BİYOLOJİK AKTİVİTELERİ

Fig. 1. Hibiscus sabdariffa L. plant and its biological potentials validated so far.

It was a fact that, HS has been used to treat cardiac and nerve conditions, induce diuresis, and
induce lactation, among other conditions in folklore. Logical examinations have additionally
exhibited the antihypertensive, antidiabetic, against weight and hostile to hyperlipidaemic
properties of HS. These organic exercises are believed to be because of the battery of
phytochemicals in HS that have solid cell reinforcement movement and that hinder α-
amylase, α-glucosidase, angiotensin changing over catalyst, calcium channel blockage and
direct vasorelaxant impacts. A portion of the phytochemicals that are believed to be liable for
these natural impacts incorporate anthocyanins, flavonoids and natural acids. There is anyway
the requirement for more hearty investigates including controlled clinical preliminaries to
approve these organic exercises with the end goal of carrying the advantages nearer to the
bedside.
Hibiscus sabdariffa contain following various biological activities:
Anorexia; Atherosclerosis ; Bacteria ; Biliousness ; Cancer ; Carbuncle ; Cardiopathy ; Catarrh ;
Conjunctivosis ; Constipation ; Chill ; Circulosis ; Cold ; Constipation ; Cough ; Cramp ; Debility ;
Dyspepsia ; Dysuria ; Enterosis ; Fever ; Gastrosis ; Hangover ; Heart ; Herpes zoster ; High Blood
Pressure ; Infection ; Inflammation ; Insomnia ; Intoxication.

DISTRUBITON

Origin of Hibiscus sabdariffa

There is a big argument about the origin of Roselle among
different scholars. Cobley suggested Roselle is a native plant
of West Africa and from there it was carried to other parts of
the world such as Asia and America, whereas in others opinion,
Roselle was originated from India and Saudi Arabia
PHYTOCHEMISTRY of HIBISCUS SABDARIFFA
Proximate constituents and nutrient composition of Hibiscus sabdariffa

Even though, the uses of different parts of Roselle are many and varied both in food and in traditional
medicine, all parts of Roselle including seeds, leaves, fruits and roots are used as a food in different
parts of the world. Fleshy red calyxes of Roselle are commonly used for the production of soft drinks
and tonic without alcohol like wine, juice, jam, jelly, syrupand also dried and brewed into tea and
spice. These are rich in carotene, riboflavin, anthocyanins, ascorbic acid, niacin, calcium, iron and
vitamin C. The young leaves and tender stems of Roselle are consumed raw as green vegetable. The
Roselle seeds are good source of protein, fat, total sugars and are widely used in the diet in many
African countries

PHYTOCHEMICALS OF HIBISCUS SABDARIFFA

Bioactive components
The main constituent of roselle in context of therapeutic importance are a polysaccharide, organic acid
and flavonoids mainly anthocyanins [50–53]. The extracts of dried calyces have been known to
contain chemical constituents such as organic acids (citric acid, ascorbic acid, maleic acid, hibsicic
acid, oxalic acid, tartaric acid) besides, phytosterols, polyphenols, anthocyanins and other water-
soluble antioxidants [10,28]. The organic acids together with bioactive components have free radical
scavenging activity [36]. The beneficial health effect is mainly attributed to these bioactive molecules.
Table 1 shows the polyphenolic fraction (bioactive compounds) present in the extracts of Hibiscus
sabdariffa as reported by different research groups.

5.1. Organic acid

Roselle extracts contain a high percentage of organic acids including malic acid and citric acid (13%
on the dry weight basis) in the calyces as reported by Salma and Ibrahim [56]. Besides, Abou-Arab et
al. [37] reported that the calyces also contain ascorbic acid (140.13 mg/100 g). Earlier data also show
that the calyces are rich in ascorbic acid [47,56,57]. Some authors have reported high concentration of
organic acids such as oxalic acid, succinic acid, tartaric acid; malic acid, citric acid with the last of
these predominating [1]. Recently Jabeur et al. [40] have reported oxalic acid, shikimic and fumaric
acids as the main organic acid with malic acid (9.10 g/100 g) to be the most abundant acid in the
calyces of roselle.

