The fatty acid profiles – including petroselinic and cis-vaccenic acid – of different Umbelliferae seed oils

1 Introduction

In the development of new oil seed crops interest has turned to the members of the Umbelliferae family. The yieldsper hectare (4–30 dt/ha) and oil content of these oil crops(8–24%) are not extremely high, and the properties of culti-vation are not as good as of other well-known seeds like sun-flower or rapeseed. But these agricultural crops, which con-tain spice plants like caraway, celery, coriander, dill, andparsley, are of interest due to the high amount of petroselinicacid [1, 2].

On the one hand this octadecenoic acid (C18:1 c6) is usedas raw material for fine chemicals, on the other hand itscleavage products – adipic and lauric acid – obtained by oxidative ozonolysis are used for technical purposes:

Adipic acid is used for the production of softeners andnylon. Lauric acid (C12:0), utilized as raw material for soft-

eners, emulsifiers, detergents, and soaps, is the major fattyacid of palm, coconut, and babassu oil, so Europe has toimport this raw material for the oleochemical industry fromSoutheast Asia [3, 4].

However, at the present time the inexpensive supply fromboth lauric and adipic acid does not favour the commercialdevelopment of oilseed crops bearing petroselinic acid [5].

We have selected three Umbelliferae seeds – fennel, cara-way, and coriander – in different varieties in order to analyzethe fatty acid profiles of the oils. The only comprehensivestudy of fatty acid profiles of Umbelliflorae seed oils (con-tains the family Umbelliferae) was performed by Kleimanetal. [6] in 1982. But the method used for the determination ofthe fatty acid profiles lends itself only to basic investiga-tions, not to routine analysis.

In general, fatty acid patterns of vegetable oils are deter-mined and quantified as fatty acid methyl esters by capillary-gas chromatography [7]. Two further positional isomers ofoctadecenoic acid, oleic acid (C18:1 c9), and cis-vaccenicacid (C18:1 c11) occur in Umbelliferae seed oils. The sepa-

498 Fett/Lipid 100 (1998), Nr. 11, S. 498–502 © WILEY-VCH Verlag GmbH, D-69451 Weinheim, 1998 0931-5985/98/1111-0498$17.50+.50/0

Forschungsbeiträge/Research Papers

Different varieties of fennel, caraway, and coriander (3 fennel, 7 cara-way, and 4 coriander varieties), which belong to the Umbelliferae fami-ly, were analyzed on oil and water content, subsequently the fatty acidprofiles of the oils were determined by automated gas chromatography.Using fatty acid butyl esters a complete fatty acid profile including petroselinic and cis-vaccenic acid was obtained.Furthermore, the obtained fatty acid contents were compared to thosecited in literature, which were determined using different analysis pro-cedures.

The fatty acid profiles – including petroselinic and cis-vaccenic acid – of different Umbelliferae seed oils

Birgit Reiter*, Marion Lechner*, and Eberhard Lorbeer*

Das Fettsäuremuster einschließlich Petroselin- und cis -Vaccen-säure aus verschiedenen Samen der Umbelliferae. Aus der Fami-lie der Doldengewächse wurden unterschiedliche Sorten von Fenchel,Kümmel und Koriander (3 Fenchelsorten, 7 Kümmel- und 4 Koriander-sorten) auf ihren Öl- und Wassergehalt untersucht. Das Fettsäuremusterder Öle wurde mittels automatisierter Kapillar-Gaschromatographie bestimmt. Durch die Derivatisierung der Triglyceride zu Fettsäurebutyl-estern konnte das vollständige Fettsäurespektrum einschließlich der Petroselin- und der cis-Vaccensäure ermittelt werden.Die Daten wurden mit Literaturwerten, die durch unterschiedliche Be-stimmungsmethoden erhalten wurden, verglichen.

* Institute of Organic Chemistry, Vienna, Austria.

Tab. 1a. Comparison of petroselinic, oleic and cis-vaccenic acid content (in %) of fennel (Foeniculum vulgare) seed oil by different analyticalprocedures.

analysis procedure reference C18:1 c6 C18:1 c9a C18:1 c11a

ozonolysis/GLC Kleiman et al. [6] 72.9 8.7 –

TLC/prep.GLC/ozonol. Seher et al. [11] 70.4 2.6 0.5

TLC/densitometry Nikolova et al. [12] 70.9 8.6 –

KMnO4-cleavage/GLC Mbayhoudel[13] 78.0 8.2 –

GLC/13C NMR Mallet et al. [14] 73.5 – –

Charvet et al. [15] 77.9 7.7 –13C NMR Gunstone [16] 75.3 5.9 –

GLC:v TMS-derivatives Ucciani et al. [17] 76.6 5.5 –

v isopropylester Wolff et al. [18] 78.9 5.1 0.2

a if no value, not analyzed.

ration of these isomers as methyl esters is normally insuffi-cient for quantitative analysis [8]. Griffiths et al. [9] claimedthat the gas chromatographic separation of the fatty acidsmentioned above as methyl ester with a capillary columncoated with a highly polar stationary phase is sufficient forquantitative analysis, but under similar conditions theseresults could not be repeated in our laboratory [10].

