in small rodents, including mouse models for the study of malaria, by combining a simple and relatively low-cost extraction procedure with a robust and validated analysis method. Selectivity, linearity, intra-day and inter-day precision, resilience, accuracy, and recovery are all strengths of this innovative approach. A quicker analysis time, the use of smaller amounts of plasma, and lower LOD and LOQs are all advantages of the analytical method over similar, current HPLC-UV procedures. DBA Intermediates Tags - 69759-61-1 | CAS 69759-61-1 | DBA intermediates of Lumefantrine | DBA providing in Gujarat | 2,7-Dichloro-alpha-[(dibutylamino)methyl]-9H-fluorene-4-methanol in Gujarat | Intermediates of Lumefantrine in Gujarat | 2,7-Dichloro-alpha-[(dibutylamino)methyl]-9H-fluorene-4-methanol manufacturer | Intermediates of Lumefantrine manufacturer | DBA manufacturer | Suppliers | India |