Abstract
Hydrosol soft drinks in Persian nutrition culture are produced as side products of the essential oil industry to be used as safe remedies for treatment of some ailments. This study investigated hydrosols for women’s hormonal health conditions. Detailed information was gathered by questionnaires. Chemical constituents of these mono- or poly-herbal hydrosols were identified after liquid/liquid extraction and gas chromatography–mass spectrometry. Hierarchical cluster and K-means analysis (SPSS software) were used to find their relevance. A literature survey was also performed. In most cases, thymol, carvacrol, and carvone were the major constituents except for dill, white horehound, willow, Moderr, and yarrow hydrosols, whose their major components were dill ether, menthol, phenethyl alcohol, linalool, or camphor. Based on clustering methods, some similarities could be found in their constituents with some exceptions. None of them have been studied scientifically before. These investigations may lead to the development of some functional drinks or even new lead components.
Different ethnomedicinal herbal formulations have long been used by women to treat hormonal and reproductive health conditions such as premenstrual syndrome, menopausal symptoms, hormonal imbalance, infertility, or as contraceptives. 1,2 In many communities, because of economic or cultural issues, herbal remedies are the most—or even the only—available therapeutics. Despite the necessity of evaluating their purity, safety, efficacy, and authenticity, herbal formulations are not currently subjected to the same regulations as conventional drugs, which is due to a lack of knowledge about their constituents. 3 Another problem arises from misadministration and lack of knowledge about the side effects. For example, many herbal formulations—which may be used for other therapeutic properties or even ingested as daily food or drinks—may cause unwanted side effects for a pregnant woman or her fetus. 4 Scientific investigation of safety and efficacy of herbal remedies, food, and beverages with potential effects on hormonal condition may prevent a notable number of unwanted infertility cases, abortions, or fetal abnormalities. It also can lead researchers to reach new active components as well as functional food or beverages for use as contraceptives or fertility therapeutics or supplements. 5,6
In Iranian nutrition culture as well as Persian ethnomedicine aromatic waters, or Araghijat or Araghiat (plural of Aragh in Persian), are consumed as delicious daily drinks or as functional beverages. They are usually sweeten with natural sugars such as sucrose or honey. Some prepared syrup of aromatic waters can also be found in food markets containing some additives such as colors, flavoring agent, and sweeteners to improve the organoleptic properties.
Aromatic waters, which are also called floral water, distillate water, or hydrosols, are the side products of the volatile oil industry. 7 During industrial hydrodistillation water is evaporated simultaneously with the essential oil of the plants. After condensation of the vapors in contact with cold vessels or tubes, the liquefied components are separated into 2 phases inside a collecting vessel, an oily phase and the aromatic water saturated with different amounts of the volatile components of the plant, which are partly or completely soluble in water. 8,9 These 2 phases are then collected; the oily phase (essential oil) is usually sold to the pharmaceutical or cosmetic industry while the aromatic water, depending on its taste, potency, and biological properties, is diluted 1:8 or 1:12 with water. For marketing purposes, the aromatic waters might be distributed in big containers (250-1000 liters) for retail shops or in small sealed polyethylene terephthalate or glass containers (1-5 liters). Some are pasteurized before marketing. In traditional and folk Persian medicine, aromatic water drinks are used also for medicinal purposes to treat different conditions. Despite some adverse effects in improper applications, they are considered as safe beverages. They are mostly mono-herbal but some poly-herbal hydrosol (Aragh) can be found in the food market. 10,11 Depending on the plants used to prepare each of the aromatic waters, an overall nature is considered including, hot, cold, wet, dry, or moderate. Pure essential oils are very potent or even harsh in presenting their medicinal activities and are not usually safe in oral administration. But these hydrosols have their unique aroma and composition, which is considerably different from the pure essential oil they co-distilled with. They are usually moderated and balanced by the water and their water-soluble volatile components. 12,13 On the other hand, aromatic waters have additional properties not possessed by the essential oils alone.
Over 50 different types of hydrosol beverages are manufactured and marketed in Persian nutrition culture, but as far as we know, chemical composition and biological activities of many of them have not been investigated scientifically. This study was designed to investigate the chemical composition of aromatic waters and hydrosol beverages used in Persian folk medicine for women’s hormonal and reproductive health conditions. A wide range of plants are used in Persian folk medicine for these conditions, but only those plants were investigated that are used to prepare hydrosol beverages.
