Physicochemical evaluation and essential oil composition analysis of Hyssopus cuspidatus Boriss from Xinjiang, China

Background: It is reported that the plant Hyssopus cuspidatus Boriss from Xinjiang has great value. This article deals with the detailed pharmacognostic evaluation of the crude drug H. cuspidatus Boriss. Materials and Methods: The essential oil of H. cuspidatus Boriss from Xinjiang, China, was extracted by the method of hydrodistillation and the chemical composition of the essential oil was analyzed by gas chromatography-mass spectrometry (GC–MS). Results: The yield of essential oil based on the dry weight of the plant was 0.6%(w/w). Fifty compounds accounting for 99.42% of the total oil were identified. The major components were oxygenated terpenes (66.33%), monoterpenes (26.14%), oxygenated sesquiterpenes (1.25%), and octane (1.85%). Conclusion: Oxygenated terpenes were the main group of the compounds. The physicochemical parameters presented in this article may be proposed as parameters to establish the authenticity of H. cuspidatus Boriss and can possibly aid pharmacognostic and taxonomic species identification.


INTRODUCTION
It is commonly known that Hyssopus cuspidatus Boriss has various uses in the Xinjiang medicinal system, but there are few high-quality human trials researching these uses. It has been used traditionally as an antispasmodic for its antispasmodic action, [1] expectorant, emmenagogue (stimulates menstruation), stimulant, carminative (digestive aid), peripheral vasodilator, anti-inflammatory and anticatarrhal agent, tonic, and sweat-inducer.
H. cuspidatus Boriss and its oil are mainly used in the treatment of respiratory diseases. Hippocrates recommended using hyssop to treat bronchitis. Today, H. cuspidatus Boriss is used in the treatment of nasal congestion and mild irritations of the respiratory tract. The essential oil contains pinocamphone and isopinocamphone, which have neurotoxic effects. [2,3] H. cuspidatus Boriss has two native and one cultivated species in China, which is one of the aromatic endemic plants in the Xinjiang Province, of China. This plant, with the common local name of Shenxiangcao or Uygur's name zufa, has been of interest to Uygur traditional medicine, especially in Arletai. Infusion obtained from the aerial parts of H. cuspidatus Boriss is used traditionally for the following purposes: carminative and antispasmodic, in the treatment of cough, bronchitis, colds and chronic catarrh, and also for its tonic effects on the digestive, urinary, nervous, and bronchial systems. [4,5] The literature survey showed that the evaluation of the physicochemical properties and the essential oil composition of H. cuspidatus Boriss officinalis have previously been done, but not for H. cuspidatus Boriss.
In spite of the numerous medicinal uses attributed to this plant, pharmacognosy information about this plant has not been published. Hence, the present investigation is an attempt in this direction, including the determination of physicochemical constants, the preliminary phytochemical screening, physicochemical analysis, and essential oil composition analysis of the water extract of H. cuspidatus Boriss.

Plant materials and reagents
The aerial parts of H. cuspidatus Boriss were collected locally from the Arletai mountain of Xinjiang Province, China. Voucher specimens were deposited in the Traditional Chinese Medicine College Museum of Chinese herbal samples of Xinjiang Medical University.
Gas chromatography-mass spectrometry (GC-MS) analyses were carried out by using Shimadzu QP-2010 GC-MS system.

Preliminary phytochemical screening
The entire plant was ground to obtain a fine powder, which has been put into experiments. Preliminary phytochemical screening was carried out by using standard procedures according to the official method prescribed. [6] To detect the major chemical groups in H. cuspidatus Boriss by qualitative chemical tests, the entire plant extract was obtained by using petroleum ether, ethanol (95%), and water.

Physicochemical analysis
Physicochemical analyses to obtain the percentage of ash values and extractive values were performed according to the official methods prescribed in the book of extraction and abstraction of active ingredient from plant drug. [7] Fluorescence analysis was carried out according to the method of Chase and Pratt [8] and Kokoski et al. [9] Essential oil composition analysis The sample (100 g) was steam distilled with a Clevengertype apparatus for 6 h. The oil was collected and dried over anhydrous sodium sulfate, then stored at 4°C for analysis.
GC-MS analyses were carried out by using a Shimadzu QP-2010 GC-MS system, operated in the EI mode at 70 eV with scanning from 41 to 450 amu at 0.5 s, using a     The identification of the components was made by comparison of their retention time with respect to the n-alkane series (C 6 -C 22 ) internal standards. The mass spectra and relative retention indices (RI) were compared with those of commercial (NIST 05 and NIST 05 s). Area percentages were obtained from the TIC response without using an internal standard.

RESULTS AND DISCUSSION
Preliminary phytochemical screening performed on the various extracts disclosed the presence of terpenes in the petroleum ether, amino acid, flavanoids, terpenes, phenolics, tannins, carbohydrates, and organic acid in ethanol; amino acid, flavanoids, phenolics, and tannins, carbohydrates, and organic acid in water. The results are presented in Table 1.
The total ash value is 4.18% (w/w), acid-insoluble ash value is 0.28% (w/w), most of the total ash is soluble in acid, so acid-insoluble ash value is very low. Meanwhile, the water-soluble ash value is 1.56% (w/w), the watersoluble extractive is very high in this plant, extractive value is 11.47% (w/w). But ethanol-soluble extractive value and ether-soluble extractive value is low, 2.50% (w/w) and 1.79% (w/w), respectively. The results are presented in Tables 2 and 3.
The result of fluorescence analysis of the entire plant power of H. cuspidatus Boriss is presented in Table 4, the fluorescence in day light, UV light (254 nm), and UV light (365 nm) are different from each other when different chemical reagents are added. It indicates that there are various chemical groups existing in the plant.
The air-dried aerial parts of H. cuspidatus Boriss were reduced to coarse powder and the oil was isolated by hydrodistillation using a Clevenger-type apparatus for 6 h. The oil was subsequently dried over anhydrous sodium sulfate. The volatile light orange yellow oil (0.6% w/w) was analyzed by GC-MS [ Figure 1]. The composition of the essential oil is listed in Table 5 along with the RI of the identified compounds, where all constituents are arranged in such a way that their elution was on the DB-5 Retention indices relative to C 6 -C 22 n-alkanes on the DB-5 column-preliminary phytochemical screening.