Article Microbial, chemical composition evaluation and development of a technological process for the production of compound spices in Nigeria Olalekan David Adeniyi*, A S Abdulkareem, B M Idris and J O Odigure Chemical Engineering Department Federal University of Technology, Minna, Nigeria *Correspondent author, E-mail : [email protected] volatile oils, which contains terpene, Abstract aldehydes, ketones and alcohol of various phenol based compounds. Spices are vegetable products derived from fruits, seeds, roots Most spices and condiments do not and tree barks. They are important mainly as additives to food because have any reasonable bacteriostatic of the presence of essential oil having anti-microbial and fungicidal effect on the concentration of the properties. These properties have been used to some degree as the basis food material they are used on.1 The for food preservation and as medicinal products for certain types of United States, Food and Drug diseases in Nigeria. This paper takes a look at the microbial load and Administration (FDA) specifies that chemical compositions of some of the compound spices found in Nigeria. paprika, saffron and turmeric are Results reveal that some compound spices are highly contaminated with colours and they must be labelled microorganisms. Isolates from sample B were identified as Salmonella, with common names on food Bacillus and Escherichia coli bacteria while those from sample A labels.1, 2 contained Staphylococcus aureus and Streptococcus. The fungi isolates Normally high were Aspergillus niger, A. flavus and Microsporum caris. The bio-load concentrations of plant extracts are of the spices ranged from 1.64 105 to 2.09 107 cells per gram of spice. × × required before anti-microbial Salmonella species and Bacillus sp. were the most common properties become apparent. It is microorganisms. It is recommended that higher sanitary conditions be well documented that vegetables employed during the processing, handling and storing of the spices. This like, onions, garlic and horse-radish paper proposes a technological process route for the production of more provide anti-microbial extracts.3,4 hygienic spices. For centuries it has been known that certain foods especially spices Introduction seasonings and imparting aroma to possess naturally occurring 1 food. Spices and condiments are preservative qualities but only The definition of spices, usually grown in tropical or semi recently has the technology been herbs and condiments is still the tropical climates and comprise of developed to extract, quantify and focus of much debate over the years. one portion of the plant. Spices are identify these substances. The International Organization for the whole or ground seeds, fruits, Spices vary in their Standardization (ISO) defines spices bark or roots of a plant. effectiveness depending upon the and condiments as the natural Spices have characteristic source, freshness and method of vegetative products or mixture odours or flavours. The chemicals processing and storage. The thereof without extraneous matter responsible for their distinctive inhibitory effect of spices differs that are used for flavourings, tastes and smell are essential oils or with the kind of spice and the 314 Natural Product Radiance Vol 2(6) November-December 2003 Article microorganism being tested for. Pseudomonas aeruginosa were the Methodology Mustard and the volatile oil of most resistant except towards thyme mustard have varying effectiveness oil. Antibacterial and antifungal The material used for the against Saccharomyces cerevisiae effectiveness of the compounds of experiment include spice product but are not effective against most the spice Aframomum melegueta such as garlic (Allium sativum bacteria as cinnamon and cloves.1,5,6 have been carried out by Oloke et Linn.), ginger root (Zingiber Cinnamon and cloves contain al7 and the volatile oil showed officinale Rosc.), allspice [Pimenta cinnamic aldehydes, which makes considerable bactericidal activities dioica (Linn.) Merrill], mansoro, them more bacteriostatic than against Escherichia coli, cloves [Syzygium aromaticum mustard. Thyme, bay leaves, Pseudomonas, etc. and fungicidal (Linn.) Merrill & Perry], gyada marjoram saucy, rosemary, black activities against Candida albicans, muyan, thyme (Thymus vulgaris pepper and others have weak Trichophyton mentagrophytes, etc. Linn.), crayfish, alligator pepper inhibitory power against most These properties have been [Aframomum melegueta (Rosc.) organism. used to some degree as the basis for K. Schum.], white pepper, African Several studies have been food preservation. However, certain hot pepper and locust beans conducted using different spices to spices are known to contain large (Ceratonia siliqua Linn.). The inhibit the growth of pathogenic number of microorganism despite spices were gathered from the local organism. Kivanc and Akgul6 tested their apparent anti-microbial effect. farm and market, cleaned and sun the bacteriostatic and bactericidal Some of