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Chemical Composition and Stimulating Effect of Citrus Hystrix Oil on Humans

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Chemical Composition and Stimulating Effect of Citrus Hystrix Oil on Humans FLAVOUR AND FRAGRANCE JOURNAL COMPOSITION AND EFFECTS OF CITRUS HYSTRIX OIL ON HUMANS 443 Flavour Fragr. J. 2007; 22: 443–449 Published online 18 July 2007 in Wiley InterScience (www.interscience.wiley.com) DOI: 10.1002/ffj.1820 Chemical composition and stimulating effect of Citrus hystrix oil on humans Tapanee Hongratanaworakit1* and Gerhard Buchbauer2 1 Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Srinakharinwirot University, Nakhon-nayok 26120, Thailand 2 Department of Clinical Pharmacy and Diagnostics, Centre of Pharmacy, University of Vienna, Althanstrasse 14, A-1090 Vienna, Austria Received 12 January 2007; Revised 15 May 2007; Accepted 28 May 2007 ABSTRACT: Kaffir lime essential oil was obtained from fresh peels of Citrus hystrix (Rutaceae) by hydrodistillation and analysed by GC and GC–MS. The effects of kaffir lime oil on human autonomic and behavioural parameters after massage were investigated in this study. Forty healthy volunteers participated in the experiments. Autonomic parameters recorded were skin temperature, pulse rate, breathing rate and blood pressure. Behavioural parameters were assessed by means of visual analogue scales (VAS). The kaffir lime oil caused a significant increase in blood pressure and a significant decrease in skin temperature. Regarding the behavioural parameters, subjects in the kaffir lime oil group rated themselves more alert, attentive, cheerful and vigorous than subjects in the control group. These findings are likely to represent stimu- lating/activating effects of the kaffir lime oil and provide some evidence for the use of kaffir lime oil in aromatherapy, such as causing relief from depression and stress in humans. Copyright © 2007 John Wiley & Sons, Ltd. KEY WORDS: physiological parameters; activating effect; behavioural evaluation; kaffir lime oil Introduction patients by increasing blood flow to the prefrontal cortex.10 Stress-relieving properties could also be attrib- The essential oil of kaffir lime (Citrus hystrix DC.; uted to C. sinensis oil, measuring the saliva cortisol con- family Rutaceae) is increasingly used as a fragrance in centration.11 An investigation at the Middlesex Hospital the food, perfumery and cosmetic industries. Kaffir lime in London showed that foot massage with the essential oil has been associated with anti-proliferative activity on oil from the flowers of C. aurantium (Neroli oil) KB and P388 cell lines and has the potential for cancer positively affects the psychological state of patients.12 treatment.1 The repellency effect of kaffir lime oil has Clinical aromatherapy for agitated behaviour in dementia been demonstrated by many researchers.2,3 Kaffir lime oil patients has been reported.13–16 Furthermore, aromatherapy shows antioxidant as well as free radical scavenging massage is increasing in oncology and palliative care activity.4–6 Antiviral activity of kaffir lime extract has unit.17–18 been reported by Khan et al.7 In medicine, interest in Recently there have been a variety of measurements the usage of citrus oils as therapeutically active agents to evaluate the physiological and psychological effects has grown considerably. Especially in aromatherapy, of fragrances, measuring such parameters as: changes Citrus sinensis (sweet orange oil) has been used as an in autonomic parameters, e.g. heart rate, breathing rate, antidepressant in the case of depression. Lehrner et al.8 blood pressure, eye-blinks, skin temperature and skin reported that ambient odours of C. sinensis oil reduced conductance19–23; changes in brain wave activities, e.g. anxiety and improved mood in patients waiting for electroencephalogram; contingent negative variation24–25; dental treatment. A mixture of citrus oils helps to reduce changes in mood, cognitive performances and emotion.26–28 the therapeutic doses of antidepressants in depressive A study conducted at the Human Cognitive Neuroscience patients.9 Aromatherapy massage using C. sinensis oil Unit, University of Northumbria, Newcastle upon Tyne, every week for 4 weeks improves depressive states in UK,29 showed that aromas of rosemary and lavender oils differentially affected cognition and mood in healthy adults. Analysis of performance revealed that lavender oil produced a significant decrement in the performance * Correspondence to: T. Hongratanaworakit, Faculty of Pharmacy, of working memory and impaired reaction times for Srinakharinwirot University, Rangsit-Ongkharak Road, Nakhon-Nayok both memory- and attention-based tasks compared to 26120, Thailand. controls. In contrast, rosemary oil produced a significant E-mail: [email protected] Contract/grant sponsor: Srinakharinwirot University, Thailand; Contract/grant enhancement of performance for overall quality of number: 012/2005. memory and secondary memory factors, but also Copyright © 2007 John Wiley & Sons, Ltd. Flavour Fragr. J. 2007; 22: 443–449 DOI: 10.1002/ffj 444 T. HONGRATANAWORAKIT AND