Tabs. 1a, 1b, and 1c show a comparison of the methodsused for the determination of petroselinic acid and, if ana-lyzed, of oleic and cis-vaccenic acid. All showed derivativesand procedures are either laborious and time-consuming orseparation efficiency is not sufficient for proper quantification.

However, in most cases complete fatty acid profiles aremissing, especially values of the cis-vaccenic content, andno investigations on different varieties were carried out.

The aim of this work is to determine the fatty acid patternof selected seed oils by gas chromatography with a methodrecently published by Reiter et al. [21], to compare theresults with literature data and to discuss the variation ofpetroselinic acid content within a species.

2 Materials and Methods

2.1 Reagents and samples

Methyl esters of petroselinic, oleic, cis-vaccenic, palmit-ic, linoleic and margaric acids (for use as standards) werepurchased from Sigma (Deisenhofen, Germany), Sodium-tert-butylate from Fluka (Buchs, Switzerland). The solventsn-hexane and n-butanol were used in p.a. quality. Caraway,fennel and coriander seeds were purchased from Austrianand German suppliers (see Tab. 2 footnotes). Seeds werecrushed, and the oil was extracted with petroleum ether in aSoxhlet extractor. The water content in the seeds, and the oilcontent in the dry matter were determined using the DGFstandard methods B-I 4 (88) and B-I 5 (88) [22] (Tab. 2).

2.2 Preparation of fatty acid butyl ester

Preparations of sample solutions were carried out accord-ing to Reiter et al. [21]: Sodium-tert-butylate in butanol(500µl, 0.1 M) was added to a mixture of 2 mg vegetable oiland 295µl of the solution of the internal standard margaric

acid methyl ester (6.11 mg margaric acid methyl ester/ml n-hexane). After 10 min at room temperature 500µl of a 5%aqueous solution of NaHSO4 were added. Part of the butanolphase (300µl) was transferred into a vial, the solvent wasremoved in a stream of nitrogen and the butyl esters weretaken up in 3 ml n-hexane.

2.3 Instrumentation

Gas chromatographic analyses were carried out on a GC8000 gas chromatograph (Fisons Instruments, Milano,Italy). The instrument was equipped with a flame-ionisationdetector, an on-column injector, an autosampler (AS 800,Fisons Instruments, Milano, Italy) and an uncoated fused-

Fett/Lipid 100 (1998), Nr. 11, S. 498–502 499

Tab. 1b. Comparison of petroselinic, oleic and cis-vaccenic acid content (in %) of caraway seed oil (Carum carvi) by different analytical proce-dures.


ozonolysis/GLC Kleiman et al. [6] 35.4 24.1 –

TLC/prep.GLC/ozonol. Seheret al. [11] 42.0 14.0 0.8

13C NMR Gunstone[16] 40.3 17.0 –


Tab. 1c. Comparison of petroselinic, oleic and cis-vaccenic acid content (in %) of coriander (Coriandrum sativum) seed oil by different analyticalprocedures.


TLC/prep. GLC/ozonol. Seher et al. [11] 36.0 44.0 0.8

TLC/densitometry Nikolova et al. [12] 64.9 5.0 –

GLC:

v methyl ester Griffiths et al. [9] 71.8 – –

v methylthio derivatives Thies [19] 91.0 1.0 0.6

v butyl ester Thies [20] 74.7 5.8 1.0


Tab. 2. Table of the analyzed seed oils and their origin and source ofsupply.

no. name variety water content total oil[% in the [% in the seed] dry matter]

1 Fennel1 Berfena 5.4 14.12 Fennel1 – 6.9 10.53 Fennel2 Magnafena 6.2 14.6

4 Caraway1 Bleya 7.3 14.35 Caraway2 Israel 7.1 12.36 Caraway2 – 6.0 9.57 Caraway1 Plewiski 6.4 13.28 Caraway1 Szilas 6.9 12.19 Caraway2 Volhouden 6.4 12.410 Caraway1 Volhouden 6.3 10.8

11 Coriander3 Corry 7.0 14.012 Coriander4 – 6.5 12.513 Coriander1 – 7.1 16.014 Coriander5 – 7.2 9.2

1 SBL – Saatbau Linz, Linz, Austria.2 LCH-BA – Bundesanstalt und Forschungszentrum für Landwirt-

schaft, Vienna, Austria.3 BORRIES – W. Borries-Eckendorf Pflanzenzuchtbetrieb, Leopolds-

höhe, Germany.4 AUSTROSAAT, Vienna, Austria.5 FAL – Bundesforschungsanstalt für Landwirtschaft, Braunschweig-

