Carotenoid extraction from the gonad of the scallop Nodipecten nodosus (Linnaeus, 1758) (Bivalvia: Pectinidae)

Abstract

In marine bivalve mollusks, unsaturated molecules called carotenoids are present in the natural diet and play an important role in different biological process, especially in reproduction. In order to gain more insights into these compounds in Nodipecten nodosus it was necessary to develop a suitable protocol for extraction of carotenoids from the gonads. Female gonads of cultured scallops (75 mm length) were lyophilized and macerated in liquid N 2 . To verify the effect of composition in organosolvents on the extracting solutions, two organic solvents were tested: acetone and hexane (Ac = O:Hex) at four ratios, 1:1, 1:3, 1:5, and 2:3, in four static extraction times: 0, 5, 10, and 15 minutes. Total carotenoids and astaxanthin contents were determined in the crude extracts by UV-visible spectrophotometry and high performance liquid chromatography (HPLC), respectively. Triplicate aliquots of 50 mg were used for each treatment. The results indicated that the best single extraction (0.312 ± 0.016 µg carotenoids/mg) was attained with Ac = O: Hex 1:3, for 15 minutes. Through exhaustive extraction methodology (10x), a superior yield (0.41 ± 0.001 µg carotenoids/mg) was obtained from a gonad sample in comparison to the highest value found for a single extraction. Astaxanthin content was reduced by 8.6% in carotenoid extract preservation assay, i.e., -18 °C, 26 days incubation, under N 2 atmosphere.Keywords: carotenoids, Nodipecten nodosus, extraction, organic solvents, scallop. Extração de carotenóides da gônada da vieira Nodipecten nodosus (Bivalvia: Pectinidae) ResumoEm moluscos bivalves marinhos, carotenóides insaturados estão presentes na dieta natural, com um importante papel em diversos processos biológicos, em especial na reprodução. A elucidação dos efeitos destes compostos em Nodipecten nodosus requer o desenvolvimento de um protocolo adequado para a extração de carotenóides das gônadas desses animais. Para isso, gônadas de vieiras cultivadas (75 mm de comprimento) foram liofilizadas e maceradas em N 2 líquido. Amostras em triplicata com 50 mg foram coletadas para a utilização em cada tratamento. Os conteúdos de carotenóides totais e astaxantina foram determinados via espectrofotometria de luz UV-visível e cromatografia líquida de alta eficiência (CLAE), respectivamente. O efeito da composição em organosolventes das soluções de extração foi testado utilizando-se acetona (Ac = O) e hexano (Hex) em quatro proporções (Ac = O:Hex): 1:1, 1:3, 1:5, e 2:3; em quatro tempos de extração: 0, 5, 10, e 15 minutos. Os resultados mostraram que o melhor rendimento de extração (0,312 ± 0,016 µg carotenóides/mg) foi obtido com Ac = O:Hex, 1:3, por 15 minutos. Com a utilização de protocolo de extração exaustiva (10x), uma quantidade superior (0,41 ± 0,001 µg de carotenóides/mg) foi obtida de amostras de gônada, comparativamente aos valores obtidos em extrações únicas. O conteúdo de astaxantina foi reduzido em 8,6% em testes de preservação deste metabólito em extratos crus (-18 °C, 26 dias de incubação em atmosfera de N ...

Effects of dietary astaxanthin and β carotene on gonadosomatic and hepatosomatic indices, gonad and liver composition in goldfish Carassius auratus (Linnaeus, 1758) broodstocks

“…After the sampling period, the frozen samples were dried at -55ºC (Christ Freeze Dryer-Alpha-1-2 LDplus model) and stored at -18°C in an amber flask filled with N2 for later carotenoid extraction (Sühnel et al, 2009).…”

“…For this analysis, 50 mg (dry tissue weight) samples were taken from each tissue type and added to 5 mL of an extracting solution containing pure acetone (Ac = O, analytical grade) and n-hexane (Hex, analytical grade) at a ratio (v/v) of 1:3, respectively. This mixture was then agitated in a magnetic agitator (10 min) at ambient temperatures (22°C) and under dark conditions (Sühnel et al, 2009). Carotenoid concentration in the extracted solution was determined according to procedures described by Sühnel et al (2009), a UV-visible spectrophotometer (Hitachi, U-1800) with the spectral window set between 380 and 750 nm was used, in triplicate.…”

Sexual stages of the female portion in the scallop Nodipecten nodosus (Linné, 1758) and astaxanthin quantity in each stage

“…Carotenoid extraction followed the methodology described in Sühnel et al (2009). The lyophylised sample was placed in a ceramic container with liquid nitrogen, mixed and 50 mg were removed.…”

“…The methodology used for the analysis is described in Sühnel et al (2009), where 300 µL of the carotenoid extracts were filtered (0.22 µ filter), and 10 µL of each sample was injected in a liquid chromatograph (Shimzadu LC -10A), equipped with a C 18 column of reverse phase Vydac (BioRad, 25 cm × 4.6 mm Reproductive cycle and astaxanthin levels in Nodipecten nodosus ∅) and UV-visible detector, operating with a reading of 470 ηm. A solution of acetonitrile and methanol (at a 90:10 proportion, TediaBrazil, HPLC degree) with a 0.8 mL/min flow was used as a mobile phase.…”

“…The gametes leave the paleal cavity through the urogenital pore (Román et al, 2001). Sühnel et al (2009) observed that the N. nodosus pectinid contains an unusually large amount of carotenoids in the female portion of the gonad. The present study quantifies the accumulation of astaxanthin in the female portion of the gonad of N. nodosus broodstock in each sexual stage, and provides a histological description of these different stages based on observation of individuals produced in the hatchery over the period of one year.…”