2.3.3. Statistical Analysis

All mineral data are reported as the mean and standard deviation of three independent determinations. With regards to the selected heavy metals (Cd, Pb, As, and Hg), their contents were statistically compared with the reference limit values established by the European Commission and WHO by means of Student's t test. The statistical analysis has been performed using the Past software (Version 4.09) [37].

#### **3. Results and Discussion**

The safety aspects that were explored throughout the study were: (i) microbiological aspects; and (ii) mineral profiles.

## *3.1. Microbiological Analysis*

The microbiological analysis of 13 dried spices, labelled with letters from A to O, is reported in Table 2.

**Table 2.** Bacterial contamination (CFU g<sup>−</sup>1) of dried spices from India (**a**) and Iran, Indonesia, and Vietnam (**b**).



a Reference values for microbiological safety and quality were based on regulations from the European Union and other international guidelines [8,38,39]. b CFU 25 g<sup>−</sup>1. \* Values are higher than the reference values. ND, not determined. A, black cumin seeds; B, Iranian Tokhme Sharbati; C, clove buds; D, Shahjeera; E, Abbaszadeh saffron; F, organic fenugreek; G, whole black pepper; H, cinnamon; I, Abthul Ahmar (Asario); L, Ajwan seeds; M, whole coriander seeds; N, black sesame seeds; O, Sabja seeds.

According to the Commission Regulation (EC) No. 2073/2005 and its amendment Regulation (EC) No. 1441/2007, which sets legal microbiological criteria for several food products, these spices could be defined safe products [8,38,39].

With regards to the indicator microorganisms, which are used to provide simple, reliable, and rapid information about processing failures, post-processing contamination from the environment, the general level of hygiene, and the presence or absence of foodborne pathogens to monitor the chain of food production, low bacterial loads were detected in saffron, fenugreek, black pepper, cinnamon, cress sprouting seed, thymol seed, coriander, and chia seed [40].

The total coliform and Enterobacteriaceae amounts in black cumin were 7 × 10<sup>3</sup> CFU g<sup>−</sup><sup>1</sup> and 1.4 × 10<sup>4</sup> CFU g<sup>−</sup>1, respectively, whereas in Iranian chia seeds it was 1.6 × 10<sup>3</sup> CFU g<sup>−</sup><sup>1</sup> and 6.6 × 10<sup>2</sup> CFU g<sup>−</sup>1, respectively. Caraway presented a mesophilic count of 1.7 × 10<sup>6</sup> CFU g<sup>−</sup>1, and an amount of total coliform and Enterobacteriaceae of 2.4 × 10<sup>4</sup> CFU g<sup>−</sup><sup>1</sup> and 9 × 10<sup>4</sup> CFU g<sup>−</sup>1, respectively. Moreover, Enterobacteriaceae were also detected in clove buds (2 × 10 CFU g<sup>−</sup>1) and black sesame seeds (4.4 × 10<sup>3</sup> CFU g<sup>−</sup>1).

These contaminations may be related to the environment, inadequate hygienic handling, unsanitary conditions, and others, and occurs in samples purchased in street markets. Indeed, the adoption of good hygiene practices in all the involved steps from land growing, harvesting, and processing can be useful to reduce the health risks of spice consumption.
