INTRODUCTION:

Safflower, Carthamus tinctorius L. is a member of the family Compositae or Asteraceae. It is cultivated mainly for its seeds, which yield edible oil. Traditionally, the crop was grown for its flowers, used for coloring and flavoring foods and making dye. The plant is known to possess many medicinal values. The seed oil offers quality edible oil which is rich in poly unsaturated fats (PUFA) The suitability of a vegetable oil for a particular use such as nutritional¹, industrial or pharmaceutical applications depends upon its fatty acid composition. However, the fatty acid composition of vegetable oil is highly variable in different plant species. In safflower oil, oleic and linoleic acids are the predominant fatty acids and can form more than 80% of the total in many varieties. The high levels of monounsaturated and polyunsaturated fatty acids (PUFA’s) improve the quality of the oil for human consumption. Studies have shown that it reduces blood cholesterol and plays an important role in preventing atherosclerosis²

The demand for beneficial PUFA’s has drastically increased in recent years, there are increasing efforts to find plant sources of PUFA’s that are economical and sustainable. In this way the source plant itself has attracted significant interest due to its high adaptability for dry climatic conditions with scanty rainfall³. It is a drought tolerant crop which is capable of obtaining moisture from levels not available to majority of crops⁴

Seeds of the species contain 22.5% oil on a whole seed basis. Quality of oil i.e. its composition is a significant concern for consumers, particularly the contents of oleic and linoleic acids which are proven as healthy sources of oil for human body. Safflower oil is thought to be one of the highest quality vegetable oils. The .approximate composition being - saturated acids palmitic acid ( 9%) & stearic acid (3%) mono unsaturated acids oleic acid (14%) and a large amount of poly unsaturated acids ( linoleic acid (72%) .

There are two types of safflower varieties, the type that produces oil which is high in monounsaturated fatty acids (oleic acid), and those with high concentrations of polyunsaturated fatty acids (linoleic acid)⁵.The present study was carried out to identify parental lines with sufficient genetic diversity for these biochemical parameters and examine its exploitable variability in the fatty acid profile in twenty six different lines of Carthamus species.

MATERIALS AND METHODS:

Seeds of plants of the following safflower germplasms were used in the present study, obtained from local agriculture university. Two trials (IET and CVT) of the national standard variety A-1 was included for analysis of oil content and fatty acid composition. Other germplasms lines included for study were

TM-29, CSS 611 (SVT), CTV 73, CSS158(SVT), CTV 78, 6440 (SVT), CSS 111 (SM), 499 (SM), NDS-1,CSS 611, CSS 19-1 (SM), 6440 (SM), 8304 (SM), CTV 26, T-65, CTH-1, CTV 9, CTV 2, CTV 3, CTH-2, CSS 98 (SM), CSS 19 (SM), CTV 84, and CTV 37.

After sampling the seeds were subjected for chemical analysis.

Chemical analysis:

Removal of moisture: Seeds were dried in oven at 60 C for 10 hrs. to remove the moisture. Oil content estimation was done on moisture free seeds.

Oil content: The estimation of oil content was done on oxford 4000 NMR- Analyzer one single point calibration in duplicate. The operational parameters are given below:-

Instrument Oxford 400 NMR analyzer

Version NA-01-9T

Mode 4

R.F Level 350

A F Gain 500

Gate width 1.5

Analytical time 10

Repeats 1

Calibration time 10

Standard sample for oil content estimation was prepared by extracting oil by soxhlet method using 40 – 60 petroleum ether as solvent.(6,7)

Determination of Fatty Acid Profile

It involved two stages. The first is the sample preparation and second stage is separation of different fatty acids by subjecting the sample ester to Gas- liquid chromatography.

(a) Sample preparation:

Seeds were bulked and 5 g clean and mature seed samples taken and crushed in a porcelain pestle mortar and washed with Petroleum ether (40 -60 B.p) to extract oil in cold. Excess of solvent was removed by keeping the oil overnight. Fatty acid methyl esters (FAMEs) were prepared according to the standard AOCS methods⁶ .The fatty acid composition was reported as a relative percentage of the total peak as following:

Area under individual peak

Percentage of total fatty acid =----------------------------------

Total area of all the peaks

RESULTS AND DISCUSSIONS:

Oil content:

The oil content is estimated by NMR and the values as depicted in Table 1 below showed wide range of variations. The values ranged from 24.79 % in SM CSS 19-1 to 37.6 % in SM NDS-1.The national standard variety was shown to have an oil content of 29.9 % and 30.07 % in IET and CVT respectively, while local check had 30.4 %..The oil content of seeds depends on many factors like genetic factors, agro-ecological conditions. The magnitude and range of the values of seed oil were similar to those reported previously in several Indian cultivars⁸. In addition to genetic factors; some environmental conditions may also influence variations of this parameter⁹.