Materials and Methods
An Overview of Geographical Profile and Climate Variation of Field Study
Fars Province with the highest production rate for aromatic waters was selected as the field of study. Fars, or known in Old Persian as Pârsâ, is the original homeland of the ancient Persians.
The province, with area of 122 400 km2 and a population of 4.59 million people, is located in the south of Iran. There are 3 different climates in the Fars province: the highland area with moderate cold winters and mild summers; the central regions, with relatively rainy mild winter and hot dry summers; and the south to southeast region, which has cold winters with hot summers. The average temperature of Shiraz as the province administrative center is 16.8°C (ranging from 4.7°C to29.2°C). 14
The geographical and climatic variation enriches the province with varieties of plants that has a huge influence on agricultural and herbal industries. Over 84 manufactories in Fars province produce different aroma water beverages with full industrial techniques (19 manufactories) or traditional (65 manufactories) equipment. These manufactories are mostly located in Meymand and Darab cities, and their products are distributed in retail markets all over the country.
Information and Sample Collection
To gather information about different aromatic beverages used in Persian folk medicine for women’s hormonal and reproductive health conditions, a field study was conducted from June 2013 to June 2014 under the supervision of a local researcher as a native guide in all visits. A suitable questionnaire was filled according to the information gathered in all visits to local manufactories or their shops. However, most popular aromatic waters with indications for women’s hormonal and reproductive system health conditions were purchased for further analysis (Table 1).
Plant Names and Their Medicinal Parts That Are Used to Prepare Aromatic Waters for Women’s Hormonal and Reproductive Health Conditions.
Phytochemical Analysis
Volatile components of each beverage sample (500 mL) were extracted with 500 mL of petroleum ether using a glass liquid/liquid extractor system during 150 minutes. Chloroform was used for liquid/liquid extraction of ginger hydrosol. In this technique, the solvent vapor was transferred to the bottom of beverage container. The liquefied vapor in the beverage traveled to the top of the beverage container due to its lower density. Meanwhile, volatile components of the sample were transferred from the aqueous phase to the petroleum ether phase. In order to increase the concentration of volatile components in the organic phase, after 150 minutes the used beverage was replaced with fresh beverage and then extracted for another 150 minutes. The volume of each extracts was decreased to approximately 10 mL at 40°C and 60 rpm using a basic rotary evaporator. 15
Gas Chromatography–Mass Spectrometry
The concentrated and dehydrated extract of each ample beverage was injected to a gas chromatography–mass spectrometer for the analysis of respective volatile components. Agilent Technologies 7890 gas chromatograph with a mass detector (Agilent Technologies model 5975 C) was used in this study. The gas chromatograph was equipped with a HP-5MS capillary column (phenyl-methylsiloxan, 30 m, 0.25 mm id; Agilent Technologies 19091S-433 [60°C to 325/350°C]) and a mass spectrometer (Agilent Technologies 5975 C), which was operating in EI mode at 70 eV. The interface temperature was 280°C, and the mass range was 30 to 600 m/z. The oven was heated at a rate of 5°C/min from 60°C to 220°C and then it was held for 10 minutes at 220°C. Helium was used as the carrier gas with a flow rate of 1 mL/min. The components were identified by comparing the mass spectra and retention times with those of reference compounds, or with mass spectra in NIST or Willey libraries or in literature. 16,17
Results and Discussion
Hydrosols and Their Phytochemicals
The aromatic waters soft drinks that are used for women’s hormonal and reproductive health conditions are listed in Table 1. The data were prepared according to the information gathered via questionnaires (Tables 1 and 2).
Aromatic Waters’ Indications for Women’s Hormonal and Reproductive Health Conditions as Well as Their Other Indications.
This study was designed to investigate the aromatic waters that are used in Persian folk medicine, but some of these aromatic waters and their applications listed in this article have been mentioned also in some traditional Persian manuscript such as Qarabadin-e-salehi 18 and Qarabadin-e-kabir. 11 Although most current ethnopharmacological knowledge in Iran has been derived from historical Persian manuscripts, 19 it seems that some also have been arisen and accepted in recent years. This might be due to impact of new research on medicinal plants extracts on the knowledge of traditional healers as well as the companies that produce such products, although as far as we know there are not much research studies to provide evidence based data on the effects of aromatic waters or to elucidate their constituents.