Völkenrode, Germany.

silica precolumn, deactivated with DPTMDS (diphenyltetra-methyldisilazane), (2 m × 0.53 mm i.d., BGB Analytik,Rothenfluh, Switzerland), which was connected in series to a30 m × 0.32 mm ID fused silica column coated with a0.25µm film of DB-23 (J&W Scientific, Folsom, CA, USA).By means of the autosampler 1µl of the sample was injectedat 70 °C. Then oven temperature was heated at 20 °C/min to110 °C, then at 3 °C/min to 200 °C. Hydrogen was used ascarrier gas at a flow rate of 2.6 ml/min. The detector tem-perature was held at 240 °C.

A personal computer and the software program Chrom-card for Windows (Fisons Instruments,Milano, Italy) wasused for data acquisition and evaluation.

2.4 Calibration

Standards of fatty acid butyl esters (BE) are not commer-cially available, so fatty acid methyl esters (ME) wereobtained and transesterified to butyl esters. Triglycerides ofthe fatty acids in the oil were similarily transesterified tofatty acid butyl esters.

A standard solution containing known amounts of thereference substances in n-hexane (petroselinic acid ME1.14 mg/ml, oleic acid ME 0.24 mg/ml, palmitic acid ME0.18 mg/ml, and linoleic acid ME 0.65 mg/ml) and an inter-nal standard solution with margaric acid ME in n-hexane(6.11 mg/ml) were transformed to their butyl esters and analyzed three times each by automated GC in order todetermine the ratio Ac/Ast (peak area of component (Ac)/peak area of internal standard (Ast)).

Good linearity of the calibration functions was observedwithin the concentration range of interest. Tab. 3 shows cali-bration functions for petroselinic acid, oleic acid, linoleicacid and for cis-vaccenic, stearic, and palmitic acid. Cis-vaccenic, stearic, and palmitic acid have about the same con-centration in the seed oil, so the same response factor was used.

2.5 Quantitative analysis

The concentration of palmitic, stearic, petroselinic, oleic,cis-vaccenic, and linoleic acid were determined in caraway,coriander, and fennel seed oil using margaric acid ME asinternal standard. Concentrations were calculated based onthe following equations:

Cc = Cst × [Rf × (Ac/Ast) + coeff.]; (Cc – concentration offatty acid as butyl ester in the sample (mg/ml); Cst – concen-tration of the internal standard (mg/ml); Rf – response factorof fatty acid).Cfa = (Cc/Ct) × 100; (Cfa – concentration of fatty acid (%); Ct – total of all Cc (mg/ml)).

3 Results and Discussion

3.1 Oil and water content

Due to the low water content of the seeds variations inextraction yields cannot be attributed to the water content,but rather depend on seed maturity. Oil contents of all inves-tigated specimens are constant, but differ considerably within the species: fennel 10.5–14.6%, caraway 9.5–14.3%,coriander 9.2–16% (see Tab. 2).

3.2 Sample treatment and gas chromatographicanalysis

Transesterification of triacylglycerols to fatty acid butylesters is a simple one-step reaction similar to the formationof fatty acid methyl esters, and lends itself to the simul-taneous preparation of a number of samples.

The best separation between petroselinic and oleic acidbutyl esters was observed using the gas chromatographicconditions described above. Fig. 1 shows chromatograms ofcaraway, coriander, and fennel seed oils, analyzed as fattyacid butyl esters with margaric acid as internal standard. Be-tween petroselinic and oleic acid butyl ester nearly baselineseparation was achieved. cis-Vaccenic acid was completelyseparated from petroselinic acid.

Tabs. 4, 5 and 6 show the fatty acid profiles of fennel,caraway, and coriander seed oils.

3.3 Comparison of data obtained

Fennel seed oil:In fennel seed oils the values of petroselinic acid content

obtained (71.9–73.9%) (see Tab. 4) are within the range citedin literature (70.4–78.9%). The contents of all other fattyacids have standard deviations lower than 0.5%.

Caraway seed oil:Caraway seed oil (Tab. 5) shows some differences: Except

for two varieties (33.5 and 36.4% petroselinic acid content,respectively) the range of content in petroselinic acid contentis quite narrow (41–42.7%). Data found in literature show awider range in petroselinic acid content (35.4–40.3%).

Coriander seed oil:Tab. 6 shows the fatty acid profile of the coriander seed

oils. Except for two values for petroselinic acid cited in literature (a high value of 91% [19] and a rather low value of36.0% [11]) the range of variation in petroselinic acid con-tent obtained by our analysis (64.9–74.7%) is quite similar

500 Fett/Lipid 100 (1998), Nr. 11, S. 498–502

Tab. 3. Calibration functions for petroselinic acid, oleic acid and cis-vaccenic acid.