5.2. Polyphenols and flavonoids

The origin of many therapeutic substances is due to the secondary metabolisms in the plant. Roselle
calyces are an interesting source of potential bioactive molecules with antioxidant,
hypocholesterolemic antihypertensive, antimicrobial, anti-inflammatory, antidiabetic and
anticarcinogenic activities. Many scientific investigations have revealed that the calyces of roselle are
rich in polyphenols and flavonoids that enhanced the nutritive value of roselle as these compounds are
correlated with their antioxidant property. The phenolic content in the plant consists mainly of
anthocyanins like delphinidin-3-glucoside, sambubioside, and cyanidine-3- sambubioside [40,54] and
other flavonoids like gossypetine, hibiscetin and their respective glycosides; protocatechuic acid,
eugenol, and sterol like β-sitoesterol and ergoesterol [1,10,58]. The colourful anthocyanins are the
members of the flavonoids group of phytochemicals. The anthocyanins are flavylium or 2-
phenylbenzopyrylium cation with hydroxyl and methoxyl group present at position R1 and R2 of the
basic structure shown in Fig. 3 [59]. Anthocyanins molecules are susceptible to degradation. Their
stability depends on pH, temperature, the presence of enzyme, light, and structure, the presence of
other flavonoids, phenolic acids and metals [60]
5.2.1. Anthocyanins of Hibiscus sabdariffa L
Ali et al. [1] and Gradinaru et al. [61] have reported two major anthocyanins compounds,
(delphinidine-3-sambubioside and cyanidine-3-sambubioside) and two minor compounds
(delphinidine-3-glucoside and cyanidine-3-glucoside) present in the calyces of roselle. Earlier Pouget
et al. [62] and Bridle and Timberlake [63], have reported the similar findings. According to a study
conducted by Tsai et al. [64], have reported that 85% of anthocyanins is delphinidine-3- sambubioside
and is the major source of antioxidant capacity of roselle extract. In another study by Aurelio et al.
[35] reported that the extracts of roselle calyces are rich in anthocyanins like delphinidine-3-glucoside,
sambubioside and cyanidin-3-sambubioside contributing to their antioxidant properties. The
researchers have mainly used an aqueous or organic solvent to extract the polyphenols and
anthocyanins from the roselle calyces. The different extraction techniques and different varieties of
Hibiscus sabdariffa used in various studies make it difficult to compare. Luvonga et al. [36] reported
total phenolic content to be 6.06 mg/g in roselle extract. Dried roselle contained total anthocyanins as
cyanidine 3-glucoside 622.91 mg/100 g and 37.42 mg/g total phenolic content in dry weight sample
[37]. Recently Jabeur et al. [40] in their study identified delphinedine-3-o-sambubioside, delphinidine-
3-o glucoside and cyanidine-3-o sambubioside in the concentration of (7.03 mg/g), (1.54 mg/g) and
(4.40 mg/g) respectively
4.PHARMACOLOGICAL EFFECT and CLINICAL TRIALS
Anti-oxidant effect:
Protective property of a compound to inhibit the oxidative mechanisms by scavenging reactive
oxygen and free radicals is known as antioxidative activity. It protects lining organelles from
premature cell damage and reduces ageing. A large number of invitro and invivo studies have shown
that Roselle calyxes contain potent antioxidant. According to Augustine [38], both the whole aqueous
and anthocyanin-rich extracts of Roselle are effective antioxidant. Studies have also highlighted that
polyphenolic acid, flavonoids and anthocyanins which are found in Roselle are potent antioxidants
[39].

. Antihyperlipidemic
Various studies have shown the hypolipidemic efficacy of Hibiscus sabdariffa extract (HSE)
suggesting the possibility of roselle as an antiobesity agent. Obesity is characterized by a gain in the
body weight due to excessive accumulation of body fat. High-fat diet and energy imbalance, results in
obesity which is associated with various metabolic syndromes like cardiovascular risk factors
(hyperlipidemia and hypertension), fatty liver and insulin resistance [75,76]. These metabolic
disorders are characterized by early cellular events and dysregulation of normal cellular homeostasis.
Polyphenols derived from Hibiscus sabdariffa has pleiotropic character and could become auxiliary in
this pathology as they have multi-targeted effects on obesity affecting human health and this multi-
targeted mechanism involves regulation of energy metabolism, oxidative stress and inflammatory
pathways, transcription factors, hormones and peptides, digestive enzymes, as well as epigenetic
modifications [8]. There are evidence from animal studies and human clinical trials that HSE reduced
hyperlipidemia. In a study it was reported, that when hypercholesterolemic rats were fed with a dose
of 5–10 % of HSE for 9 weeks lowered the different lipid fraction in plasma, heart, brain, kidney and
liver [77]. Similarly, in another study, it was reported that when rabbits fed with cholesterol for 10
weeks and then treated with 0.5 or 1% HSE showed that the levels of, total cholesterol, triglyceride
and low-density lipoprotein (LDL) in the serum were reduced [33]