these organisms and dried at temperature of 45°C activities of twenty-two essential chemical content may cause maximum, until a constant dried oils from Turkish spices and citrus spoilage or become pathogenic to weight was attained. It was then against Aerobacter aerogenes, man when introduced to food.10 passed through magnetic separator Bacillus subtilis, etc. The results There is little information available to remove the foreign material showed that the essential oils tested on the microflora, volatile oil, manually, sieved to remove the dirt, varied in their antibacterial activity. production techniques and quality of insects and ground to fine particles Anise, celery, coriander and sage local Nigerian spices. This paper of size 0.05 µm. Two samples (A & were inactive or had little activity presents results of some of the B) as proposed by Djmen11 for while corn, mint, cumin, lemon peel analyses carried out on the local mixing by weight were used; the and ziziphora were active against all spices and proposes a technological ratios are given in Table 1. tested bacteria to a variable extent. process for the production of more Staphylococcus aureus and hygienic spices. Table 1 : Mixing ratio by weight (wt.) for spices mixture (samples) Sample A Sample B 10 % wt. Garlic 5%wt.Gyanda muyan 15 % wt. Garlic 10 % wt. Thyme 10 % wt. Ginger 10 % wt. Thyme 15 % wt. Ginger 10 % wt. Crayfish 5 % wt. Allspice 5 % wt. Crayfish 5 % wt. Allspice 10% wt.White pepper 10%wt. African 10% wt. Alligator 15%wt. African 10% wt. Alligator hot pepper pepper hot pepper pepper 5 % wt. Mansoro 20% wt.White pepper 5 % wt. Mansoro 10 % wt. Cloves 5 % wt. Cloves Natural Product Radiance Vol 2(6) November-December 2003 315 Article estimating the difference in fresh expelling the contents of the pipette. and dry weight. This pipette was also discarded and the second dilution tube was labell- Determination of protein ed 10-2. Taking fresh sterile pipettes and carbohydrate in each case other specimen were -3 -4 -5 Protein content was prepared for 10 , 10 and 10 . measured by Kjedahl method and Plate preparation procedure the total carbohydrate was Taking a fresh sterile pipette determined by using estimation the content of the final dilution tube method.8 If the total of protein, lipid (10-3) was mixed by sucking up and acid content was subtracted from down five times. One ml of the organic matter, the remainder dilution was withdrawn by touching accounts for carbohydrate and the tip of the pipette against the side nucleic acid. Thus, % carbohydrate Ceratonia siliqua of the tube to remove excesses = 100 - (% moisture + % ash + % adhering to the outside and the protein + % lipid + % fibre) contents was transferred to a sterile contents. petri dish. Allowing about 3 minutes Microbiological analysis of to elapse the tip of the pipette was samples A and B touched against the dish away from the previous inoculum and the Dilute Solution Preparation remaining drops gently blown away. Procedure This same pipette was used to Syzygium aromaticum A sterile 1 ml blowout transfer 1 ml from the 10-2 dilution pipette was held vertically and the to a sterile petri dish but before pipette tip was introduced to the taking the samples it was raised and surface of samples A and B and lowered for about three times in sucked up and down five times to order to rinse the sides of the pipette the 1 ml mark. One ml of samples and also to give the dilution a final A and B was withdrawn. The pipette mixing. The same procedures were content was transferred to the first repeated for the 10-1 dilution. tube of dilution solution series with To each plate 15 to 20 ml of the tip touching the side above the molten compound spices agar level of the diluents. This pipette (bacterial) and PDA medium at 45°C was then discarded and the first was added and immediately the dilution tube was labelled 10-1. medium and inoculum was mixed Taking a fresh sterile pipette, by a combination of to and fro and Pimenta dioica the content of the first dilution tube shaking for about 5-10 seconds. The Determination of moisture was mixed by sucking up and down plates were allowed to set, then to the 1 ml mark five times. Then inverted and incubated and ash content 1ml of the first dilution was at the appropriate temperature of The moisture and ash withdrawn and transferred to a 25-30°C for fungi and 37°C for contents were determined by second tube of sterile dilution bacteria.1, 8, 9, 12, 13, 14 316 Natural Product Radiance Vol 2(6) November-December 2003 Article Fungi analysis Preparation of nutrient agar thoroughly cooked. The suspension was filtered through cheese cloth Twenty-eight grams of and water added to make up to 1 ml. Preparation of compound spices compound spices agar powder was Dry ingredient was also added and broth weighed and dissolved in 100 ml of agar was dissolved with heat. distilled water. It was boiled, stirred Sterilization was conducted in an Thirteen grams of and autoclaved at 121°C for 15 autoclave at 121°C for 15 minutes.