In ethnomedical surveys, frequency of citation can reflect a kind of cultural importance of species, which may result in more accurate and more informants’ data obtained from questionnaires. 20 The frequency of citations of each hormonal and reproductive application for these beverages in all gathered questionnaires is shown in Figure 1. The higher frequency of citation can show the higher importance of an application for any of these aromatic waters. As seen in Figure 1, in all of the questionnaires (100%), fennel aromatic water was suggested to start menstruation, yarrow aromatic water to regulate menstruation, and Chehelgeyah aromatic water to treat dysmenorrhea. On the other hand, only a few informants believed that valerian aromatic water can regulate menstruation. As seen in Figure 1, most of introduced aromatic waters were believed to have indication to start menstruation. The second frequent cited application was regulation of menstruation.

Frequency of citations in questionnaires for women’s hormonal and reproductive health conditions.
In order to roughly investigate the popularity of these beverages, manufactories also were asked to rank them from 1 to 18 according to their mean of annual production over the past 3 years. Since these data were confidential for these manufactories, a ranking system was applied. The aromatic water with the lowest level of production was ranked 1. The manufactories ranking data represented as mean ± SD are shown in Figure 2. Among the aromatic waters that have indication for women’s hormonal and reproductive health conditions, yarrow, white horehound, Chehelgeyah, and fennel aromatic waters had higher annual production level during the past 3 years. This popularity might be because of their efficacy, the aromatic waters’ organoleptic properties such as taste and aroma, or even possible side effects during longer periods of consumptions. This might be also, due to their other applications rather than their effects on the reproductive system.

Ranking (1-18) of annual production level of aromatic waters in different manufactories over the past 3 years. Data are represented as mean ± SD.
The plants that are used to prepare these aromatic waters belong to 17 families. Apiaceae, Lamiaceae, and Asteraceae had a greater portion than others (Table 1).
Most of these aromatic waters are prepared from aerial parts of the plants. Different effects on women’s reproductive conditions including aphrodisiac, galactogogue, induction or regulation of menstruation, thinning vaginal discharge, cleansing reproductive system tracts, prevention of abortion, delivery induction, antitumor, treating infertility, and treating morning sickness were mentioned for these aromatic waters. Most of these beverages were believed to have hot nature.
Other indications rather than women’s hormonal and reproductive health were also mentioned for these beverages, which are summarized in Table 2. As mentioned in the introduction, aromatic waters’ aroma and compositions are considerably irrelevant to the pure volatile oil they were co-distilled with. As far as we know, the chemical constituents of most of these aromatic waters have not been investigated scientifically. This study determined constituents of these aromatic waters by gas chromatography–mass spectrometry after liquid-liquid extraction. As seen in Table 3, which shows the results of gas chromatography–mass spectrometry analysis, thymol is major or second major component except for dill, white horehound, willow, Moderr, and yarrow aromatic waters, whose major constituents are dill ether, menthol, phenethyl alcohol, linalool, or camphor. Carvacrol was also detected in all of these aromatic waters except for Taadol.
Aromatic Water Constituents Resulting From Gas Chromatography–Mass Spectrometry Analysis.
According to both hierarchical cluster analysis and K-means, all the aromatic waters that contain thymol as the major constituent make a cluster that includes some subcultures (Figure 3, Table 4). Lemon balm, Persian leek, Chehelgeyah, and ginger aromatic waters make a distinct subcluster. The reason behind the observed similarities between these aromatic waters based on clustering analysis was the presence of comparable amounts of thymol (41.45% to 64.74%) and carvacrol (26.20% to 30.49%) in these aromatic waters. The certain similarity of fennel, Taadol, and chamomile was also seen by hierarchical cluster analysis, which might be due to the presence of comparable amounts of thymol as the major constituent (42.20%, 44.98%, and 34.35%), carvone (14.53%, 15.84%, and 8.12%), and carvacrol (6.71%, 4.12%, and 4.65%) in these 3 aromatic waters.

Cluster analysis of aromatic waters’ constituents based on hierarchical cluster analysis. The aromatic waters are the following: 1, chamomile; 2, lemon balm; 3, yarrow; 4, Chehelgeyah; 5, white horehound; 6, stinging nettle; 7, parsley; 8, Moderr; 9, fennel; 10, dill; 11, valerian; 12, Taadol; 13, willow; 14, Persian leek; and 15, ginger.
Analysis of the Aromatic Waters’ Constituents Based on K-Means by SPSS Software (10 Epochs of Training).
In contrast to other aromatic waters, thymol was fond in zero or trace amount in willow and yarrow aromatic waters. Their major constituents are also completely different from others, and they were clustered at distinct groups.