Antidiabetic
Evidences are accumulating on diabetes mellitus control potential of H. sabdariffa. Peng et al. [24]
analyzed the composition of H. sabdariffa polyphenolic extract and used a type 2 diabetic rat model to
test its protective effect. Treatment with the phenolic-rich extract reduced hyperglycemia and
hyperinsulinemia, with maximum efficacy at 200 mg/kg. The extract decreased serum triacylglycerol,
cholesterol, and the ratio of low-density lipoprotein/high-density lipoprotein. The extract significantly
reduced the abnormal elevation in plasma advanced glycation end (AGE) formation by inhibiting the
expression of their receptors. Adisakwattana et al. [25] investigated plants, including H. sabdariffa, for
intestinal -glucosidase and pancreatic -amylase inhibition. When combined with chrysanthemum,
butterfly pea and mulberry extract, it exerted additive intestinal maltase inhibition. On the other hand,
its combination with chrysanthemum and mulberry caused synergistic inhibition of pancreatic -
amylase. Ademiluyi and Oboh [26] also investigated the inhibitory effect of aqueous extracts of two
varieties (red and white) of H. sabdariffa calyces on -amylase and -glucosidase. The red variety
exhibited higher -glucosidase inhibitory activity, whereas the white variety exhibited higher -amylase
inhibitory activity. The -glucosidase inhibitory activities of both calyces were higher than that of their
-amylase. The red variety possessed strong antioxidant capacity due to high phenolic content.
Wisetmuen et al. [27] investigated the mechanism underlying the antihyperglycemic activity of its
ethanolic extract in diabetic rats. When administered at a dose of 1 g/kg daily for 6 weeks, the extract
significantly decreased the blood glucose level and increased the basal insulin level. Histological
examination showed that the extract increases the number of islets of Langerhans in diabetic rats.
Collectively, the inhibition of intestinal -glucosidase and pancreatic -amylase activities, combined with
antioxidant abundance renders H. sabdariffa as a promising adjunct therapy for diabetes.

Antihypertensive
Hypertension is a chronic medical condition of the heart in which the arterial blood pressure is raised.
This is a common health problem in both developed and developing countries. Traditionally Hibiscus
sabdariffa (HS) is used as the antihypertensive agent. The ancient use of HS is related to
cardiovascular diseases. Several scientific investigation reports have provided evidence that Hibiscus
sabdariffa extract (HSE) significantly reduces blood pressure in both animal and human model.Haji
faraji and Haji Tarkhani [87] have evaluated the effect of HS tea on hypertension. The first single trial
was studied in 54 patients with moderate hypertension which revealed a reduction in (systolic by
11.2% and diastolic by 10.7%) blood pressure in 12 days after beginning of the treatment and when
the treatment was stopped, after three days the systolic and diastolic blood pressure was elevated by
5.6% and 6.2% respectively. In a randomized trial of 75 patients, Herrera-Arellano et al. [88] have
shown that HS aqueous extract effectively reduced the blood pressure by more than 10% and was safe
when compared with 25 mg of captopril. In another study again in Herrera-Arellano et al. [89]
highlighted the effect of standardized Hibiscus sabdariffa extract in a randomized, double-blind,
lisinopril-controlled clinical trial involving 171 hypertensive patients for 4 weeks. HS extract lowered
the blood pressure (BP) but the magnitude of the effect was lower when compared with the patient
treated with 10 mg lisinopril concluding that Hs exerted important antihypertensive effect with a wide
margin of tolerability and safety.

hepatoprotective

Protective effects of dried flower extracts against oxidative stress in rat primary hepatocytes were
demonstrated31. Protocatechuic acid, a simple phenolic compound isolated from H. sabdariffa showed
protective effects against cytotoxicity and genotoxicity of hepatocytes induced by t-BHP. One of
mechanisms may be associated with its property of scavenging free radicals32. The extract of its petals
protected rats against cadmium induced liver, prostate and testis lipoperoxidation33. The extract offers
hepatoprotection by influencing the levels of lipid peroxidation products and liver marker enzymes in
experimental hyperammonemia and this could be due to the free radical scavenging property of natural
antioxidants present in the plant34. The protective effect of aqueous extract and anthocyanins on
paracetamolinduced hepatoxicity in rats has also been reported35. Aqueous-ethanol (1:1) extract of the
calyx showed a significant decrease in the level of lipid peroxidation in carbon tetrachloride induced
liver damage36. However, a study showed that prolonged usage of aqueous-methanol extract of the
calyces could cause liver injury37.