Table 1 Oil content of various lines of safflower

	Line	Oil content	Line	Oil content
	TM-29	34.0	SM 8304	27.9
SVT CSS 611		34.3	CTV 26	29.0
IET A-1		30,0	T-65	30.4
IET CTV 73		31.7	CTH-1	28.9
SVT CSS 158		33.4	CTV-9	26.0
CTV 78		35.5	A-1CVT	29.9
SVT 6440		29.2	CTV-2	28.9
CSS 111		28.6	CTV-3	30.1
SM 499		31.3	CTH-2	31,7
SMNDS-1		37.6	SM CSS-98	33.2
SM CSS 611		26.5	SM CSS 19	36.5
SM CSS 19-1		24.8	CTV 84	36.5
SM6440		32.2	CTV 37	34.7

Fatty acid profile:

The oils extracted from the accessions were analyzed by gas liquid chromatography to determine their fatty acid composition. Four major fatty acids¹⁰ found were palmitic (16:0), stearic (18:0), oleic (18:1) and linoleic acid (18:2) . Oleic and linoleic acids are the major fatty acids of all the oil samples¹¹. In certain cases myristic acid was also observed to be present as a minor component. In general linoleic acid was in the highest concentrations, whereas stearic acid was in minimum amounts. Table below shows the fatty acid composition of all the samples evaluated

The highest percentage of palmitic acid is shown by CTV 9 and the lowest by the variety SM CSS 98.The respective amounts being 12.5 % and 4.1 %.The national standard variety A-1 had 6.1% in IET and 12.4% in CVT. a variation of 6.3% was shown. The local standard had 11.0 % palmitic acid.

The percentage of stearic acid varies from 0.7 for TM-29 to 7.1 for A-1 (CVT).Local check T-65 had 3.1 % stearic acid.

From the percentage listed in Table 2, it is clear that the range of oleic acid is from 8.1% to 24.3 % for SVT 6440 and CTH 2.The national standard A-1 had 17.0% and 13.6% in IET and co-ordinate trial variety. Six varieties had more than 18% oleic acid. The local standard T-65 had 11.9 % oleic acid. Line CTH-2 having a high percentage of oleic acid can be further explored for developing high oleic varieties.

Table 2 Fatty acid composition (area% ) of seed oil of different lines of Safflower

Line	Myristic Acid	Palmitic acid	Stearic acid	Oleic acid	Linoleic acid	Saturated Acid	Unsaturated acid
Line	Myristic Acid	(16:0)	(18:0)	(18:1)	(18:2)	Saturated Acid	Unsaturated acid
TM-29	-	12.4	0.7	9.4	77.3	13.1	86.7
SVT CSS 611	-	11.4	4.5	10.9	72.7	15.9	83.6
IET A-1	0.68	6.1	3.0	17.0	73.2	9.78	90.2
IET CTV 73	1.4	12.1	2.3	10.1	75.3	15.8	85.4
SVT CSS158	0.5	6.8	2.1	11.3	78.3	9.4	89.6
CTV 78	1.7	11.7	7.0	14.6	64.9	20.4	79.5
CSS 111	-	7.1	1.9	18.2	73.1	9.0	91.3
SVT 6440	1.1	8.2	1.8	8.1	80.2	11.1	88.3
SM 499	0.2	4.2	3.2	12.5	79.8	7.2	92.5
SMNDS-1	1.5	8.6	2.9	20.7	65.9	13.0	86.6
SM CSS 611	1.3	12.2	5.1	15.9	65.2	17.3	81.1
SM CSS 19-1	1.5	7.6	1.2	17.2	68.9	10.3	86.1
SM6440	-	8.1	5.2	20.2	66.5	13.3	86.7
SM 8304	0.9	6.9	1.3	18.3	70.1	9.1	88.4
CTV 26	-	11.8	4.4	18.1	65.6	16.2	83.7
T-65	1.0	10.0	3.1	11.9	73.8	14.1	85.7
CTH-1	-	9.5	2.8	9.1	78.1	12.3	87.2
CTV-9	1.0	12.5	1.5	11.7	73.2	15.0	84.9
A-1CVT	1.6	12.4	7.1	13.6	65.2	21.1	78.9`
CTV-2	1.1	9.6	5.1	10.1	72.6	15.8	82.7
CTV-3	-	12.0	2.8	11.5	73.5	14.8	85.0
CTH-2		6.9	2.1	24.3	65.5	9.0	89.8
SM CSS-98	-	4.1	2.7	17.1	76.1	6.8	93.2
SM CSS 19	1.0	10.5	2.0	9.2	77.3	13.5	86.5
CTV 84	1.2	8.0	4.2	15.4	71.2	13.6	86.4
CTV 37	-	9.4	3.1	17.1	69.5	12.5	86.6