Rf coeff. SE r2

petroselinic acid 1.07 – 0.012 0.996oleic acid 1.21 –0.0067 0.003 0.994cis-vaccenic, palmitic, stearic acid 1.02 – 0.002 0.997linoleic acid 1.14 – 0.006 0.996

Abbreviations: Rf – response factor; coeff. – coefficient; SE – standard error; r2 – mean-square error.

Tab. 4. Fatty acid profile of three different varieties of fennel (Foeniculum vulgare) seeds.

fatty acid composition [% total oil]no. name variety C16:0 C18:0 C18:1 c6 C18:1 c9 C18:1 c11 C18:2

1 Fennel Berfena 4.3 0.9 73.7 4.8 0.3 16.02 Fennel – 4.3 1.0 73.9 4.6 0.3 15.83 Fennel Magnafena 4.4 1.4 71.9 5.3 0.4 16.5

mean 4.4 1.2 72.9 5.0 0.4 16.2SD 0.1 0.2 1.0 0.4 0.1 0.4

to that found in literature (67.1–73.0%). The low value,which is accompanied by a high oleic acid value partlyagrees with the investigations of Lashminarayana et al. [23]:The increase in petroselinic content is followed by adecrease in palmitic and oleic acid content during matura-

tion. It seems possible that this oil was extracted from im-mature seeds. The extremely high value of petroselinic acidobtained by Thies [19] may be explained by an insufficientseparation of methylthio derivatives of fatty acids, whichdoes not allow an exact quantification. In general, oil plants

Fett/Lipid 100 (1998), Nr. 11, S. 498–502 501

Tab. 5. Oil content and fatty acid profile of seven different varieties of caraway (Carum carvi) seeds.


4 Caraway Bleya 4.2 1.1 42.5 15.4 0.7 36.25 Caraway Israel 4.7 1.5 36.4 20.8 0.7 35.96 Caraway – 4.7 1.6 41.0 17.0 0.7 35.07 Caraway Plewiski 4.2 1.2 42.0 15.4 0.6 36.58 Caraway Szilas 4.2 1.2 42.4 15.2 0.6 36.39 Caraway Volhouden 4.0 1.3 42.7 16.6 0.6 34.810 Caraway Volhouden 4.7 1.7 33.5 24.0 0.7 35.4

mean 4.5 1.4 40.5 17.1 0.7 35.9SD 0.3 0.2 2.4 2.2 0.0 0.6

Tab. 6. Oil content and fatty acid profile of four different varieties of coriander (Coriandrum sativum) seeds.


11 Coriander Corry 4.4 1.1 67.2 8.1 0.7 18.512 Coriander – 4.0 0.6 67.8 8.5 0.7 18.413 Coriander – 3.2 0.7 73.0 8.2 0.7 14.214 Coriander – 4.1 0.9 67.1 9.2 0.8 17.9

mean 3.8 0.7 69.3 8.6 0.7 16.8SD 0.4 0.1 2.6 0.4 0.0 1.9

Fig. 1. Chromatograms of (A) caraway, (B) coriander and (C) fennel seed oil, analyzed as fatty acid butyl esters (BE) with margaric acid as internal standard peak: 1 = palmitic acid BE, 2 = stearic acid BE, 3 = petroselinic acid BE, 4 = oleic acid BE, 5 = cis-vaccenic acid BE, 6 = linoleic acid BE, I.S. = internal standard margaric acid BE.

(A) (B) (C)

– exept of genetically modified or breeded – are not able toaccumulate more than 80 % of one fatty acid due to oilcropbiochemistry [5].

3.4 Conclusion

The data obtained by the method reported above agreewith results described in literature. Minimized sample treat-ment and the possibility of complete automatization allow ahigh sample throughput within a short time.

The determination of the complete fatty acid profile usingbutyl esters is performed as fast as the standard method forfatty acid analysis [7], and due to the improved resolutionadditional information, especially the contents of petroseli-nic and cis-vaccenic acid, is obtained from single gas chro-matographic runs.

Acknowledgements

The authors thank G. Benesch, N. Wimmer, and M. Zillinger for prac-tical support. This work was financially supported by the AustrianFederal Ministry of Agriculture and Forestry (Project Nr. L 896/94).

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Address of the authors: Mag. Birgit Reiter,Mag. Marion Lechner,Dr. Eberhard Lorbeer, University Vienna, Institute of Organic Chem-istry, Währingerstr. 38, 1090 Vienna, Austria.

[Received: March 5, 1998; accepted: August 12, 1998].

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Documents

The fatty acid profiles – including petroselinic and cis-vaccenic acid – of different Umbelliferae seed oils