antibacterial
The antibacterial activity of methanol extract of H. sabdariffa calyces was evaluated by disc diffusion
method as mentioned in Abdallah and Al-Harbi [15]; 100 μl from each working bacterial cultures were
mixed with 20 ml warm autoclaved Mueller-Hinton agar (Watin-Biolife, KSA) in glass bottles size 50
ml, tighten and mixed well and then poured directly into 90 mm sterile plastic disposable plates (Jalil
Medicals) and left to solidify at room temperature. 6 mm discs were previously prepared using
Whatman No.1 filter paper, sterilized and saturated with 300 mg/ml of the methanol extract of H.
sabdariffa calyces, the pre-experimental test showed that the blank disk (6 mm) could carry 20 μl of
the extract at a concentration of 500 mg/ml, which equivalent to 10 mg/disc. Then, the wet, saturated
disks were directly loaded on the cultured Mueller-Hinton Agar plate in aseptic conditions. On the
same plate, 6 mm antibiotic discs; penicillin G 10 units and gentamicin 10 μg (Oxoid) were also
loaded, which served as positive controls. Cultured plates with extract and antibiotic discs were
incubated for about 24 h at 37°C. The mean zone of inhibition of two replicated disks on the same
plate was taken in millimeter (mm) using a ruler, 6 mm zone diameter considered as no inhibition.
immunomodulation
The concept that immune responses are modulated to alleviate diseases has existed in many forms of
traditional medicine beliefs, with plants being used in such systems to promote health and to maintain
the body’s resistance against infections by potentiating immunity. Some of these plants are specifically
stimulatory or suppressive, and normalize or modulate pathophysiological processes, and are thereby
termed ‘immunomodulatory’.

to some extent the extracts, and to a large extent, two fractions of the plant possess the ability to
stimulate the immune system in vivo. The activity may be as a result of interplay between the
production of interleukin 10, inhibition of tumor necrosis factor-alpha and the effect of B-cells
responsible for antibody production. One of the fractions showed good possibilities of being
developed into a drug entity that may be used to stimulate immunity as an adjunct to therapy in
immunosuppressed disease conditions. However, the extract, as it is taken in humans as a beverage
may be of benefit in enhancing immunity. Further studies need to be done to evaluate the exact
mechanism of action.

diuretic

The aqueous extract from this plant and the fractions obtained with solvents of different polarities
were administered to adrenalectomized rats, and the diuretic effect was measured in the presence of
deoxycorticosterone acetate (aldosterone analog).The effect on renal filtration was also evaluated in an
in situ kidney model, and finally, the effect of diuretic active extracts on gene expression of the alpha
subunit from the transporter (αENaC) of renal epithelial cell was quantified. The subsequent results
were obtained: The aqueous extract of Hibiscus sabdariffa presented the following chemical
composition, 32.4 mg/g delphinidin-3-O-sambubioside, 11.5 mg/g cyanidin-3-O sambubioside, 11.5
mg/g quercetin, and chlorogenic acid 2.7 mg/g. The concentration of anthocyanins was diminished
until disappearance due to decrease of the polarity of the solvents used in the extraction process, in
contrast to the flavonoids and chlorogenic acid, which had their concentration increased. The diuretic
effect caused by adrenalectomy in rats was reversed by deoxycorticosterone acetate activity. However,
the effect of deoxycorticosterone acetate was antagonized by spironolactone, the aqueous extract of
Hibiscus sabdariffa, and the acetonitrile :methanol 5 : 5 mixture extract, administered orally. A similar
effect was observed on renal filtration obtained from the isolated kidney model. is that the diuretic,
natriuretic, and potassium sparing effects of Hibiscus sabdariffa are due in part to the modulation of
aldosterone activity by the presence in the extract of this plant of compounds potentially responsible
for this modulation, as anthocyanins,flavonoids, and chlorogenic acid.