G. BUCHBAUER produced an impairment of speed of memory compared β-pinene, γ-terpinene). The correction factors of the monoterpene to controls. This study indicated that these essential oils alcohols for the other components, i.e. linalool, exo-fenchol, can produce objective effects on cognitive performance neo-isopulegol, isopulegol and borneol, were obtained from the as well as subjective effects on mood. However, up to average of the correction factors of two standards monoterpene α now, no experiments on the effects of kaffir lime oil on alcohols (terpinene-4-ol, -terpineol). The percentage com- positions of the remaining components were calculated without the human autonomic nervous system and behavioural correction factors. measures after massage have been carried out. Therefore, the main objectives of the present study were: (a) to investigate the chemical composition of kaffir lime oil; Gas Chromatography–Mass Spectrometry (GC–MS) (b) to evaluate the effects of this fragrance compound on autonomic nervous system parameters and behavioural GC–MS was carried out on a Trace GC Ultra/Trace DSG parameters in healthy humans following transdermal Finnigan gas chromatograph fitted with a fused-silica BPX5 absorption. capillary column (30 m × 0.25 mm i.d., 0.25 µm film thickness); carrier gas, helium at 1.0 ml/min; injector temperature, 220 °C; split ratio, 1:20; transfer line temperature, 240 °C; column Experimental temperature, 60–246 °C at 3 °C/min; mass spectra obtained using an ionization energy of 70 eV and ion source temperature Fragrance Compound of 200 °C. The individual components were identified: (a) by comparison of their mass spectra with those stored in the MS database (NIST, WILEY libraries) and with mass spectra liter- Kaffir lime oil was obtained from the fresh fruit peel of 30 Citrus hystrix DC. by hydrodistillation for 2 h using a ature data; (b) by comparison of their linear retention indices Clevenger apparatus. The obtained oil was dried over anhydrous (LRI) on DB-5 column, calculated from the retention temper- ature/retention times of a homologous series of n-alkanes,31 with sodium sulphate and kept at 4 °C until analysed. The 32–35 average yield was 1.5% w/w. A voucher specimen has been those of authentic standards or with those of the literature. deposited at Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Srinakharinworit University, under Accession No. 4807295-/1307. Subjects Forty healthy volunteers aged 18–48 (mean 23.55 ± 6.06) years Gas Chromatography (GC) took part in the experiments. Demographic data for the control group and the experimental group are presented in Table 1. GC–FID was performed using a Perkin-Elmer AutoSystem Subjects were tested in individual sessions and randomly gas chromatograph fitted with a fused-silca PB-WAX capillary assigned to either the control group or the kaffir lime oil group. column (60 m × 0.25 mm i.d., 0.32 µm film thickness); column Each group consisted of 20 subjects. They were fully briefed, temperature, 90 °C for 4 min, then 90–220 °C at 10 °C/min and gave written informed consent to all aspects of the study held isothermal at this temperature for 3 min; injector and (Srinakharinwirot University Ethics Committee) and were free detector temperatures, 190 °C and 220 °C, respectively; carrier to withdraw at any time. Forty-eight hours prior to testing gas, helium at 1.0 ml/min; samples (1.5 ul) injected using a split subjects had to abstain from food, beverages and toiletries con- ratio of 1:10. Quantitative analysis of the oil component taining the essential oil of Citrus species or its constituents (e.g. was calculated according to the correction factor, using stand- limonene), i.e. citrus fruits, apricots, peaches, fruit juices, basil, ards at least for the main components, i.e. limonene, β-pinene, coriander, thyme, cinnamon, geranium and bergamot, as well as γ-terpinene, terpinene-4-ol, α-terpineol, since the components of from tea, coffee and alcohol. the same or similar chemical class could give a similar correc- tion factor. Therefore, the correction factors of the monoterpene hydrocarbons for the other components, i.e. α-thujene, α-pinene, Fragrance Administration camphene, sabinene, myrcene, α-phellandrene, α-terpinene and terpinolene, were obtained from the average of the correction In the experimental group, 1 ml 20% (w/w) solution of kaffir factors of three standard monoterpene hydrocarbons (limonene, lime oil in sweet almond oil was applied to the skin of the Table 1. Demographic data for the control and experimental groups Parameter Control group Kaffir lime oil group Number of volunteers 20 20 Sex (M:F) 8:12 9:11 Height (cm; mean ± SD) Male 172.75 ± 6.01 171.66 ± 4.27 Female 159.91 ± 4.48 159.45 ± 4.63 Weight (kg; mean ± SD) Male 61.87 ± 7.29 62.55 ± 9.11 Female 53.16 ± 4.70 52.63 ± 6.05 Copyright © 2007 John Wiley & Sons, Ltd. Flavour Fragr. J. 2007; 22: 443–449 DOI: 10.1002/ffj COMPOSITION AND EFFECTS OF CITRUS HYSTRIX OIL ON HUMANS 445 lower abdomen of each subject, and subjects themselves ness, mood and alertness) were used to assess behavioural massaged the oil into the skin for 5 min.
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