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Related Concept Videos

Insulin: Biosynthesis, Chemistry, and Preparation01:25

Insulin: Biosynthesis, Chemistry, and Preparation

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The endoplasmic reticulum (ER) of pancreatic β-cells synthesizes preproinsulin, which consists of a signal peptide, A and B chains, and a C-peptide. Preproinsulin is then cleaved and folded into proinsulin, which translocates to the Golgi apparatus for sorting and packaging into secretory granules. In these granules, enzymatic clipping generates insulin and C-peptide.
Damage or functional impairment of β-cells inhibits insulin production, leading to diabetes. Diabetes treatment...
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Insulin Formulations: Types and Delivery01:27

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Insulin preparations are categorized by their duration of action into short-acting and long-acting types. Two strategies are used to modify insulin's absorption and pharmacokinetic profile: slowing the absorption post-subcutaneous injection, or altering human insulin's amino acid sequence or protein structure. These changes retain the insulin's ability to bind to the insulin receptor, but alter its behavior in solution or after injection.
Short-acting insulins are divided into...
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Data Validation01:15

Data Validation

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Method validation is a crucial process in analytical chemistry designed to confirm that a given method consistently produces reliable and high-quality results. This process is essential when a method is applied to different sample matrices or when procedural modifications are made, ensuring that the results meet acceptable standards across various applications.
Key parameters for method validation include:
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Insulin: Dosing Regimen and Adverse Effects01:16

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Insulin-replacement therapy usually includes both long-acting insulin (basal) and short-acting insulin (to cater to postprandial needs). In a diverse group of type 1 diabetes patients, the average daily insulin dose is typically 0.5-0.7 units/kg body weight. However, obese patients and pubertal adolescents may need more due to insulin resistance.
The basal dose constitutes about 40%-50% of the total daily dose, with the rest as premeal insulin. The mealtime insulin dose should mirror...
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Less is more: Validating a single method for comprehensive rh-insulin analysis.

Sanjay Mendiratta1, Gurminder Bindra1, Sukhwinder Singh1

  • 1National Institute of Biologicals, Ministry of Health and Family welfare, Noida 201309, India.

Journal of Pharmaceutical and Biomedical Analysis
|March 28, 2024
PubMed
Summary
This summary is machine-generated.

A new single reverse-phase high-performance liquid chromatography (RP-HPLC) method accurately analyzes human insulin potency and related proteins. This streamlined approach reduces analysis time and cost for quality control.

Keywords:
InsulinMethod ValidationMethod VerificationPotencyQuality ControlRelated Proteins (Impurities)

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Area of Science:

  • Analytical Chemistry
  • Pharmaceutical Analysis

Background:

  • Current methods for human insulin analysis in the Indian Pharmacopoeia involve separate procedures for potency and related proteins.
  • This necessitates multiple analytical runs, increasing time and resource expenditure for quality control.

Purpose of the Study:

  • To develop and validate a single, unified method for the simultaneous analysis of human insulin potency and related proteins.
  • To establish a more efficient and cost-effective quality control process for insulin formulations.

Main Methods:

  • Utilized reverse-phase high-performance liquid chromatography (RP-HPLC) with a C-18 stationary phase.
  • Employed a mobile phase of 55% buffer (0.2 M sodium sulfate, pH 2.3) and 45% acetonitrile, with UV detection at 214 nm.
  • Validated the method for linearity, accuracy, and limit of detection.

Main Results:

  • The method demonstrated excellent linearity over a concentration range of 0.08-4.5 mg/mL (r²=0.999).
  • Achieved a limit of detection of 0.094 mg/mL and accuracy ranging from 99% to 102.8%.
  • Successfully quantitated both potency and related proteins in a single chromatographic run.

Conclusions:

  • A single RP-HPLC method provides a precise and efficient means for analyzing human insulin potency and related proteins simultaneously.
  • This unified approach offers significant advantages in terms of time and cost savings for quality analysis in manufacturing and regulatory settings.
  • Implementation of this method can enhance the availability of standardized, high-quality insulin preparations for public health.