The compound presents in Hibiscus sabdariffa as quercetin had effect on the vascular
endothelium causing oxide nitric release, increasing renal vasorelaxation by increasing
kidney filtration. Therefore, the diuretic effect of Hibiscus sabdariffa may be mediated by
nitric oxide release. The compound presents in Hibiscus sabdariffa as quercetin had effect on the
vascular endothelium causing oxide nitric release, increasing renal vasorelaxation by increasing
kidney filtration. Therefore, the diuretic effect of Hibiscus sabdariffa may be mediated by nitric oxide
release.
Treatment of anemia
Hibiscus sabdariffa (Hs) was the most cited species against anaemia being rich in iron and
ascorbic acid. This ascorbic acid helps in the absorption of non-heme iron which justifies their
potential as anti anaemic in ethnomedicine. Pharmacologically, Hibiscus sabdariffa (HS)
extract was tested in both the animal and human models results, showed the elevated
hematocrit and haemoglobin [66]. Adigun et al. [105] evaluated the effect aqueous extract of
Hibiscus sabdariffa on haematological parameters like haemoglobin, haematocrit and total
and differential white blood cells count recommending a dosage of 200–400mg/kg to be
beneficial. Doses of 200–1000mg/kg body weight were given orally to rats up to 14 days.
After 14 days significant elevation were observed in haematocrit and haemoglobin in the
group of animals given doses of 200–400mg per kg. However higher doses showed a
significant reduction in the haematocrit level but not in the haemoglobin. suggesting that
administration of aqueous extract of Hibiscus sabdariffa is beneficial to the haematopoietic
system. Anaemia is an iron deficiency disease having low red blood cells (RBC) in the
blood. The RBC carries and delivers the oxygen to the body cells. The anti-anaemic activity
of roselle extract might be because of its very low pH and high concentration of ascorbic acid
that has increased the mineral bioavailability [2]. Research on anti-anaemic activity of roselle
extract is limited and better quality research is required, with bigger sample size and good
study design, to investigate the possible potential of roselle in the treatment of anaemia and
their mechanism of action which is an emerging area.

anticancer effect
Anthocyanins can cause cancer cell apoptosis, especially in HL-60 cells46. Anti-oxidative activity of
anthocyanins was evaluated by their effects on LDL oxidation in cell free system and anti-apoptotic
abilities in RAW264.7 cells47. The study showed that anthocyanins of this plant may be used to inhibit
LDL oxidation and oxLDL-mediated macrophage apoptosis, serving as a chemopreventive agent.
Inhibitory effect of protocatechuic acid on tumour promotion in mouse skin demonstrated that
protocatechuic acid possesses potential as a cancer chemopreventive agent against tumour
promotion48.

8. The dosage of the extract

It is difficult to elucidate dosing recommendation because of the heterogeneity in the studies
related to the bioactivity of the poly phenoliccontents of Hibiscus sabdariffa.Different
methods of extraction and products are used in different studies.The doses that are used in
each of the positive studies are gives.

Doses of the extract of Hibiscus sabdariffa with biological activity.

TOXICIITY
in recent years the effect of Hibiscus extracts on reproduction and development in rats has been
subject to in vivo studies by several authors. In female rats, addition of hibiscus extracts to drinking
water resulted in a dose-dependent reduction of fluid and feed intake. The effects observed in the pups
included increased weight gain and delayed onset of puberty. As the authors rightly state, these effects
have likely been caused by the reduced fluid and feed intake in accordance with earlier publications.
Rather than representing specific toxicologic properties of hibiscus, the decreased fluid and feed intake
and subsequent adverse effects observed in this study may be the result of the animals dislike for the
flavour of hibiscus (Iyare & Adegoke, 2008a,b; Iyare et al., 2010).

The plant extracts are characterized by a very low degree of toxicity. The LD50 of H. sabdariffa calyx
extract in rats wasfound to be above 5000 mg/kg (Onyenekwe et al., 1999),suggesting that the extract
is virtually non-toxic. Inspontaneously hypertensive rats, treatment with the extract at doses of 500–
1000 mg/kg decreased blood pressure, and also significantly decreased serum creatinine, cholesterol
and glucose levels, but significantly increased the serum content of uric acid.

The effect of roselle calyx consumption on kidney function, serum creatinine and uric acid levels in
hypertensive and normotensive conditions was measured. The serum creatinine level in the
hypertensive control group was significantly higher than the corresponding value in normotensive rats.