As expected linoleic acid is present in highest concentration among all the fatty acids ranging from 64.9 % to 80.2 % for CTV 78 and SVT 6440.The corresponding values for A-1, National check is 73.2 % and 65.2 % in IET and CTV. There was least variation in the linoleic acid concentration of the two trials of national check. The percentage of saturated fatty acids varies from 21.1 % for national check A-1 to 6.8% for SM CSS-98 .The unsaturated fats percentage being as high as 93.2 % for SM CSS -98.

The high level of unsaturated fatty acids increases the quality of the oil for human consumption. The accessions with the highest oil content were relatively richer in the linoleic acid content.

Relationships among Fatty Acid Compositions

The results of correlation analyses among fatty acids of the accessions examined are as follows: Palmitic acid was significantly positively correlated with stearic and oleic acid. This result agrees with the previous report on sesame¹². Stearic acid was significantly positively correlated with oleic acid while it was significantly negatively correlated with linoleic acid. This association is well documented and reported in sesame¹³.

CONCLUSION:

This study provides an important insight for improving and developing new varieties with both high oleic and linoleic acid content. The studies on fatty acid compositions of several germplasms collections of crop plants have exposed wide variations in the proportions of saturated and unsaturated fatty acids, offering possibilities of developing superior quality edible oils and specialized industrial oils

One reason for the inverse relationship between these fatty acids could have been dependent on the environment where the genotypes were grown however, correlations between oleic and linoleic acids in many crop plants were always strongly negative. There should be a genetic contribution for the related fatty acids.

Knowles¹⁴ determined the genes involved in the inverse relationship between oleic and linoleic acid in safflower as pointed out by Johnson et al.¹⁵.

The results obtained in this study provide useful background information for developing new cultivars with high oil content and modified fatty acid compositions.

REFERENCES:

1. Baker M.L.et al. Nutritional value of undecorticated safflower meal. Nebraska Agriculture Experiment .Station. Bulletin No.402; 1951.

2. Ghafoorunissa. Dietary fats/oils and heart diseases. Sustainability in oil seeds. Edited by Prasad, M V R Indian Society of Oil Seeds Research, Hyderabad, 1994 : 436–49

3. Bassil, E.S., Kaffka, S.R., Response of safflower (Carthamus tinctorius L.) to saline soils and irrigation: Consumptive water use. Agriculture Water Management, 54 ; 2002: 67- 80.

4. Gawand, P.B.et al.,. Evaluation of productivity of safflower cultivars under moisture and nutrient management in rainfed vertisols. VIth International Safflower Conference Proceedings, 2005: 205-209.

5. Horowitz,B.,.Winter,G. High oleic type safflower varieties. Nature 170; 1937: 582-583

6. AOCS,.Official methods and recommended practices (1993). The American Oil Chemists Society, Champaign, IL

7. Madsen,E. Nuclear Magnetic Resonance Spectrometry as a quick method for determination of oil content in rapeseed Journal of American Oil Chemists Society 53(7); 1987: 467-469.

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10. Penumetcha, M. et al. Dietary oxidized fatty acids: an atherogenic risk. Journal of Lipid Research, 41; 2000: 1473-1480

11. Lee,Y C et al., Chemical composition and oxidative stability of safflower oil prepared from safflower seed roasted with different temperatures. Food Chemistry 84 ; 2004 : 1–6.

(12). Brar G.S, Variations and correlations in oil content and fatty acid composition of sesame. Indian Journal of Agricultural Science 52; 1982 :434–439.

12. Mandal S, et.al (2002) Correlation studies on oil content and fatty acid profile of some Cruciferous species. Genetic Resources and Crop Evolution 49; 2002 :551–556.

13. Knowles PF, Modification of quantity and quality of safflower oil through plant breeding. Journal of American Oil Chemists Society 46; 1969 :130–132.

14. Johnson R C, et al, Oil and meal characteristics of core and non-core safflower accessions from the USDA collection. Genetic Resources and Crop Evolution 46; 1999 :611–618.

Received on 17.09.2012 Modified on 03.10.2012

Asian J. Research Chem. 5(10): October, 2012; Page 1289-1292