Based on clustering methods applied in this study, although some similarities could be found, composition of white horehound, Moderr, willow, and yarrow aromatic waters revealed more differences than others. The main components of these aromatic waters were menthol (36.27%, white horehound), linalool (26.69%, Moderr), camphor (41.88, yarrow), and phenethyl alcohol (55.73%, willow).
Literature Survey
We could not find any reports on chemical constituents of aromatic waters of the mentioned plants in Table 1. Thus, it was not possible to compare the results, but the major components of the reported essential oils are summarized in Table 5.
Profile of Essential Oils Reported in Literature for the Plants Used to Prepare Aromatic Waters With Indications for Women’s Hormonal and Reproductive Health Conditions.
For most of these aromatic waters including lemon balm, stinging nettle, valerian, fennel, Persian leek, ginger, and white horehound, the major components in the aromatic waters and essential oils are completely different. Different allyl sulfides were reported as the major components of the Persian leek essential oils 21 but none of these components were detected in the aromatic waters in the present study. In the case of dill essential oil, the major components were reported to be phellandrene, limonene, myristicin, followed by dill ether. 22 In the present study, the major compound in dill aromatic water was dill ether (40.9%), followed by thymol and carvacrol. On the other hand, the major components of parsley leaf (myristicin) and willow (phenylethyl alcohol) were similar in aromatic waters and reported essentials but their amount as well as nonmajor constituents were different (Tables 3 and 5). As was expected, comparing the results of this study on chemical composition of the aromatic waters (Table 3) with the previous reports on the plants’ essential oils (Table 5) shows that there is a significant difference between aromatic waters and essential oil components. This difference may arise from different water solubility of the volatile components. It is possible that some of these volatile components were not entered in water during the preparation procedure of aromatic waters. Also, it can be concluded that it is essential to consider different pharmacological and biological properties of the aromatic waters due to different chemical compositions compared with the pure co-distilled essential oils.
Different biological activities of the plants used to prepare the identified aromatic waters on the reproductive system or sexual hormone conditions were investigated in the literature and summarized in Table 6. We could not find any report on hormonal activity or effects on reproductive system conditions for any of the aromatic waters. But, for some of these plants including fennel, dill, chamomile, and ginger, some clinical trial have been reported on their essential oils or different extracts. Of course, we cannot compare the observed effects of the essential oils or other extracts of these plants with their aromatic waters due to differences in constituents as well as their concentrations but these reports might strengthen the hypothesis of the beneficial effects for these aromatic waters on women’s reproductive and hormonal health conditions.
Literature Survey on Biological Activities of Plants Used in Preparing Aromatic Waters With Indications for Women’s Hormonal and Reproductive Health Conditions.
For some of these plants mentioned in Table 6, different aqueous, ethanol, and methanol extracts or plant powders were investigated and it is not clear that if the volatile components had a role in the observed effects. On the other hand, for some others listed in Table 6, the medicinal parts that were used in these studies are different from those that are used to prepare the aromatic waters in Persian ethnomedicine. For oriental plane we could not find any related report. The present investigation was not intended to evaluate the efficacy of these aromatic waters, but high consumption of these aromatic waters in Persian folk medicine and nutrition culture might be related to their efficacy.
Conclusion
This article introduced some aromatic waters that are used for women’s reproductive and hormonal health conditions in Persian folk medicine with different popularity and market values. Almost in all investigated aromatic waters the chemical composition was remarkably different from the essential oils of the plants that were used for their preparations. Clustering these aromatic waters using SPSS software revealed that despite the differences in the plants genus, family, and the medicinal parts of the plants that are used to prepare them, some similarity can be identified in their chemical compositions. In most cases thymol, carvacrol, and carvone were the major constituents and may have a role in their biological activities.
Scientific investigation of these aromatic waters may lead to the development of some functional beverages and soft drinks as a safe way of administration of essential oils or even new lead components or therapeutic agents.
Footnotes
Authors’ Note
This study was part of the PharmD thesis project of Mehdi Afifi.
Acknowledgments
The authors want to thank Nahal Shamaeezadeh (PharmD student at Shiraz University of Medical Sciences) for assistance in some parts of the extracting procedures.
Author Contributions
AH wrote the draft and contributed in guidance, data collection, and revisions of the final version of the article. HE and MA contributed to data collection, analyzing data, and revising the final version of the article.
Declaration of Conflicting Interests
The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.
Funding
The authors disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This study was funded by Shiraz University of Medical Sciences (Grant No. 92-01-70-7067).
Ethical Approval
This study was an experimental and laboratorial work and did not require ethical approval.
