Development and Validation of an HPLC Method for Simultaneous Determination of Capsaicinoids and Camphor in Over-the-Counter Medication for Topical Use

Abstract

A reverse-phase high-performance liquid chromatography method was developed to determine and quantify capsaicin (trans-8-methyl-N-vanillyl-6- nonenamid), dihydrocapsaicin (8-methyl-N-vanillylnonanamide), and camphor (trimethylbicyclo[2.2.1]heptan-2-one). It is applicable in analyses of over-the-counter (OTC) medications for topical use and raw materials such as chili pepper oleoresin. Chromatographic separation was carried out on a C18 column using an isocratic flow of the mobile phase containing acetonitrile and ultrapure water in a ratio of 2:3, with pH adjusted to 3.2 using glacial acetic acid, and a flow rate of 1.5 mL/min. The concentration of the eluting compounds was monitored by a diode-array detector at a wavelength of 281 nm. The method was evaluated for several validation parameters, including selectivity, accuracy (confidence intervals < 0.05%), repeatability, and intermediate precision. The limit of detection (LOD) was determined to be 0.070 µg/mL for capsaicin, 0.211 µg/mL for dihydrocapsaicin, and 0.060 µg/mL for camphor. The limit of quantification (LOQ) was determined to be 0.212 µg/mL for capsaicin, 0.640 µg/mL for dihydrocapsaicin, and 0.320 µg/mL for camphor. Linearity was set in the range of 2.5–200 µg/mL for capsaicin and dihydrocapsaicin and 25–2000 µg/mL for camphor. The suggested analytical method can be used for quality control of formulated pharmaceutical products containing capsaicinoids, camphor, and propolis.

High performance liquid chromatography (HPLC) based genetic diversity profiling of chilli germplasm for fruit pungency and phytochemical contents

“…The multivariate analysis method also known as principal component analysis (PCA) is a simple way to extract required information from large set of data and also applied for assessing genetic diversity [ 7 ]. The role of eigen values is to quantify the individual component to the gross variance and each individual eigen vector depicts the percent contribution of a variable to its respective PC [ 8 , 14 ].…”

“…Some minor capsaicinoids are also present e.g., nordihydro-capsaicin (NDC ), homodihydro-capsaicin (HDC) and homocapsaicin (HDC) are half as potent as major capsaicinoids and measured in terms of Scoville heat units (SHU) [ 6 ]. The amount of capsaicinoides is 0.003-1% in total weight of plant in average [ 7 ]. Capsaicinoids comprises 90% of CAP and DhCAP and are divided into branched chain fattyacid and phenylpropanoid pathway [ 8 ].…”